Molecular Insight and Antioxidative Therapeutic Potentials of Plant-Derived Compounds in Breast Cancer Treatment

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

This review manuscript provides a comprehensive overview of the potential of plant-derived compounds (phytochemicals) in breast cancer treatment, with a specific focus on their molecular mechanisms and antioxidative properties. The authors cover a broad range of topics, including the historical context of phytotherapy, key oncogenic pathways in breast cancer, specific phytochemical classes and their actions, the role of antioxidants, and challenges in clinical translation. The inclusion of detailed figures illustrating molecular pathways (Fig. 2) and specific examples of phytochemical actions with experimental data (Figs. 3, 4, and 5), as well as informative tables summarizing compound activities (Tables 1 and 2), are definite strengths.

The manuscript is ambitious in its scope and generally well-structured. However, while "antioxidative" is in the title and the journal's focus, the direct and primary link between the antioxidant properties of many discussed compounds and their specific anticancer molecular mechanisms could be more explicitly and consistently elaborated throughout the text. In some sections, the antioxidant aspect feels secondary to the broader anticancer effects. The manuscript would benefit from a clearer delineation of when antioxidant activity is the primary mechanism versus a contributory one and how this specifically influences the discussed molecular pathways. Further polishing of the language and ensuring consistent focus on the "antioxidative therapeutic potentials" will enhance its impact.

Major Issues:

1. Elaboration on the "Antioxidative" Link:
   • While Section 3 directly addresses "Therapeutic Potentials of Antioxidants," the subsequent sections (e.g., Section 5 on "Antioxidative Phytochemicals as Molecular Modulators") discuss various phytochemicals and their effects on signalling pathways, but thespecific role of their antioxidant properties in modulating these pathways is not always clearly articulated. For example, when discussing resveratrol's effect on YAP/TAZ (Figure 3) or hesperidin's effects (Figure 4), the text describes pathway modulation and anticancer outcomes. It would strengthen the paper to explicitly state how the known antioxidant properties of these compounds are hypothesized or proven to contribute to these specific molecular modulations.
   • The paper needs to bridge the gap more consistently between a compound being an antioxidant andhow this antioxidant capacity translates into the observed effects on specific molecular targets like TGF-ß, Wnt, PI3K/Akt, etc., beyond general ROS scavenging.

 

 

 

 

 

 

2. Depth vs. Breadth in "Molecular Insights":
   • The review covers many molecular pathways (Figure 2 is very comprehensive). However, for some pathways, the discussion ofhow specific "antioxidant" phytochemicals intervene is somewhat general. Perhaps selecting a few key pathways and detailing the interplay with antioxidant phytochemicals more deeply would be more impactful than a broader, sometimes more superficial, overview for all.

 

• For instance, how does the redox-modulating capacity of certain phytochemicals directly influence the phosphorylation status or activity of kinases in the PI3K/Akt or MAPK pathways, or transcription factors like NF-kB, beyond general statements?

 

3. Novelty and Synthesis:
   • As a review, it's important to offer a novel synthesis or critical perspective. While comprehensive, the manuscript could benefit from a more explicit statement of its unique contribution. Is there a particular argument being advanced about the primacy of antioxidant effects or a novel framework for integrating these compounds into therapy based on their antioxidant and pathway-modulating effects? The section on immunogenic cell death (ICD) is a good example of a more novel angle.

Minor Issues:

1. Language and Clarity:
   • Abstract: "One of the most common and difficult cancers to treat that affects women globally is breast cancer." Could be rephrased for better flow, e.g., "Breast cancer is one of the most common and difficult-to-treat cancers affecting women globally."
   • Abstract: "We also go over how they affect..." is informal. Suggest "The review also discusses their effects on..."
   • Page 1, Abstract: "The article describes the difficulties..." could be "The review describes..." for consistency.
   • There are occasional awkward phrasings or minor grammatical errors that could be polished by careful proofreading. For example, page 6, line 235: "e.g. 3. superoxide dismutase..." and "e.g. 3. scavenge free radicals..."." — The “3" seems like a typo.
2. Figure/Table Specifics:
   • Tables 1 & 2 are very useful. Consider ensuring all IC50 values have consistent units or are clearly defined if mixed (e.g., µM vs µg/mL).
   • The legend for Figure 1 is quite long and incorporates references ([58, 59], [63], [69]). While informative, consider if some of this detail could be integrated into the main text to make the legend more concise, focusing primarily on describing the figure components.
3. Redundancy:
   • There's some overlap in the description of phytochemical actions across different sections (e.g., apoptosis and cell cycle arrest mentioned in the introduction, then again in Section 5, and summarized in Section 6). While some repetition is natural in a review, ensure it's purposeful for emphasis rather than just reiteration.
4. Specificity of Mechanisms:
   • In Tables 1 and 2, mechanisms like "induce apoptosis" are common. Where known and concise, adding a brief note on the key mediator (e.g., "via p53 activation" or "caspase-dependent") could enhance specificity, though the tables do already list some key proteins involved.
5. Justification of "Antioxidative Phytochemicals" in Section 5 Title:
   • The title of Section 5 is "Antioxidative Phytochemicals as Molecular Modulators..." The examples provided (resveratrol, hesperidin, and Rg5 via nanoparticles) are indeed often cited for antioxidant properties. However, the discussion in this section focuses more on their pathway modulation (YAP/TAZ, tumor microenvironment, drug delivery). As mentioned in Major Issue 1, strengthening the explicit link tohow their antioxidative nature contributes to these modulations would better justify the title.

Specific Comments by Page/Section:

• Page 1, Abstract, Lines 32-34:"By focusing on important signalling cascades like TGF-ẞ, Wnt...and estrogen receptor pathways, a variety of phytochemicals...demonstrate strong anticancer effects." Consider explicitly mentioning how their antioxidant nature contributes to these effects, if that is a primary argument.
• Page 6, Section 3:This section is well-focused on antioxidants. The challenge is to maintain this focus or clearly link back to it in subsequent sections discussing broader molecular effects.
• Page 12, Section 5:When discussing flavonoids, alkaloids, etc., and their alteration of PI3K/Akt, MAPK, and NF-kB, the text states they "prevent cell cycle progression and trigger apoptosis." It would be beneficial here to weave in how their antioxidative properties contribute to these specific outcomes on these pathways.
• Page 18, Section 6:This section summarizes "Antioxidant Functional Effects." Ensure this section truly emphasizes the "antioxidant" aspect of these functional effects rather than just the effects themselves (apoptosis, etc.), which could be mediated by non-antioxidant mechanisms as well.

Author Response

Reviewer Comments: 1

This review manuscript provides a comprehensive overview of the potential of plant-derived compounds (phytochemicals) in breast cancer treatment, with a specific focus on their molecular mechanisms and antioxidative properties. The authors cover a broad range of topics, including the historical context of phytotherapy, key oncogenic pathways in breast cancer, specific phytochemical classes and their actions, the role of antioxidants, and challenges in clinical translation. The inclusion of detailed figures illustrating molecular pathways (Fig. 2) and specific examples of phytochemical actions with experimental data (Figs. 3, 4, and 5), as well as informative tables summarizing compound activities (Tables 1 and 2), are definite strengths.

The manuscript is ambitious in its scope and generally well-structured. However, while "antioxidative" is in the title and the journal's focus, the direct and primary link between the antioxidant properties of many discussed compounds and their specific anticancer molecular mechanisms could be more explicitly and consistently elaborated throughout the text. In some sections, the antioxidant aspect feels secondary to the broader anticancer effects. The manuscript would benefit from a clearer delineation of when antioxidant activity is the primary mechanism versus a contributory one and how this specifically influences the discussed molecular pathways. Further polishing of the language and ensuring consistent focus on the "antioxidative therapeutic potentials" will enhance its impact.

Response:

We sincerely thank the reviewer for the thoughtful and constructive feedback on our manuscript. We are grateful for your recognition of the comprehensive nature of our review, the quality of the figures and tables, and the clarity of structure. Your comment regarding the need to more explicitly link the antioxidant properties of phytochemicals with their anticancer molecular mechanisms is well noted and appreciated.

In response to your valuable suggestion, we have made the following revisions:

1. We revised several sections of the manuscript to more clearly and consistently highlight the role of antioxidant activity as either a primary or contributory mechanism in the anticancer effects of phytochemicals. Specifically, we now explicitly indicate when oxidative stress modulation is central to pathway regulation (e.g., in the NF-κB or MAPK signaling cascades), and when it serves as a supportive mechanism alongside apoptosis induction or cell cycle arrest.

• Section 5, lines 341 – 364.
• Section 5, lines 386 – 413.

1. We added clarifying statements throughout the discussion and mechanism-related sections (notably in Sections 4 and 5) to delineate how the antioxidant properties influence the regulation of specific oncogenic and tumor-suppressive molecular pathways.

Section 4, lines 273 – 274.

Section 4, lines 298 – 306.

• Section 5, lines 386 – 413.

 

 

 

1. The title and abstract were slightly revised to reflect a sharper emphasis on the antioxidant-related therapeutic potential in order to align more closely with the journal's focus and the main message of the manuscript.

• Abstrct, lines 35 – 36.
• Abstrct, lines 45 – 49.

1. The language throughout the manuscript was further refined and polished to improve clarity and maintain a consistent emphasis on "antioxidative therapeutic potentials" where applicable.

We believe these improvements will enhance the clarity and scientific value of the manuscript, and we are grateful for your insightful recommendation to reinforce this central theme. Thank you once again for helping us improve the manuscript.

 

 

Major comments

Comments 1: Elaboration on the "Antioxidative" Link:

• While Section 3 directly addresses "Therapeutic Potentials of Antioxidants," the subsequent sections (e.g., Section 5 on "Antioxidative Phytochemicals as Molecular Modulators") discuss various phytochemicals and their effects on signalling pathways, but thespecific role of their antioxidant propertiesin modulating these pathways is not always clearly articulated. For example, when discussing resveratrol's effect on YAP/TAZ (Figure 3) or hesperidin's effects (Figure 4), the text describes pathway modulation and anticancer outcomes. It would strengthen the paper to explicitly state how the known antioxidant properties of these compounds are hypothesized or proven to contribute to these specific molecular modulations.
• The paper needs to bridge the gap more consistently between a compound being an antioxidant andhowthis antioxidant capacity translates into the observed effects on specific molecular targets like TGF-ß, Wnt, PI3K/Akt, etc., beyond general ROS scavenging.

Response 1:

Thank you for pointing this out. We agree with the reviewer’s insightful comments. To address this, we have revised the manuscript to more clearly articulate how the antioxidant properties of specific phytochemicals contribute to their modulation of molecular signaling pathways. Specifically, we have elaborated on the mechanistic links between antioxidant capacity and pathway modulation in the revised Section 5, particularly in the discussion of resveratrol. These changes can be found on Page 13, Paragraph 2.

We have added the following clarifications in the revised manuscript:

 

“Resveratrol, a polyphenol with potent antioxidant capacity, mitigates oxidative stress-induced activation of the YAP/TAZ pathway, likely through reduction of ROS-mediated upstream mechanical signals and redox-sensitive kinase activity.”

• Section 6, lines 415 – 418.

These revisions aim to explicitly link the antioxidative nature of these compounds to their observed effects on key signaling cascades such as YAP/TAZ, thus bridging the gap between their biochemical properties and molecular functions.

 

Comments 2: Depth vs. Breadth in "Molecular Insights":

• The review covers many molecular pathways (Figure 2 is very comprehensive). However, for some pathways, the discussion ofhowspecific "antioxidant" phytochemicals intervene is somewhat general. Perhaps selecting a few key pathways and detailing the interplay with antioxidant phytochemicals more deeply would be more impactful than a broader, sometimes more superficial, overview for all.
• For instance, how does the redox-modulating capacity of certain phytochemicals directly influence the phosphorylation status or activity of kinases in the PI3K/Akt or MAPK pathways, or transcription factors like NF-kB, beyond general statements?

 

Response 2:

We appreciate the reviewer’s valuable suggestion regarding the balance between depth and breadth in the discussion of molecular pathways. We agree that providing a more detailed analysis of selected pathways would enhance the impact and clarity of the review.

Therefore, we have revised Section 5 by focusing in greater depth on the PI3K/Akt, MAPK, pathways. We selected these as representative example and have elaborated on how the redox-modulating capacity of specific phytochemicals—such as curcumin,—influences kinase phosphorylation status and transcription factor activity through defined mechanisms.

The following content has been added to the manuscript:

• Section 5, lines 364 – 366.

“Curcumin has been shown to suppress the PI3K/Akt pathway by enhancing PTEN activity through oxidative stress reduction, thereby decreasing Akt phosphorylation,”

These targeted elaboration aim to provide a clearer mechanistic link between antioxidant phytochemicals and their modulation of key molecular pathways.

 

Comments 3: Novelty and Synthesis:

• As a review, it's important to offer a novel synthesis or critical perspective. While comprehensive, the manuscript could benefit from a more explicit statement of its unique contribution. Is there a particular argument being advanced about the primacy of antioxidant effects or a novel framework for integrating these compounds into therapy based on their antioxidant and pathway-modulating effects? The section on immunogenic cell death (ICD) is a good example of a more novel angle.

 

Response 3:

Thank you for pointing this out. We agree with the reviewer’s comment regarding the importance of articulating a clear and novel perspective in a review article. In response, we have revised the Introduction and Conclusion sections to explicitly state the unique contribution of this manuscript. Specifically, we now emphasize our central argument that the antioxidant properties of phytochemicals serve not merely as general cytoprotective mechanisms, but as strategic modulators of specific redox-sensitive signaling pathways, offering a targeted therapeutic avenue in cancer treatment.

Additionally, we have expanded the discussion in the Conclusion to propose a novel integrative framework that categorizes phytochemicals based on their dual role in redox regulation and pathway modulation, highlighting their potential for rational combination therapies.

The following content has been added in red:

“This review advances the perspective that antioxidant phytochemicals should not be viewed solely as passive ROS scavengers, but as active modulators of redox-sensitive signaling pathways (e.g., NF-κB, PI3K/Akt, and MAPK), where redox status directly impacts kinase activity, transcriptional regulation, and cell fate decisions in cancer.” (Page 3)

• Abstrct, lines 45 – 49.

“We propose a novel integrative framework in which antioxidant phytochemicals are classified according to their capacity to modulate distinct redox-sensitive molecular cascades. This synthesis highlights therapeutic windows where these compounds can synergize with conventional cancer therapies, particularly in contexts like immunogenic cell death (ICD) where redox balance is critical.”

• Section 2, lines 200 – 205.

Minor Issues:

Comments 1:Language and Clarity:

 

• Abstract: "One of the most common and difficult cancers to treat that affects women globally is breast cancer." Could be rephrased for better flow, e.g., "Breast cancer is one of the most common and difficult-to-treat cancers affecting women globally."
• Abstract: "We also go over how they affect..." is informal. Suggest "The review also discusses their effects on..."
• Page 1, Abstract: "The article describes the difficulties..." could be "The review describes..." for consistency.
• There are occasional awkward phrasings or minor grammatical errors that could be polished by careful proofreading. For example, page 7, line 278-279: "e.g. 3. superoxide dismutase..." and "e.g. 3. scavenge free radicals..."." — The “4" seems like a typo.

Response 1:

Thank you for pointing this out. We agree with the reviewer’s comments regarding language and clarity. Accordingly, we have revised several sentences in the Abstract and throughout the manuscript to improve flow, formal tone, and grammatical accuracy. These changes include rewording for clarity, correcting informal expressions, and fixing typographical errors.

Specifically, the following revisions were made:

Page 1, line 35-36.

“Breast cancer is one of the most common and difficult-to-treat cancers affecting women globally.”

Page 1, line 45-49.

“This review also discusses how they affect immune modulation, angiogenesis, cell cycle regulation, and apoptosis. Moreover, it also emphasis the challenges with these natural compounds' bioavailability, standardization, and clinical translation while highlighting preclinical and clinical research that supports their therapeutic potential.”

Page 31, line 278-279.

“They are endogenous antioxidants (e.g. superoxide dismutase, glutathione), as well as exogenous (e.g. vitamins C and E, flavonoids, and polyphenols), which scavenge free rad-icals and lessen oxidative stress.”

 

Comments 2:Figure/Table Specifics:

 

• Tables 1 & 2 are very useful. Consider ensuring all IC50 values have consistent units or are clearly defined if mixed (e.g., µM vs µg/mL).
• The legend for Figure 1 is quite long and incorporates references ([58, 59], [63], [69]). While informative, consider if some of this detail could be integrated into the main text to make the legend more concise, focusing primarily on describing the figure components.

 

Response 2:

Thank you for pointing this out. We agree with the reviewer’s helpful observations.

To address the first point, we have reviewed Tables 1 and 2 and ensured that all IC₅₀ values now include consistent units, either in µM or µg/mL, as appropriate. Where different units are used due to source variations, we have clearly indicated them in the table footnotes.

These updates can be found in Table and Table 2.

We believe these adjustments improve the manuscript’s clarity and presentation while preserving the informative value of the tables and figures.

 

 

Comments 3:Redundancy:

• There's some overlap in the description of phytochemical actions across different sections (e.g., apoptosis and cell cycle arrest mentioned in the introduction, then again in Section 5, and summarized in Section 6). While some repetition is natural in a review, ensure it's purposeful for emphasis rather than just reiteration.

 

Response 3:

Thank you for pointing this out. We agree with the reviewer’s comment regarding redundancy in the discussion of phytochemical actions across different sections.

To address this, we have revised the manuscript to minimize unnecessary repetition while maintaining important thematic emphasis. Specifically, we have streamlined the Introduction by removing redundant details on apoptosis and cell cycle arrest that are already discussed in depth in Section 5.  Additionally, we have revised Section 6 (Summary and Perspectives) to ensure it provides a concise synthesis without reiterating mechanistic details already presented earlier.

Revised content includes:

“These mechanistic details—such as induction of apoptosis and regulation of the cell cycle—are further elaborated in Section 5 in relation to specific signaling pathways.” (Page 2)

• Section 1, lines 97 – 99.

These edits aim to enhance the logical flow and cohesion of the review by reducing redundancy while preserving clarity and emphasis where appropriate.

 

 

Comments 4:Specificity of Mechanisms:

 

• In Tables 1 and 2, mechanisms like "induce apoptosis" are common. Where known and concise, adding a brief note on the key mediator (e.g., "via p53 activation" or "caspase-dependent") could enhance specificity, though the tables do already list some key proteins involved.

Response 4:

Thank you for pointing this out. We agree with the reviewer’s suggestion to enhance the specificity of the mechanisms described in Tables 1 and 2.

Accordingly, we have updated the tables to include brief but relevant mechanistic notes for entries where concise molecular mediators are known. For instance, instead of general terms like "induces apoptosis," we now indicate key mediators such as "via p53 activation," "caspase-3 dependent," or "through mitochondrial pathway" where supported by the cited literature.

• Section 4, lines 324 – 329.

 

 

Comments 5: Justification of "Antioxidative Phytochemicals" in Section 5 Title:

 

• The title of Section 5 is "Antioxidative Phytochemicals as Molecular Modulators..." The examples provided (resveratrol, hesperidin, and Rg5 via nanoparticles) are indeed often cited for antioxidant properties. However, the discussion in this section focuses more on their pathway modulation (YAP/TAZ, tumor microenvironment, drug delivery). As mentioned in Major Issue 1, strengthening the explicit link tohow their antioxidative nature contributesto these modulations would better justify the title.

Response 5:

Thank you for pointing this out. We agree with the reviewer’s observation that Section 5's title emphasizes the antioxidative nature of the phytochemicals, while the discussion focused more heavily on their pathway-modulating effects.

To address this, we have revised the content in Section 5, to explicitly clarify how the antioxidant properties of compounds like resveratrol, hesperidin, and ginsenoside Rg5 contribute to their modulation of specific molecular pathways. This includes referencing their role in reducing intracellular ROS levels, which impacts signaling cascades such as PI3K/Akt, YAP/TAZ, and NF-κB, thereby influencing cancer cell behavior.

 “Resveratrol’s antioxidative capacity helps suppress oxidative stress-mediated activation of YAP/TAZ signaling, thereby reducing oncogenic transcriptional activity (Page 13, Paragraph 2, Line 425).”

“Hesperidin reduces ROS accumulation, which plays a key upstream role in PI3K/Akt pathway attenuation and apoptosis induction (Page 16, Paragraph 3, Line 523-525).”

“The antioxidative function of ginsenoside Rg5 contributes to remodeling the tumor microenvironment by decreasing oxidative stress, which modulates immune suppression and angiogenic signaling (Page 19, lines 580 - 582).”

These clarifications strengthen the justification for the current section title:

“Antioxidative Phytochemicals as Molecular Modulators of Breast Cancer Pathways.”

 

Specific Comments by Page/Section:

Comments 1:

 

Page 1, Abstract, Lines 32-34:"By focusing on important signalling cascades like TGF-ẞ, Wnt...and estrogen receptor pathways, a variety of phytochemicals...demonstrate strong anticancer effects." Consider explicitly mentioning how their antioxidant nature contributes to these effects, if that is a primary argument.

 

Response 1:

Thank you for highlighting this point. We agree that clarifying the connection between the antioxidant properties of phytochemicals and their modulation of signaling pathways would strengthen the abstract and reinforce the central theme of the manuscript.

“By focusing on important signalling cascades like TGF-β, Wnt, PI3K/Akt, and estrogen receptor pathways, a variety of phytochemicals with well-established antioxidant properties demonstrate strong anticancer effects, partly by mitigating oxidative stress that drives these oncogenic pathways.”

 

 

 

Comments 2:

Page 7, Section 4: This section is well-focused on antioxidants. The challenge is to maintain this focus or clearly link back to it in subsequent sections discussing broader molecular effects.

Response 2:

Thank you for this valuable observation. We agree that maintaining a clear and consistent connection to the antioxidant theme throughout the manuscript enhances coherence and strengthens the central narrative.

 

To address this, we have revised the transitions and introductory sentences of Sections 5 and 6 to explicitly link their broader molecular discussions back to the antioxidant functions of the phytochemicals.

 

Revised content in red includes:

 

Page 7, Section 4, Paragraph 1, Line 271-274:

“Building upon the antioxidant mechanisms outlined in Section , this section explores how phytochemicals influence apoptosis, cell cycle regulation, and other hallmarks of cancer.”

 

Page 11, Section 5, Paragraph 1, Line 339-341:

“These phytochemicals often exert their modulatory effects on signaling pathways in part through their antioxidant action, which reduces oxidative stress and alters redox-sensitive molecular cascades.”

 

These revisions help maintain thematic continuity and reinforce the central role of antioxidant activity in the anticancer mechanisms discussed throughout the review.

 

 

Comments 3:

Page 12, Section 5:When discussing flavonoids, alkaloids, etc., and their alteration of PI3K/Akt, MAPK, and NF-kB, the text states they "prevent cell cycle progression and trigger apoptosis." It would be beneficial here to weave in how their antioxidative properties contribute to these specific outcomes on these pathways.

 

Response 3:

Thank you for this insightful comment. We agree that explicitly linking the antioxidative properties of flavonoids, alkaloids, and other phytochemicals to their modulation of PI3K/Akt, MAPK, and NF-κB pathways will enhance the mechanistic clarity.

Therefore, we have revised the relevant text in Section 5 (Page 13, Paragraph 1, Lines 424–428) to incorporate how the antioxidant activity reduces intracellular reactive oxygen species (ROS), which in turn influences the phosphorylation status and activity of key kinases and transcription factors involved in cell cycle arrest and apoptosis induction.

“The antioxidative properties of flavonoids and alkaloids contribute to their ability to prevent cell cycle progression and trigger apoptosis by lowering ROS levels that otherwise activate PI3K/Akt, MAPK, and NF-κB signaling. This reduction in oxidative stress modulates the phosphorylation and activity of these pathways, ultimately promoting cancer cell death.”

This addition clarifies the causal role of antioxidant activity in mediating these molecular effects.

 

 

 

 

Comments 4:

 

Page 18, Section 7: This section summarizes "Antioxidant Functional Effects." Ensure this section truly emphasizes the "antioxidant" aspect of these functional effects rather than just the effects themselves (apoptosis, etc.), which could be mediated by non-antioxidant mechanisms as well.

Response 4:

Thank you for your insightful comment. We agree that Section 6 should emphasize the antioxidant basis of the functional effects summarized, to maintain thematic consistency and accurately reflect the focus of the review.

To address this, we have revised Section 7 (Page 18, Paragraph 2) to explicitly highlight how the antioxidant properties of phytochemicals underpin key functional outcomes such as apoptosis, cell cycle arrest, and immune modulation. The revised text clarifies that while some effects might involve additional mechanisms, antioxidant activity plays a central and foundational role.

Revised text:

“The functional effects of phytochemicals—including induction of apoptosis, cell cycle regulation, and modulation of immune responses—are often driven primarily by the antioxidative capacity of phytochemicals, which mitigates oxidative stress and thereby influences redox-sensitive signaling pathways.”

Page 19, Section 7, Line 601-604:

This clarification strengthens the section’s focus on the antioxidant dimension of these therapeutic effects.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The authors explain how phytotherapy could be useful for treating breast cancer by detailing the mechanisms of action phytochemicals have toward its inhibition, such as controlling the immune system, reducing cellular multiplication, and inducing apoptosis. From the numerous groups of phytochemicals, the authors focus on flavonoids and polyphenols and elaborate on their antioxidant and antitumor effects while explaining some of the experimental differences of some phytochemicals, such as hesperidin and resveratrol. Due to the difficulties concerning application of phytotherapy in clinical settings, the authors analyze some of the clinical gaps such as the clinical study research, the bioavailability, and the standards of practice.

 

The mechanisms of action for these natural phytochemicals including akaloids, terpenoids, polyphenol and flavonoid on major molecular pathways/processes regulated during breast carcinogenesis and treatment such as: (1) cell proliferation and survival pathways, (2) metastasis, invasion, and transformation, (3) apoptosis and cell cycle, (4) angiogenesis, (5) immune modulation, (6) oxidative stress, (7) inflammation, and (8) drug resistance, have been summarized by the authors. The authors emphasized that breast cancer is influenced by phytochemicals of natural origin and their mechanisms of action are multifaceted and concordant, and they span from cell signalling and gene expression, to cell cycle progression, apoptosis and immunoregulation. Their multitargeting feature provides a promising application for a complete treatment of breast cancer.

 

Extensive in vitro studies have demonstrated that phytochemicals possess important pharmacological properties, including anti-inflammatory, anti-cancer, and antioxidant effects. These studies provide mechanistic insights into how phytochemicals exert their therapeutic effects at the cellular and molecular levels.

Specifically, in vitro studies have shown that certain antioxidants (e.g., curcumin, resveratrol, quercetin, and EGCG) can modulate key signaling pathways, such as PI3K/Akt, MAPK, and NF-κB, which play key roles in inflammation, metastasis, and cell survival in breast cancer.

The authors reviewed preclinical and clinical studies to investigate the use of phytotherapy as an adjunctive treatment for breast cancer in a variety of ways:

1. Elucidating Mechanisms of Action:

A number of in vitro investigations have demonstrated that phytochemicals have the ability to stop the cell cycle and trigger apoptosis, or programmed cell death, two important processes for preventing the growth of tumors. For instance, it has been demonstrated that quercetin, genistein, and apigenin cause apoptosis and halt the progression of the cell cycle in breast cancer cells. According to the authors, resveratrol (REV) has been demonstrated in vitro to inhibit YAP/TAZ signaling in breast cancer cells, which lowers the expression of YAP target genes and prevents cell invasion and RhoA activation. According to the MTT assay, REV did not appear to be harmful to cells at specific concentrations.

2. Evaluating In Vivo Efficacy and Safety:

Preclinical in vivo research, often using animal models, further supports the therapeutic potential of phytotherapy. These studies provide key translational insights into the systemic bioavailability, pharmacokinetics, and organ-specific responses of phytochemicals in vivo, which are critical for clinical development. The authors mentioned two phytochemicals, including hesperidin and ginsenoside Rg5. The results showed that both significantly reduced the breast tumor xenograft in the mouse model. Especially the Rg5, which was conjugated with nanoparticles to improve the toxic effects of Rg5 on breast tumors in an in vivo system.

3. Transitioning to Human Clinical Application:

As mentioned in this work, in recent years, clinical research has started to find out how phytochemicals, when used on their own or together with other types of therapy, can be helpful. For example, test was done to see whether curcumin and genistein could make cancer medicines work better and make their effects less harmful. Moreover, we found that resveratrol may recombine with tamoxifen on this way of curbing misfit in breast cancer. This clinical work, in spite of being needed to make sure that such work can be done on humans, is very important to lead us to find more ways for translation of phytotherapy into the field of evidence-based practice of treatment of cancer.

The authors argued that preclinical (in vitro and in vivo animal) studies provide solid evidence of molecular mechanisms and biological effects for the potential benefits of phytotherapy in breast cancer treatment, including inhibition of tumor growth, induction of cell death, modulation of immunity and angiogenesis. Preliminary and ongoing clinical trials are beginning to translate these findings into safety and efficacy data for human use, providing support for the use of phytotherapy as an adjunct treatment approach. Although there are still challenges in bioavailability, standardization and the need for more rigorous clinical validation, the existing research results are an important basis to support its application.

 

In addition, the authors also mentioned that although phytotherapy shows promising potential in breast cancer treatment, it still faces some important challenges in clinical application. These challenges mainly focus on the following aspects:

1. Bioavailability

This is one of the main obstacles to the clinical application of phytochemicals

. Problems with many phytochemicals include poor water solubility, limited systemic absorption, rapid metabolism, limited systemic circulation, and nonspecific biodistribution.

2. Standardization

The authors point out that variations in plant origin, growing circumstances, and extraction methods result in inherent heterogeneity in phytochemical composition. It is challenging to guarantee consistent treatment results because of this diversity. Therefore, to guarantee reproducibility and dependability in the clinical context, standardized processes must be implemented in the areas of dose optimization, quality assurance, and raw material procurement.

3. Clinical Translation and Validation

More thorough preclinical and clinical evaluation is needed to elucidate the safety, therapeutic index, and possible interactions of phytotherapy with conventional chemotherapeutic agents. Some clinical trials have reported conflicting or inconclusive results. Additional large-scale randomized controlled trials are needed to verify its therapeutic benefit and address variability in patient response. For example, while heparin flavonoids have shown promising anti-tumor potential in mouse models, the data indicate that additional translational studies are needed to evaluate their clinical efficacy

4. Potential Interference: Herbal therapy may potentially disrupt the way that traditional therapy works. For instance, antioxidants may disrupt the harmful pathways mediated by reactive oxygen species (ROS) in conventional treatments. Last but not least, the authors contended that despite the fact that phytochemicals have a number of potential therapeutic advantages for breast cancer, their use is restricted by low bioavailability, challenges with standardizing formulations, and the requirement for more thorough clinical validation to ascertain safe dosage, effectiveness, and interactions with traditional therapies. Incorporating phytotherapy into all-encompassing cancer treatment plans requires overcoming these obstacles.

     There are still many related issues that I haven’t mentioned above. From my point of view, this is an extensive review article regarding the phytochemicals and breast cancer treatment. There is a large quantity of valuable information classified and summarized by the authors, who are also experts in the related fields. The writing and logic are adequate. However, some similar reviewed articles were published before, which might weaken the significance and sound of this article. To strengthen the scientific value and distinctiveness, the authors might add a discussion about which pathway was most affected by phytochemicals, or ranking the pathways affected by phytochemicals, or the effects and underlying mechanisms of combined phytochemicals in treating breast cancer. Besides, the resolution of the figures is low, which makes them look blurred.

Author Response

Reviewer Comments: 2

 

Comments 1: Authors should add the specific plant names targeted in this review for better presentation, and the same can be elaborated in detail in the introduction section.

Response 1:

Thank you for pointing this out. We agree that specifying the plant names targeted in this review will improve clarity and presentation. Therefore, we have added a detailed mention of the specific plants discussed in this review in the Introduction section.

“This review focuses on phytochemicals derived from plants such as Arenga porphyrocarpa, Vitis vinifera (grapevine), Citrus sinensis (orange), and Panax ginseng, highlighting their antioxidant properties and molecular mechanisms relevant to anticancer effects.”

Page No. 3, Lines 134-137.

This addition provides readers with clearer context on the botanical sources central to the manuscript.

Comments 2:

The authors explain how phytotherapy could be useful for treating breast cancer by detailing the mechanisms of action phytochemicals have toward its inhibition, such as controlling the immune system, reducing cellular multiplication, and inducing apoptosis. From the numerous groups of phytochemicals, the authors focus on flavonoids and polyphenols and elaborate on their antioxidant and antitumor effects while explaining some of the experimental differences of some phytochemicals, such as hesperidin and resveratrol. Due to the difficulties concerning application of phytotherapy in clinical settings, the authors analyze some of the clinical gaps such as the clinical study research, the bioavailability, and the standards of practice.

The mechanisms of action for these natural phytochemicals including akaloids, terpenoids, polyphenol and flavonoid on major molecular pathways/processes regulated during breast carcinogenesis and treatment such as: (1) cell proliferation and survival pathways, (2) metastasis, invasion, and transformation, (3) apoptosis and cell cycle, (4) angiogenesis, (5) immune modulation, (6) oxidative stress, (7) inflammation, and (8) drug resistance, have been summarized by the authors. The authors emphasized that breast cancer is influenced by phytochemicals of natural origin and their mechanisms of action are multifaceted and concordant, and they span from cell signalling and gene expression, to cell cycle progression, apoptosis and immunoregulation. Their multitargeting feature provides a promising application for a complete treatment of breast cancer.

Extensive in vitro studies have demonstrated that phytochemicals possess important pharmacological properties, including anti-inflammatory, anti-cancer, and antioxidant effects. These studies provide mechanistic insights into how phytochemicals exert their therapeutic effects at the cellular and molecular levels.

Specifically, in vitro studies have shown that certain antioxidants (e.g., curcumin, resveratrol, quercetin, and EGCG) can modulate key signaling pathways, such as PI3K/Akt, MAPK, and NF-κB, which play key roles in inflammation, metastasis, and cell survival in breast cancer.

The authors reviewed preclinical and clinical studies to investigate the use of phytotherapy as an adjunctive treatment for breast cancer in a variety of ways:

1. Elucidating Mechanisms of Action:

A number of in vitro investigations have demonstrated that phytochemicals have the ability to stop the cell cycle and trigger apoptosis, or programmed cell death, two important processes for preventing the growth of tumors. For instance, it has been demonstrated that quercetin, genistein, and apigenin cause apoptosis and halt the progression of the cell cycle in breast cancer cells. According to the authors, resveratrol (REV) has been demonstrated in vitro to inhibit YAP/TAZ signaling in breast cancer cells, which lowers the expression of YAP target genes and prevents cell invasion and RhoA activation. According to the MTT assay, REV did not appear to be harmful to cells at specific concentrations.

2. Evaluating In Vivo Efficacy and Safety:

Preclinical in vivo research, often using animal models, further supports the therapeutic potential of phytotherapy. These studies provide key translational insights into the systemic bioavailability, pharmacokinetics, and organ-specific responses of phytochemicals in vivo, which are critical for clinical development. The authors mentioned two phytochemicals, including hesperidin and ginsenoside Rg5. The results showed that both significantly reduced the breast tumor xenograft in the mouse model. Especially the Rg5, which was conjugated with nanoparticles to improve the toxic effects of Rg5 on breast tumors in an in vivo system.

3. Transitioning to Human Clinical Application:

As mentioned in this work, in recent years, clinical research has started to find out how phytochemicals, when used on their own or together with other types of therapy, can be helpful. For example, test was done to see whether curcumin and genistein could make cancer medicines work better and make their effects less harmful. Moreover, we found that resveratrol may recombine with tamoxifen on this way of curbing misfit in breast cancer. This clinical work, in spite of being needed to make sure that such work can be done on humans, is very important to lead us to find more ways for translation of phytotherapy into the field of evidence-based practice of treatment of cancer.

The authors argued that preclinical (in vitro and in vivo animal) studies provide solid evidence of molecular mechanisms and biological effects for the potential benefits of phytotherapy in breast cancer treatment, including inhibition of tumor growth, induction of cell death, modulation of immunity and angiogenesis. Preliminary and ongoing clinical trials are beginning to translate these findings into safety and efficacy data for human use, providing support for the use of phytotherapy as an adjunct treatment approach. Although there are still challenges in bioavailability, standardization and the need for more rigorous clinical validation, the existing research results are an important basis to support its application.

 

In addition, the authors also mentioned that although phytotherapy shows promising potential in breast cancer treatment, it still faces some important challenges in clinical application. These challenges mainly focus on the following aspects:

1. Bioavailability

This is one of the main obstacles to the clinical application of phytochemicals

. Problems with many phytochemicals include poor water solubility, limited systemic absorption, rapid metabolism, limited systemic circulation, and nonspecific biodistribution.

2. Standardization

The authors point out that variations in plant origin, growing circumstances, and extraction methods result in inherent heterogeneity in phytochemical composition. It is challenging to guarantee consistent treatment results because of this diversity. Therefore, to guarantee reproducibility and dependability in the clinical context, standardized processes must be implemented in the areas of dose optimization, quality assurance, and raw material procurement.

3. Clinical Translation and Validation

More thorough preclinical and clinical evaluation is needed to elucidate the safety, therapeutic index, and possible interactions of phytotherapy with conventional chemotherapeutic agents. Some clinical trials have reported conflicting or inconclusive results. Additional large-scale randomized controlled trials are needed to verify its therapeutic benefit and address variability in patient response. For example, while heparin flavonoids have shown promising anti-tumor potential in mouse models, the data indicate that additional translational studies are needed to evaluate their clinical efficacy

4. Potential Interference: Herbal therapy may potentially disrupt the way that traditional therapy works. For instance, antioxidants may disrupt the harmful pathways mediated by reactive oxygen species (ROS) in conventional treatments. Last but not least, the authors contended that despite the fact that phytochemicals have a number of potential therapeutic advantages for breast cancer, their use is restricted by low bioavailability, challenges with standardizing formulations, and the requirement for more thorough clinical validation to ascertain safe dosage, effectiveness, and interactions with traditional therapies. Incorporating phytotherapy into all-encompassing cancer treatment plans requires overcoming these obstacles.

 There are still many related issues that I haven’t mentioned above. From my point of view, this is an extensive review article regarding the phytochemicals and breast cancer treatment. There is a large quantity of valuable information classified and summarized by the authors, who are also experts in the related fields.

The writing and logic are adequate. However, some similar reviewed articles were published before, which might weaken the significance and sound of this article. To strengthen the scientific value and distinctiveness, the authors might add a discussion about which pathway was most affected by phytochemicals, or ranking the pathways affected by phytochemicals, or the effects and underlying mechanisms of combined phytochemicals in treating breast cancer. Besides, the resolution of the figures is low, which makes them look blurred.

Response 3:

We sincerely thank the reviewer for the thoughtful and constructive feedback on our manuscript. We are grateful for your recognition of the comprehensive nature of our review, the quality of the figures and tables, and the clarity of structure. Your comment regarding the need to more explicitly link the antioxidant properties of phytochemicals with their anticancer molecular mechanisms is well noted and appreciated.

In response, we have made the following revisions:

We Added a dedicated subsection in the Discussion (Page 7, Paragraph 2, lines 305-313 ) where we analyze and rank the major molecular pathways modulated by phytochemicals based on the current literature. This section highlights pathways such as PI3K/Akt, MAPK, NF-κB, apoptosis, immune modulation, and oxidative stress, emphasizing which are most consistently and strongly influenced by different phytochemical classes. This addition provides readers with clearer insight into the relative importance and multitargeting nature of these pathways in breast cancer therapy.

In response to the suggestion to rank pathways or discuss synergistic effects, we have incorporated a comparative summary in the discussion section, outlining the relative impact of phytochemicals on key pathways based on available preclinical and clinical evidence. Additionally, we have added content discussing the effects and potential synergistic mechanisms of phytochemical combinations, such as curcumin and genistein or resveratrol and tamoxifen, in breast cancer models.

Regarding the resolution of the figures, we thank the reviewer for pointing this out. All figures have now been replaced with high-resolution versions to ensure better clarity and readability in both print and digital formats.

We believe these modifications address your suggestions comprehensively and significantly strengthen the scientific depth and clarity of our review. We are grateful for your constructive feedback that helped improve our manuscript.

 

 

 

 

 

 

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

ajor Comments:

1. Clarity of  Introduction Regarding Specific Plants:

   • In the Introduction (Section 1, lines 134-136), the authors state: "This review focuses on phytochemicals derived from plants such as Arenga porphyrocarpa, Vitis vinifera (grapevine), Citrus sinensis (orange), and Panax ginseng, highlighting their antioxidant properties and molecular mechanisms relevant to anticancer effects." While these are good examples, the review (evident from Tables 1 & 2 and subsequent discussions) covers a much broader range of plants and phytochemicals. This specific mention in the introduction might unintentionally narrow the perceived scope for the reader.

   •  Please rephrase this sentence to clarify that these plants are examples of those discussed, rather than the sole or primary focus, to reflect the comprehensive nature of the review better. For instance: "...plants including, but not limited to, Arenga porphyrocarpa..." or "...plants, with examples such as Arenga porphyrocarpa..."

2. Figure 1 (Mechanisms of Action):

   • Figure 1 is referred to multiple times (e.g., page 4, line 171 for Figure 1(A); line 181 for Figure 1(B); line 188 for Figure 1(C); line 191 for Figure 1(D); page 5, line 200 for Figure 1(E)). However, the main schematic for Figure 1(A) ("Studies of the mechanisms of action of phytochemicals in breast cancer cells...") seems to be a general diagram showing plant extract to in vitro/in vivo studies leading to various regulatory outcomes. If there's a more detailed introductory figure planned that wasn't included in the OCR or is crucial for understanding the initial framework of phytochemical action, its absence could be confusing. The current Figure 1 caption on page 5 (lines 210-218) describes sub-panels (A) through (E) quite well. Assuming the figure on page 5 is the intended Figure 1, this is less of a significant issue, but please make sure all parts are presented and of high resolution.

3.  Methodology :

   • The methodology section describes a search strategy, keywords, databases, and inclusion/exclusion criteria, which is commendable. However, the term "review" can be broad. It might be beneficial to explicitly state that this is a comprehensive narrative review or literature review rather than a systematic review (which typically involves stricter PRISMA guidelines, risk of bias assessment, etc., not evident here). This is not a criticism of the approach but a suggestion for precision.

   •  Consider adding a sentence at the beginning of Section 3 to define the type of review (e.g., "This comprehensive narrative review aims to...").

Minor Comments:

1. Abstract (Line 47): "Moreover, it also emphasizes..." should be "Moreover, it also emphasizes..."

2. Figure 1(E) Caption Detail (Page 5, Line 203-204): "...decreasing immunosuppressive populations like myeloid-d [69]." It's likely "myeloid-d" refers to "myeloid-derived suppressor cells (MDSCs)" or a similar term. Please clarify or spell out for better understanding.

3. Figure Quality: Ensure all figures (especially complex diagrams like Figure 2 and the multi-panel Figures 3, 4, 5) are submitted at high resolution for optimal clarity in the final publication.

4. Table 2 Formatting (Page 10 onwards): The highlighted yellow text within Table 2 (e.g., under Epigallocatechin gallate, G28, Resveratrol) makes the table difficult to read and appears to be an artifact or author emphasis not suitable for final publication. Please ensure this highlighting is removed.

5. Consistency in Gene/Protein Nomenclature: Please ensure consistency in using italics for gene names and standard font for protein names throughout the manuscript, as per journal guidelines.

6. Author Contributions (Page 21, Lines 696-698):

   • "writing-original draft preparation, R.K.;" – The corresponding author for this initial is Raj Kumar Mongre (R.K.M.). Please verify if R.K.M. or another author was responsible for the original draft preparation and use the correct initials.

   • The contributions seem well-distributed otherwise.

7. Page 7, Section "4. Therapeutic Potentials of Antioxidants..." (Line 275): "Building upon the antioxidant mechanisms outlined in Section," – it might be clearer to specify which section (e.g., "outlined in Section X," or "outlined previously/above"). If it refers to general knowledge, it's acceptable.

8. Page 8, Line 324: "clinical validation is necessary."

9. Page 11, Line 342: "These phytochemicals often exert..."

10. Page 19, Line 587-589 (Figure 5B context): "The antioxidative function of ginsenoside Rg5 contributes to remodeling the tumor microenvironment by decreasing oxidative stress, which modulates immune suppression and angiogenic signaling." This is a clear and important statement.

11. References: The manuscript is well-referenced with a mix of recent and foundational papers. Ensure all citations are accurate and complete.

Strengths of the Paper:

• Comprehensive Coverage: The review thoroughly covers a wide array of phytochemicals, their sources, molecular targets, and mechanisms in the context of breast cancer.

• Mechanistic Insights: It delves into critical signaling pathways (e.g., PI3K/Akt, MAPK, Wnt, TGF-β, Notch, VEGF, HER2) and cellular processes (apoptosis, cell cycle, angiogenesis, immune modulation).

• Illustrative Material: Including detailed figures and tables effectively summarizes complex information and enhances understanding. Figures 3, 4, and 5, detailing specific compound studies, are particularly informative.

• Relevance and Timeliness: The topic of natural compounds in cancer therapy is highly relevant and of growing interest.

• Discussion of Challenges: The authors appropriately address significant hurdles like bioavailability, standardization, and the need for robust clinical translation.

• Clear Structure: The manuscript is logically organized, making it easy for readers to follow the progression of ideas.

 

Author Response

Reviewer Comments: 1

Comments 1: In the Introduction (Section 1, lines 134-136), the authors state: "This review focuses on phytochemicals derived from plants such as Arenga porphyrocarpa, Vitis vinifera (grapevine), Citrus sinensis (orange), and Panax ginseng, highlighting their antioxidant properties and molecular mechanisms relevant to anticancer effects." While these are good examples, the review (evident from Tables 1 & 2 and subsequent discussions) covers a much broader range of plants and phytochemicals. This specific mention in the introduction might unintentionally narrow the perceived scope for the reader.

Please rephrase this sentence to clarify that these plants are examples of those discussed, rather than the sole or primary focus, to reflect the comprehensive nature of the review better. For instance: "...plants including, but not limited to, Arenga porphyrocarpa..." or "...plants, with examples such as Arenga porphyrocarpa..."

Response 1: Thank you for your thoughtful observation. We appreciate your suggestion and agree that the original phrasing may have inadvertently implied a limited scope. To better reflect the comprehensive nature of our review, we have revised the sentence in the Introduction to read: “...plants, with examples such as Arenga porphyrocarpa, Vitis vinifera, Citrus sinensis, and Panax ginseng...” This adjustment ensures clarity and emphasizes that these are representative examples rather than an exhaustive or exclusive list, aligning with the broader content presented.

 

(Section 1, lines 134-137)

 

This review explores the antioxidant properties and anticancer mechanisms of phytochemicals derived from a diverse array of plants, with representative examples such as Arenga porphyrocarpa, Vitis vinifera (grapevine), Citrus sinensis (orange), and Panax ginseng.

 

Comments 2: Figure 1 (Mechanisms of Action):

Figure 1 is referred to multiple times (e.g., page 4, line 171 for Figure 1(A); line 181 for Figure 1(B); line 188 for Figure 1(C); line 191 for Figure 1(D); page 5, line 200 for Figure 1(E)). However, the main schematic for Figure 1(A) ("Studies of the mechanisms of action of phytochemicals in breast cancer cells...") seems to be a general diagram showing plant extract to in vitro/in vivo studies leading to various regulatory outcomes. If there's a more detailed introductory figure planned that wasn't included in the OCR or is crucial for understanding the initial framework of phytochemical action, its absence could be confusing. The current Figure 1 caption on page 5 (lines 210-218) describes sub-panels (A) through (E) quite well. Assuming the figure on page 5 is the intended Figure 1, this is less of a significant issue, but please make sure all parts are presented and of high resolution.

Response 2:

Thank you for your thoughtful observation. We appreciate your attention to detail regarding Figure 1. We confirm that the figure on page 5 is indeed the intended Figure 1, with sub-panels (A) through (E) clearly representing the framework of phytochemical action. We agree that the current caption provides clarity; however, we will ensure all figure parts are consistently referenced and of high resolution. Your comment has helped enhance the coherence and presentation of our manuscript—thank you again for your valuable feedback.

 

Comments 3: Methodology :

The methodology section describes a search strategy, keywords, databases, and inclusion/exclusion criteria, which is commendable. However, the term "review" can be broad. It might be beneficial to explicitly state that this is a comprehensive narrative review or literature review rather than a systematic review (which typically involves stricter PRISMA guidelines, risk of bias assessment, etc., not evident here). This is not a criticism of the approach but a suggestion for precision.

Consider adding a sentence at the beginning of Section 3 to define the type of review (e.g., "This comprehensive narrative review aims to...").

Response 3: We sincerely thank the reviewer for their thoughtful and constructive feedback regarding the clarification of the review type. We fully agree that distinguishing between a systematic review and a narrative or literature review enhances the methodological transparency and interpretability of the manuscript.

In response, we have explicitly clarified the nature of this work as a comprehensive narrative review at the beginning of Section 3. The revised sentence now reads:

"This comprehensive narrative review aims to compile and critically examine the molecular targets and mechanisms through which anti-inflammatory and antioxidant phytochemicals may prevent breast cancer."

This modification aligns with the scope and methodology employed, which, while systematic in literature search and selection, does not fully adhere to PRISMA guidelines or include a formal risk of bias assessment—hallmarks of a systematic review. We believe this clarification improves the precision of our methodology without altering the integrity of our approach.

Once again, we appreciate this valuable suggestion, which has helped to enhance the clarity and academic rigor of our manuscript.

 

Minor Comments:

1. Abstract (Line 47):"Moreover, it also emphasizes..." should be "Moreover, it also emphasizes..."
2. Figure 1(E) Caption Detail (Page 5, Line 203-204):"...decreasing immunosuppressive populations like myeloid-d [69]." It's likely "myeloid-d" refers to "myeloid-derived suppressor cells (MDSCs)" or a similar term. Please clarify or spell out for better understanding.
3. Figure Quality:Ensure all figures (especially complex diagrams like Figure 2 and the multi-panel Figures 3, 4, 5) are submitted at high resolution for optimal clarity in the final publication.
4. Table 2 Formatting (Page 10 onwards):The highlighted yellow text within Table 2 (e.g., under Epigallocatechin gallate, G28, Resveratrol) makes the table difficult to read and appears to be an artifact or author emphasis not suitable for final publication. Please ensure this highlighting is removed.
5. Consistency in Gene/Protein Nomenclature:Please ensure consistency in using italics for gene names and standard font for protein names throughout the manuscript, as per journal guidelines.
6. Author Contributions (Page 21, Lines 696-698):
   • "writing-original draft preparation, R.K.;" – The corresponding author for this initial is Raj Kumar Mongre (R.K.M.). Please verify if R.K.M. or another author was responsible for the original draft preparation and use the correct initials.
   • The contributions seem well-distributed otherwise.
7. Page 7, Section "4. Therapeutic Potentials of Antioxidants..." (Line 275):"Building upon the antioxidant mechanisms outlined in Section," – it might be clearer to specify which section (e.g., "outlined in Section X," or "outlined previously/above"). If it refers to general knowledge, it's acceptable.
8. Page 8, Line 324:"clinical validation is necessary."
9. Page 11, Line 342:"These phytochemicals often exert..."
10. Page 19, Line 587-589 (Figure 5B context):"The antioxidative function of ginsenoside Rg5 contributes to remodeling the tumor microenvironment by decreasing oxidative stress, which modulates immune suppression and angiogenic signaling." This is a clear and important statement.
11. References:The manuscript is well-referenced with a mix of recent and foundational papers. Ensure all citations are accurate and complete.

Response:

We sincerely thank the reviewer for the thorough and constructive feedback. We have carefully addressed each point raised to enhance the clarity, consistency, and overall quality of the manuscript. Please find our detailed responses below:

1. Abstract (Line 47)
   Comment: "Moreover, it also emphasizes..." should be "Moreover, it also emphasizes..."
   Response: We acknowledge this observation, though it appears the text remains unchanged. We have double-checked the sentence structure for clarity and grammar and confirm that it now reads appropriately in the revised manuscript.
2. Figure 1(E) Caption Detail (Page 5, Lines 203–204)
   Comment: Clarify “myeloid-d” as it likely refers to “myeloid-derived suppressor cells (MDSCs).”
   Response: Thank you for pointing this out. We have now corrected the term by spelling out "myeloid-derived suppressor cells (MDSCs)" in the figure caption for clarity and to avoid ambiguity.
3. Figure Quality
   Comment: Ensure high-resolution submission of all complex figures (Figures 2, 3, 4, 5).
   Response: We fully agree with the importance of high-resolution figures for clarity. All multi-panel and schematic figures have been replaced with high-resolution versions in the revised manuscript to ensure optimal legibility and publication quality.
4. Table 2 Formatting (Page 10 onwards)
   Comment: Remove highlighted yellow text, as it may be an artifact or unintended emphasis.
   Response: We appreciate this observation. The yellow highlighting was unintentional and has now been completely removed to maintain a professional and publication-ready format for Table 2.
5. Consistency in Gene/Protein Nomenclature
   Comment: Ensure italics for gene names and standard font for protein names.
   Response: Thank you for this important reminder. We have carefully reviewed the manuscript and revised gene/protein nomenclature throughout to align with the journal's formatting guidelines.
6. Author Contributions (Page 21, Lines 696–698)
   Comment: Clarify if "R.K." refers to "Raj Kumar Mongre (R.K.M.)" in original draft preparation.
   Response: We appreciate the reviewer’s attention to detail. This has been corrected in the revised version; the initials have been updated from "R.K." to "R.K.M." to accurately reflect Dr. Raj Kumar Mongre’s contribution to the original draft preparation.
7. Section Reference Clarity (Page 7, Line 275)
   Comment: Specify which section is being referred to or revise for clarity.
   Response: Thank you for the suggestion. We have revised the sentence to read: “Building upon the antioxidant mechanisms outlined in Section 3,” for improved clarity and ease of reference.

8–9. Clarity in Sentences (Page 8, Line 324; Page 11, Line 342)
Comment: Ensure clarity and impact in these specific statements.
Response: We appreciate the reviewer highlighting these lines. Both statements have been retained as they effectively convey the intended meaning, but we have slightly refined surrounding text for improved flow where necessary.

10. Page 19, Lines 587–589 (Figure 5B Context)
    Comment: This is a clear and important statement.
    Response: We are grateful for the positive feedback. We have retained this statement as is, recognizing its relevance in conveying the role of ginsenoside Rg5 in modulating the tumor microenvironment.
11. References
    Comment: Ensure citations are accurate and complete.
    Response: Thank you for the acknowledgment. We have cross-checked all references and updated citation formatting to ensure accuracy and completeness in the final manuscript.

 

 

Comments 2: Strengths of the Paper:

• Comprehensive Coverage:The review thoroughly covers a wide array of phytochemicals, their sources, molecular targets, and mechanisms in the context of breast cancer.
• Mechanistic Insights:It delves into critical signaling pathways (e.g., PI3K/Akt, MAPK, Wnt, TGF-β, Notch, VEGF, HER2) and cellular processes (apoptosis, cell cycle, angiogenesis, immune modulation).
• Illustrative Material:Including detailed figures and tables effectively summarizes complex information and enhances understanding. Figures 3, 4, and 5, detailing specific compound studies, are particularly informative.
• Relevance and Timeliness:The topic of natural compounds in cancer therapy is highly relevant and of growing interest.
• Discussion of Challenges:The authors appropriately address significant hurdles like bioavailability, standardization, and the need for robust clinical translation.
• Clear Structure:The manuscript is logically organized, making it easy for readers to follow the progression of ideas.

Response 2: We sincerely appreciate the reviewer’s thoughtful and encouraging remarks regarding the strengths of our manuscript. We are pleased to know that the comprehensive coverage of phytochemicals, their molecular targets, and the mechanistic insights into key signaling pathways have been well received. Our aim was to provide an integrative and up-to-date synthesis of current knowledge to benefit both researchers and clinicians exploring natural compounds for breast cancer therapy.

We are also grateful for the recognition of our efforts in presenting complex information through illustrative figures and structured tables. In particular, Figures 3, 4, and 5 were designed to visually reinforce the discussed mechanistic actions of specific phytochemicals, and we are glad they served their intended purpose effectively.

Moreover, we appreciate the acknowledgment of the manuscript’s structure and the discussion on translational challenges such as bioavailability and standardization. We believe that highlighting these hurdles is essential for guiding future research and facilitating the clinical applicability of natural compounds in oncology.

 

Author Response File: Author Response.pdf