Search Results (1,571)

Vascular calcification (VC) is a major complication of chronic kidney disease (CKD) that is strongly associated with cardiovascular mortality. While indoxyl sulfate (IS), a protein-bound uremic toxin, has been implicated in the progression of VC, the underlying molecular mechanisms remain unclear. We investigated the procalcific effects of IS using a two-step nephrectomy-induced CKD mouse model and cultured vascular smooth muscle cells. In vivo, progressive renal impairment was associated with elevated circulating IS levels and enhanced VC. In vitro, IS dose- and time-dependently induced calcium deposition, increased reactive oxygen species (ROS) production, and upregulated osteogenic markers, including runt-related transcription factor 2 (RUNX2), bone morphogenetic protein 2 (BMP2), and osteocalcin (OCN), whereas N-acetyl-L-cysteine (NAC) partially attenuated IS-induced ROS accumulation and cell injury. Mechanistically, IS exposure activated extracellular signal-regulated kinase (ERK) and Wnt/β-catenin signaling while suppressing nuclear factor erythroid 2-related factor 2 (Nrf2)-related antioxidant responses, as reflected by reductions in the phosphorylated Nrf2 (pNrf2) to total Nrf2 and heme oxygenase-1 (HO-1) expression. Pharmacological inhibition of ERK and Wnt/β-catenin signaling attenuated IS-induced osteogenic responses. These findings indicate that IS promotes VC in association with increased oxidative stress, activation of ERK and Wnt/β-catenin signaling, and impaired Nrf2-related antioxidant defense. These integrated findings provide mechanistic insight into IS-associated VC and highlight oxidative stress-related signaling networks as potential therapeutic targets in CKD. Full article

The corneal epithelium serves as the outermost transparent barrier of the eye and depends on rapid and coordinated cellular responses for wound repair. Although negative pressure (NP) has been shown to accelerate wound healing in other tissues, its cellular and molecular effects on corneal epithelium remain undefined. This study investigates how NP regulates corneal epithelial cell physiology and identifies the molecular mechanisms underlying NP-induced migration. Human corneal epithelial cells were exposed to normal or NP conditions, and cell motility was quantified using scratch-wound and transwell migration assays. Nuclear and cytoplasmic fractions were isolated for proteomic profiling to identify NP-responsive proteins. G3BP1 was selected as a candidate regulator and subsequently examined using molecular, biochemical, and functional assays to determine its role in NP-mediated signaling. Proteomic analysis revealed a significant NP-induced upregulation of G3BP1. Mechanistically, G3BP1 suppressed epithelial junctional proteins, including E-cadherin, p120-catenin, and ZO-1, while activating key pro-migratory signaling pathways involving AKT, ERK1/2, FAK, and Paxillin. These coordinated changes enhanced cytoskeletal dynamics and promoted corneal epithelial cell migration under NP stimulation. G3BP1 functions as a critical mechanotransduction mediator of NP, orchestrating adhesion remodeling and activating pro-migratory signaling cascades to facilitate corneal epithelial cell motility. These findings reveal a previously unrecognized cellular mechanism through which NP promotes epithelial repair and highlight G3BP1 as a potential therapeutic target for persistent corneal epithelial defects. Full article

Background: Atopic dermatitis (AD) is a persistent and recurring skin disease characterized by epidermal barrier dysfunction, immune dysregulation, and elevated expression of proinflammatory mediators. We investigated the anti-inflammatory potential of polynucleotides (PN), highly purified DNA biopolymers isolated from salmonid gonads, in keratinocyte and macrophage activation models. Methods: RAW 264.7 macrophages were stimulated with lipopolysaccharide (LPS), whereas HaCaT keratinocytes were stimulated with tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ). The effects of PN treatment on the production or expression of inflammatory mediators, cytokines, and chemokines were evaluated. Changes in the phosphorylation of mitogen-activated protein kinases (MAPKs) and Janus kinase 1/signal transducer and activator of transcription 3 (JAK1/STAT3) and in the nuclear localization of nuclear factor-κB (NF-κB) were also assessed. Results: In LPS-activated RAW 264.7 macrophages, PN treatment significantly suppressed nitric oxide production and downregulated the expression of inducible nitric oxide synthase (iNOS), TNF-α, IL-1β, and IL-8, accompanied by reduced NF-κB nuclear translocation. In TNF-α/IFN-γ-stimulated HaCaT keratinocytes, PN treatment markedly decreased the secretion levels of IL-6, IL-1β, and thymic stromal lymphopoietin. Moreover, PN treatment markedly reduced T-cell-recruiting chemokines, including MDC/CCL22, TARC/CCL17, RANTES/CCL5, and IL-8. Signaling analyses demonstrated that PN treatment attenuated the phosphorylation of key MAPKs (ERK, JNK, and p38) and the JAK1/STAT3 axis. Furthermore, PN treatment markedly reduced NF-κB nuclear translocation. Conclusions: These in vitro findings indicate that the anti-inflammatory effects of PN are associated with reduced activation of multiple core signaling pathways governing cytokine and chemokine responses, supporting further investigation of PN in AD and other chronic inflammatory skin diseases. Full article

The application of natural bioactive compounds in cosmeceutical products, particularly as skin-lightening agents, has received increasing interest. Caulerpa racemosa, a green macroalga of the Chlorophyta division, contains beneficial nutrients that are applicable as food and cosmeceutical ingredients. This study investigated the in vitro effect of the ethanolic extract of C. racemosa (CR) on regulation of melanogenic-related signaling and gene expression in SK-MEL-5 human melanoma-derived cells. Identification of bioactive components revealed that catechin, rutin, and quercetin as flavonoid contents were found in CR extract, analyzed using HPLC. The expressions of microphthalmia-associated transcription factor (MITF), extracellular signal-regulated kinase (ERK) signaling molecules, and melanogenic-related molecules were analyzed via Western blotting and qPCR. The CR extract treatment applied to SK-MEL-5 cells decreased the MITF protein expression level, which correlated with increased pERK expression, and no cytotoxic effect was observed. The subsequent treatment reduced the expression of melanogenesis-related genes (TYR, TYRP1, MC1R, and DCT) that were downstream targets of MITF. This study provides preliminary evidence that CR extract may modulate melanogenesis-related signaling. However, the specific bioactive compounds responsible for the observed effects remain to be identified, as the extract contains a complex mixture of phytochemicals. Further fractionation studies are needed to pinpoint the active constituents. The variability of extract composition due to seasonal and geographical factors should be considered for future standardization. Full article

Background/Objectives: Platelets are anucleate cells that play a crucial role in primary hemostasis and arterial thrombosis, contributing to cardiovascular diseases. Eugenol, a bioactive phenolic compound, exhibits vasodilatory, antibacterial, and anticancer properties and inhibits platelet aggregation induced by collagen and arachidonic acid (AA). AA is a critical lipid component of the platelet membrane and a precursor for potent signaling molecules that mediate platelet activation. However, the precise mechanisms through which eugenol modulates AA-stimulated platelet activation remain unclear. Methods: Human platelets were pretreated with eugenol and subsequently stimulated with AA. Platelet aggregation, ATP release, intracellular calcium mobilization, and P-selectin expression were measured. JNK, p38 MAPK, ERK, and vasodilator-stimulated phosphoprotein (VASP) phosphorylation were analyzed by Western blotting. SP600125, SQ22536, and ODQ were used to examine the involvement of JNK and cyclic nucleotide signaling pathways. Antithrombotic effects were further evaluated in a mouse mesenteric thrombosis model. Results: Eugenol significantly suppressed AA-induced platelet aggregation, ATP release, calcium mobilization, and P-selectin expression, selectively reducing JNK phosphorylation without affecting p38 MAPK or ERK. SP600125 produced similar inhibitory effects. Neither SQ22536 nor ODQ reversed eugenol’s inhibitory effects on platelet aggregation. Furthermore, eugenol did not alter VASP phosphorylation at Ser157 or Ser239. In vivo, real-time intravital imaging showed that eugenol and SP600125 delayed thrombus formation and prolonged occlusion time. Conclusions: These findings suggest that eugenol exerts inhibitory effects that may involve modulation of JNK phosphorylation, independent of cyclic nucleotide/VASP pathways in AA-induced platelet activation, highlighting its potential as an antithrombotic agent. Full article

Excessive osteoclast formation is a key contributor to pathological bone loss in disorders such as osteoporosis and rheumatoid arthritis. Asarinin, a natural lignan, has not previously been examined in the context of osteoclast differentiation. Here, we investigated the anti-osteoclastogenic effects of asarinin using RANKL-stimulated RAW264.7 cells. Asarinin significantly suppressed TRAP-positive multinucleated osteoclast formation. Mechanistically, asarinin selectively inhibited RANKL-induced phosphorylation of p38 and ERK MAPKs, leading to reduced c-Fos expression and inhibition of NFATc1 activation. In addition, asarinin disrupted actin ring formation in mature osteoclasts. Collectively, these findings identify asarinin as a pathway-selective inhibitor of osteoclast differentiation, acting through a mechanism consistent with modulation of the p38/ERK–c-Fos–NFATc1 signaling cascade while sparing parallel signaling pathways. Full article

In this study, we report for the first time the cloning and identification of three melatonin receptors (PgMtnr1a, PgMtnr1b, and PgMtnr1c) from the freeze-resistant fish Perccottus glenii, all of which encode typical GPCR proteins. These receptors are expressed widely in high-metabolism tissues such as the liver. During the freezing and recovery process at −2 °C, the expression of liver receptors exhibited dynamic changes: PgMtnr1a and PgMtnr1c were significantly upregulated during the middle of resuscitation, while PgMTNR1A reached its peak expression in the later stages, suggesting its involvement in stress repair. In vitro experiments confirmed that melatonin significantly enhances the activity of liver antioxidant enzymes in a receptor-dependent manner, an effect that can be inhibited by the antagonist luzindole. Anti-inflammatory analyses indicate that melatonin primarily inhibits inflammation-related genes through Mtnr1a and Mtnr1b. Furthermore, the overexpression of PgMTNR1A can synergistically activate ERK in the presence of melatonin, inhibit the JNK/p38 MAPK pathway, and downregulate the expression of NF-κB and COX2. Pathway inhibition experiments further validated that the MAPK/NF-κB axis, including ERK, JNK, and p38 pathways, mediates the anti-inflammatory effects of melatonin. This study systematically elucidates the molecular mechanisms by which the melatonin receptors of P. glenii play crucial antioxidant and anti-inflammatory roles during the later stages of freeze–thaw resuscitation, particularly by regulating the MAPK/NF-κB signaling axis through MTNR1A, thereby providing a novel basis for understanding the adaptation of vertebrates to extreme environments. Full article

Background/Objectives: Adenomyosis is a chronic gynecological disorder characterized by the presence of endometrial tissue within the myometrium, causing pelvic pain, abnormal uterine bleeding, and infertility. Despite its high prevalence, the molecular mechanisms underlying disease initiation and progression remain incompletely understood. Current evidence implicates disruption of the endometrial–myometrial interface, epithelial–mesenchymal transition, and progesterone resistance in driving tissue invasion and remodeling. Diagnosis relies mainly on imaging modalities, while reliable non-invasive biomarkers are lacking. MicroRNAs, as stable post-transcriptional regulators of gene expression, have emerged as key modulators of proliferation, inflammation, and hormonal signaling, and represent promising candidates for novel diagnostic strategies. Methods: A systematic review was conducted in accordance with PRISMA guidelines and registered with PROSPERO (CRD42025637752). A comprehensive search of the Medline, Embase, Scopus, and Cochrane databases was performed in April 2025. Studies investigating miRNA expression in patients with adenomyosis compared with controls were included. The quality of the studies and the risk of bias were assessed using the Newcastle–Ottawa scale. Two reviewers independently performed study selection, data extraction, and quality assessment. Results: Twenty-seven studies published between 2015 and 2025 met the inclusion criteria. Thirty-nine distinct miRNAs were reported as significantly dysregulated in adenomyosis. Recurrently altered miRNAs included let-7a, miR-145, miR-10b, miR-30c-5p, miR-141-3p, miR-143, and miR-191. Functional analyses have consistently implicated miRNAs in key pathogenic pathways, including Hippo-YAP, PI3K/AKT, MAPK/ERK, JAK/STAT, and Wnt/β-catenin signaling. These alterations were associated with enhanced epithelial–mesenchymal transition, increased cellular proliferation and migration, progesterone resistance, chronic inflammation, and immune modulation. Emerging evidence highlights exosomal and circulating miRNAs as promising non-invasive biomarkers, with a few studies already demonstrating diagnostic potential using serum, plasma, or urine samples. However, substantial heterogeneity in tissue types, sampling timing, and analytical methods precluded meta-analysis. Conclusions: MiRNAs play a central role in the molecular pathogenesis of adenomyosis and show strong potential as non-invasive diagnostic biomarkers. However, large-scale validation studies and standardized methodologies are required before clinical implementation. Full article

Objectives: Renal ischemia–reperfusion (I/R) injury is one of the most important pathological triggers of acute kidney injury. This study aimed to investigate the protective effects of meloxicam, a selective cyclooxygenase-2 (COX-2) inhibitor, against renal I/R injury through specific cell death pathways including inflammation, apoptosis, necroptosis, and the MAPK/ERK pathway, which is potentially linked to regulated cell death mechanisms such as ferroptosis. Methods: Male Wistar Albino rats weighing 280–300 g were used in the study and were divided into four groups: Sham, IR (40 min ischemia + 120 min reperfusion), Meloxicam + IR, and Meloxicam + IR1. Bilateral renal ischemia was induced for 40 min via a retroperitoneal approach under anesthesia. Meloxicam was administered intravenously at a dose of 1 mg/kg at the initiation of reperfusion in the Meloxicam + IR group, whereas in the Meloxicam + IR1 group, the same dose was administered 1 h after the onset of reperfusion. Total reperfusion time was 120 min in both groups. Renal function parameters (BUN and creatinine) and oxidative stress markers (TAS and TOS) were measured. Inflammatory cytokines (IL-6, IL-1β, and IL-10), the glomerular filtration injury marker Cystatin C, the tubular injury marker KIM-1, the apoptotic marker Caspase 3, the necroptosis markers RIPK3 and MLKL, and MAPK signaling pathway alterations (ERK1/2 and pERK1/2 levels) associated with cellular survival and death signaling were evaluated. Results: Most notably, meloxicam markedly modulated apoptosis, the expression of necroptosis markers RIPK3 and MLKL, and the activation of pERK1/2, a key node in MAPK signaling that is regulatory in cell survival and cell death processes. The drug also suppressed pro-inflammatory cytokines (IL-6 and IL-1β) while preserving anti-inflammatory IL-10 levels. Furthermore, improvements were observed in the levels of KIM-1, a marker of tubular injury, and Cystatin C, a marker of glomerular filtration impairment. Consequently, meloxicam administration significantly reduced the elevated serum creatinine and TOS levels observed in the IR group, although serum BUN levels remained without notable alteration. Conclusions: The findings of this study suggest that meloxicam may extend beyond its role as a classical anti-inflammatory agent, potentially offering biochemical and functional protection against renal I/R injury in association with the modulation of specific cell death mechanisms, including necroptosis and apoptosis, as well as the MAPK signaling pathway. Full article

Background/Objectives: Covalent inhibitors often demonstrate prolonged pharmacological effects even after their disappearance from the site of action because target recovery depends on target turnover. This disconnect between pharmacokinetics (PK) and pharmacodynamics (PD) complicates the development of such inhibitors since plasma exposure coverage of in vitro potency cannot be used for compound selection and human dose projections. In this study, we describe the development of a PK/PD modeling framework for covalent inhibitors. Methods: The model was developed for KRAS G12C inhibitors using pre-clinical data on sotorasib. The model was validated using data from both internal Amgen compounds and published data for several KRAS G12C inhibitors. The applicability of the framework was extended to EGFR covalent inhibitors by incorporating PK/PD and the efficacy of osimertinib and its active metabolite AZ5104. Results: The model successfully captured the pharmacokinetics, KRAS G12C target occupancy, inhibition of phosphorylation of ERK protein, and tumor growth inhibition following the administration of sotorasib in mice bearing MIA PaCa-2 xenografts. External validation with several internal Amgen compounds as well as publicly available data on KRAS G12C inhibitors showed the robustness of the model. The application of this framework to EGFR inhibitor Osimertinib and AZ5104 captured p-EGFR dynamics and resultant tumor growth inhibition. Conclusions: A modeling framework for covalent inhibitors was developed that links exposure to target occupancy, downstream signaling, and tumor efficacy. This framework could be useful for compound optimization and human dose projections. Full article

Chronic constipation is a prevalent functional gastrointestinal disorder with limited long-term treatment options. This study examined whether a fucoidan preparation derived from Sargassum fusiforme (SF) ameliorates loperamide (LOP)-induced constipation in male ICR mice. SF was administered by oral gavage at 50, 100 or 200 mg/kg/day for 35 days, and constipation was induced with LOP (5 mg/kg) during the final 7 days; the design is therefore preventive rather than therapeutic. Defecation endpoints were recorded at the cage level (n = 3 cages per group) and fecal moisture in individual mice (n = 7 per group), whereas colonic cytokine and myeloperoxidase (MPO) measurements (n = 3–4 per group) and Western blot analyses (n = 3 per group) were performed in a small subset of animals, and histological analyses in n = 6–8 per group. SF increased fecal output and fecal moisture content with increasing dose; at 200 mg/kg the relative fecal number returned to the level of the Vehicle control group, although the absolute daily pellet count remained lower than that of the Vehicle control and of the Positive control group. SF administration was associated with lower colonic TNF-α, IL-6, IL-1β and MPO concentrations, with reduced phosphorylation of p38, JNK and ERK, with higher C-kit and stem cell factor (SCF) levels, with normalization of the LOP-induced increase in aquaporin-3 (AQP3), and with recovery of colonic mucosal thickness and mucin content. 16S rRNA gene sequencing showed a shift in fecal microbiota community structure (PERMANOVA p = 0.0002) and a dose-related increase in the combined relative abundance of Lactobacillus and Bifidobacterium, without a reduction in alpha diversity. These findings indicate that SF ameliorates LOP-induced constipation in male mice and that this effect is accompanied by changes in colonic inflammatory signaling, motility-related protein expression, mucosal architecture and gut microbiota composition. Full article

Acute liver failure (ALF) is a fulminant hepatic syndrome characterized by rapid hepatocellular destruction, severe impairment of liver function, and high mortality. Effective pharmacological interventions capable of limiting early hepatic injury remain lacking. Upadacitinib (UPA), a selective inhibitor for Janus kinase 1 (JAK1) with established anti-inflammatory activity, has not previously been investigated in experimental ALF. Consequently, the current study examined the hepatoprotective potential and underlying mechanisms of UPA in a lipopolysaccharide (LPS)/D-galactosamine (D-GalN)-induced ALF murine model. Mice were pretreated with UPA (10 or 20 mg/kg) prior to LPS/D-GalN challenge. UPA significantly attenuated liver injury, as demonstrated by marked reductions in serum ALT, AST, ALP, and γ-GT levels, together with substantial improvement in hepatic histopathology, attenuation of necroinflammation, and reduction in neutrophil accumulation. UPA also restored hepatic redox balance through reduction in lipid peroxidation and nitrosative stress, alongside enhancement of antioxidant capacity. Mechanistically, UPA suppressed IFN-γ/JAK1/STAT1 signaling and downregulated NF-κB p65 and inducible nitric oxide synthase (iNOS) expression, with subsequent reduction in hepatic TNF-α production. In parallel, UPA inhibited MAPK pathway activation, including ERK1/2, JNK, and p38 signaling. Moreover, UPA attenuated hepatocellular apoptosis through suppression of active caspase-3 and Bax expression with restoration of Bcl-2 levels. The 20 mg/kg dose consistently produced greater biochemical, molecular, and histopathological protection than the lower dose. In conclusion, UPA confers significant protection against LPS/D-GalN-induced ALF through coordinated suppression of oxidative stress, inflammatory signaling, and apoptosis, primarily associated with inhibition of the IFN-γ/JAK1/STAT1 and TNF-α/NF-κB/MAPK pathways and modulation of the Bax/Bcl-2 ratio, underscoring its viability as a promising therapeutic candidate for ALF. Full article

Aortic dissection (AD) is a severe vascular condition marked by abrupt onset, rapid progression, and heightened mortality rates. Vascular smooth muscle cells (VSMCs), the predominant cellular component of the arterial media, are essential for maintaining the structural integrity and functionality of blood vessels. Recent studies have associated Cysteine-rich protein 2 (CSRP2) with the advancement of several vascular diseases. The involvement of CSRP2 in AD progression is unclear. Aortic tissues were collected from patients for RNA sequencing and histological analysis. A mouse model of AD was created using β-aminopropionitrile monofumarate (BAPN), while VSMC phenotypic switching was induced by platelet-derived growth factor BB (PDGF-BB). Adeno-associated virus vector was used to overexpress CSRP2 in aorta. A variety of histopathological assays and biochemical analyses were applied to determine gene and protein expression patterns as well as uncover underlying molecular mechanisms. CSRP2 was significantly downregulated in both human and murine AD, and CSRP2 gene overexpression dramatically reduced BAPN-induced AD incidence and prevented animal mortality. CSRP2 could preserve a contractile VSMC phenotype, even though under PDGF-BB stimulation. Mechanistically, our findings reveal that CSRP2 directly interacts with p130 Crk-associated substrate (p130Cas; also known as BCAR1) and reduces its phosphorylation, which in turn inhibits the activation of extracellular signal-regulated kinase (ERK) signaling pathways, thereby preventing VSMC phenotypic switching induced by PDGF-BB. Our findings identify CSRP2 as a novel regulator of VSMC phenotypic modulation and a significant modulator of AD development, suggesting its potential as a target for early intervention for AD. Full article

Ultra-processed foods expose consumers to heat-induced contaminants like 5-hydroxymethylfurfural (5-HMF), which impairs intestinal homeostasis via oxidative stress and inflammation. This study quantified 5-HMF in bakery products and evaluated blueberry polyphenols’ capacity to attenuate 5-HMF-induced cellular stress in an in vitro Caco-2 intestinal model. 5-HMF levels in commercial bakery products were determined using a colorimetric assay and HPLC-UV. Differentiated Caco-2 cells were exposed to 5-HMF (0.4 mM) for 48 h, alone or co-treated with blueberry juice (15 µg/mL of phenol equivalents). ROS production, lipid accumulation, gene expression of antioxidant, lipid, and inflammatory markers, and NF-κB/ERK signaling pathways were analyzed. HPLC-UV accurately quantified 5-HMF in bakery products (exceeding 2 mg/100 g), revealing systematic overestimation by the colorimetric method. In Caco-2 cells, 5-HMF significantly increased ROS, lipid accumulation, inflammatory cytokines (NLRP3, IL1B, and IL18), and activated NF-κB and p-ERK. Co-treatment with blueberry juice reduced ROS and lipids, upregulated the NRF2 antioxidant pathway, and drastically suppressed NF-κB -mediated inflammation. 5-HMF induced markers of metabolic dysfunction, oxidative stress, and inflammation in the intestinal epithelial model. Polyphenol-rich blueberry juice attenuated 5-HMF-induced alterations at the tested concentration, suggesting a potential protective nutritional strategy against epithelial stress associated with food-processing contaminants. Full article

Background/Objectives: Extra virgin olive oil (EVOO), a key component of the Mediterranean diet, has attracted research interest because olive-derived phenolics demonstrate potential anticancer activity in experimental models. This review summarizes evidence concerning whole EVOO, phenolic-enriched EVOO, olive phenolic extracts, and the isolated compounds hydroxytyrosol, oleuropein, oleocanthal, and oleacein. Methods: A structured narrative search of PubMed, Web of Science, ScienceDirect, and Google Scholar was conducted for literature published between 2015 and 2025. Evidence was reviewed for breast, prostate, colorectal, pancreatic, bone, oral, liver, gastric, hematological, and brain cancers. Comparatively limited evidence concerning cervical, endometrial, ovarian, melanoma, non-melanoma skin, and thyroid cancers was summarized separately. Results: The molecular evidence was derived primarily from cell culture and animal studies using isolated phenolics and concentrated extracts. Preclinical studies indicate that EVOO phenolics may demonstrate anticancer activity through multiple mechanisms, including antioxidant activity, anti-inflammatory effects, cell cycle arrest, induction of apoptosis, inhibition of metastasis, anti-angiogenic activity, and modulation of key signaling pathways, such as PI3K/AKT/mTOR, MAPK/ERK, NF-κB, JAK/STAT, Wnt/β-catenin, p53, and epithelial–mesenchymal transition-related pathways. Most molecular and pathway-level evidence was obtained using isolated phenolic compounds in cell culture or animal models, whereas evidence directly examining whole EVOO consumption was largely observational and substantially more limited. Experimental studies also reported that oleocanthal induced lysosomal membrane permeabilization, whereas hydroxytyrosol and oleuropein promoted mitochondria-mediated apoptosis. Furthermore, preclinical combination studies suggested enhanced tumor-cell sensitivity to selected chemotherapeutic, targeted, and immunotherapeutic agents. However, these effects have not been established in patients. Human evidence remains limited mainly to observational dietary associations and small exploratory interventions, with no conclusive demonstration of cancer prevention or therapeutic efficacy. Conclusions: Isolated EVOO-derived phenolic compounds demonstrated promising anticancer mechanisms in preclinical models. However, these results should not be directly extrapolated to dietary EVOO because experimentally administered doses, bioavailability, metabolism, and food-matrix interactions differ substantially from human dietary exposure. Therefore, well-designed studies using chemically characterized EVOO, pharmacokinetic investigations, and controlled human trials are required before dietary or clinical recommendations can be made. Full article