Isocorydine Exerts Anticancer Activity by Disrupting the Energy Metabolism and Filamentous Actin Structures of Oral Squamous Carcinoma Cells

Round 1

Reviewer 1 Report

Does it have little damage to normal cells (HGF) how it was proved? This part should be better described

Is ICD cancer specific drug? That means, does it target cancerous cells?

Conclusion are:

ICD can inhibit the proliferation of oral squamous cells and destroy their cytoskeletal structure. It also can induce mitochondrial dysfunction and energy metabolism disorder of cancer cells, and finally accelerate cell apoptosis. It showed good anticancer activity against OSCC, showing excellent anticancer activity against OSCC. 

What about normal cells and non-cancerous cells?

Minors editing 

Author Response

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Reviewer 2 Report

The paper of Zhou et al. “Isocorydine Exerts Anticancer Activity by Disrupting the Energy Metabolism and Filamentous Actin Structures of Oral Squamous Carcinoma Cells” is devoted to study of Isocirydine’s antitumor potential in oral squamous carcinoma cell models.

My comments:

1)    In line with CCK-8 assay, tests for migration and apoptosis, the authors should carry out the simple cell cycle analysis.

2)    Abstract: The results will be much more visual and impressive if the authors show the suppression of the activity of mitochondrial complexes as a percentage relative to the control.

3)    Introduction: “Moreover, ICD can also combine with first-line anticancer drugs such as gemcitabine [22], doxorubicin (DOX) [27] and cisplatin to enhance anti-cancer sensitivity and reduce drug resistance [26].”

Please, replace with “can be combined with.. ’

4)    Figure 1 Legend: not “migration effect” and “invasion effect”, but, for instance “anti-migration” and “anti-invasion effect”,

5)    2.3 Proteins  “ARP3”, :ARPC3” appear unexpectedly. Please, describe, why is it important to analyze them in this case?

What is “AFM?” Please, explain the term before you use it first time

6)    4.7. Cell mitochondrial function analysis

The authors should provide at least the catalog number of assay kit used. One can suggest that Mitotracker was used to assess MMP, whereas ROS level was analyzed by H2DCFDA staining. However, it should be noticed. And what about kits for the analysis of respiration complexes activities?

What was the kit used for ATP content quantification?

Minor editing of English language required

Author Response

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Reviewer 3 Report

Zhou et. al. showed that Isocorydine (ICD) has strong antitumor effects on OSCC. They showed that invasion, migration, and adhesion were decreased in Cal-27 cells after ICD treatment. They found that ROS production was increased while MMP expression and ATP content were decreased. They conclude that ICD has anticancer activity against OSCC. Their work is interesting and shows the effect of ICD on the cancer cell. 

1- the work is well described, however, one cell line, Cal-27, is not enough to draw conclusions. The authors should add one or two cell lines to confirm the findings of this project. 

2- Please write the % of apoptotic and dead cells in each quadrant in Figures 4E and 4F. 

3- On the Material and Methods at 4.2 anticancer activity: the authors wrote “media without drugs were added to the control group". Was DMSO added to the media for the controls?

Minor editing of English language required

Author Response

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Reviewer 4 Report

This study shows that ICD can potentially inhibit the proliferation of oral squamous cells and destroy their cytoskeletal structure. More importantly, ICD can induce apoptosis, mitochondrial dysfunction, and energy metabolism disorders in cancer cells. However, the study is interesting, and the authors should address the following comments:

1.    Do the authors examine the toxicity of ICD in normal oral squamous cells?

2.    Figure 2C. The western blot result and the graphical ARPC3 expression are not compatible. Describe.

3.    It would be beneficial for the paper if the authors included the results of the western blot for apoptosis.

4.    It would be promising if the authors conducted an animal experiment on this subject.       

Author Response

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Round 2

Reviewer 3 Report

The authors implemented the changes suggested by the reviewers and improved the manuscript.