The Dysregulation of MicroRNAs in the Development of Cervical Pre-Cancer—An Update

Round 1

Reviewer 1 Report

The current study explored the Dysregulation of MicroRNAs in the Development of Cervical Pre-cancer, and overall，this review is of great interest. At first, a large body of evidence shows that MicroRNAs are involved in multiple processes in the pathogenesis of cervical cancer; secondly, the review of the role of MiRNAs in cervical pre-cancer is not sufficient, and this review focuses on the dysregulated miRNAs in the cervical pre-cancer stage; moreover, the current review summarized Dysregulation of miRNAs in cervical pre-cancer, HPV induced miRNA changes in cervical pre-cancer, and the miRNAs with critical roles in early transformation of cervical cells. Thus, the phenotypes and mechanisms of the role of MicroRNAs in cervical pre-cancer have been described in detail.

Comments for authors:

1)        We recommend citing more of the authors' own research.

2)        The conclusion section needs to summarize more comprehensively the shortcomings of previous studies and the significance of this review, and present the authors' own views and directions for subsequent research

Author Response

Please refer to the doc.

Author Response File: Author Response.pdf

Reviewer 2 Report

The manuscript entitled " The dysregulation of MicroRNAs in the development of Pre-Cervical Cancer" is well written and informative. This kind of review article is much needed to improve the treatment strategies and a hot topic in cancer.  However there are some points that need to be improved: 1. Throughout the manuscript there are some red colored text, the authors should change it in uniformed manner. 2. In Abstract: Please include the abbreviation for HPV. 3. Line number 104 to 118, should be rephrased in para 2 line number 46.... 4.Please check the plagiarism percentage, some of the sentences are need to be rewritten. 4. Page 21, Line 588, HPV16 should be changed as HPV 16. 5. Include serial numbers in all the tables. Also, please mentioned the miRNA in orderly manner by year. 6. The authors should include some sentences about radiation induced change of Micro RNAs.    

Author Response

please refer to the doc.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

The authors carried out all the necessary corrections. Therefore I recommend this manuscript for publication.

This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.

Round 1

Reviewer 1 Report

None

Author Response

We thank reviewer 1 for his/her precious time spent reviewing our paper. As reviewer 1 did not leave any comments, we do not attach any response here.

Reviewer 2 Report

Choi et al. reviewed the literature and provided in the manuscript a summary of dysregulated miRNAs in cervical precancerous tissue. Two review articles on the same topic have summarized relevant literature published before 2017. In recent 5 years, some more studies have emerged in the field, and the new findings are shown to be in agreement with those summarized previously, to a great extent, as demonstrated in Table 1 in the current manuscript.  

The manuscript contains countless problems, some of which are as follows:

1. It is an update review, which should be reflected in the manuscript title;
2. The statement in the abstract that reads as “Currently, there is no review article which analyses the relationship between the dysregulated miRNAs in pre-cervical tissue and their possible roles in the early process of normal cell transformation” is inappropriate, as two review articles on the same topic have already been published;
3. In the first paragraph of the Introduction section, the authors miss the HPV vaccine as a significant contributor for the drop in cervical cancer incidence and death in the USA;
4. The content of lines 42-45 is repeated in lines 129-133. The authors can introduce the Bethesda system in the second paragraph of the Introduction section and delete lines 129-133;
5. Lines 57-63 on DDR and HPV replication are verbose;
6. What terms were used for literature search? What databases were searched? The information is missing;
7. The first paragraph in section 2.1 describes the detailed results of two previous review articles, which can be concise, as Table 1 already contains the same information;
8. In Table 1, whether a miRNA is upregulated or downregulated should be indicated. Is every miRNA upregulation (or downregulation) consistent across all studies? The answer to this question as a footnote would be helpful;
9. Zeng’s work has been summarized in a previous review article. The second paragraph in section 2.1.1 should be significantly shortened;
10. While citing Zeng’s study, the author write that miR-218 was found downregulated in both LSIL and HSIL tissue (lines 148-149), but then state that “the down regulation of miR-218 was not significant in pre-cervical cancer tissue” (lines 160-161). Please explain the contradicting depictions;
11. Lines 169-173 about “a progressive up-regulation of miR-21, miR-146a, and miR-182…” are verbose;
12. For lines 173-176 about let-7, please specify the two downregulated family members of let-7, and indicate loss of which family member(s) leads to the development of poorly differentiated, aggressive cancers;
13. Lines 173-174 describe Lee’s study as “two family members of let-7 were found down regulated throughout the cervical cancer progression”, but Lee et al. did not investigate let-7 in cervical cancer (ref 33);
14. The authors stress that Section 2 focuses on new studies conducted in recent 5 years (2017-2021) (line 95), but from the third paragraph in section 2.1.1 on, it becomes difficult to tell which studies reviewed are new;
15. The authors write in lines 243-249 that “When we further analyzed the miRNAs list in table 1, according to the qRT-PCR results, we found that when CIN1 was compared to the normal, the two most up-regulated miRNAs were miR-155-5p and miR-205-5p…”, but by viewing Table 1 in the manuscript, I cannot understand how the authors obtain the results. I guess “table 1” is from the cited study;
16. “cervical samples” in line 275 needs to be corrected;
17. Blood draws are considered minimally-invasive. Therefore, “non-invasive” should not be used for testing involving blood samples;
18. It seems that “and specificity” in line 384 should be removed;
19. “cellular” in line 393 should be deleted;
20. Table 2 is largely illegible;
21. Two “Table 3” are shown in the manuscript;
22. Lines 489 states that “The HPV genome contains three central regions”, but only two regions are introduced.
23. Citation of Figure 1 in the Conclusions section is inappropriate;
24. Figure 1 is messy: 1) in the column of “Normal Cervical Cells”, I do not understand how up- and down-regulated miRNAs are determined in normal blood and mucus samples, and why blood and mucus (they are not cells) and HPV-infected cell (not the normal cell) testing results are shown; 2) I do not understand why data without statistical significance are presented; and 3) are up- and down-regulations of miRNAs in CIN1-3 samples determined by comparing with those in normal tissue? Additionally, figure legends should be provided.    
25. “pre-cervical tissue” is used throughout the manuscript. Better change it to “cervical precancerous tissue”. Also, change “pre-cervical cancer” to “cervical pre-cancer”.
26. Grammatical errors: “the non-invasive pre-malignant lesions that subsequently transforms into invasive cervical cancer (lines 41-42); “E1-E2-mediate viral replication is essential for HPV genome amplification” (line 499); “The HPV-16 viral load and miR-139-3p expression in head and neck cancer tissue samples was negatively correlated” (lines 510-512); etc.

 

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

I have mentioned that there are countless issues in the manuscript and some of these have been listed in the last round of review, hoping the authors can identify and fix the rest. Unfortunately, I do not think the changes the authors have made constitute a significant improvement.

1) “HPV’s E1-E6 protein” in line 28 should be modified, as HPV produces E1, E2, E4, E5, E6 and E7 proteins (no E3).   

2) Accurate citation and proper interpretation of original studies are vital to a review article. The current manuscript contains numerous problems in reference citations. Some examples are as follows.

-Liu’s study (ref 6) is referenced several times (line 273, line 278, line 391, lines 484-486, and lines 510-515) but ref 6 is by Zubillaga-Guerrero et al. 

-Li et al. measured pri-miR-34a expression in normal cervical epithelium with (n = 32) or without (n = 32) HR-HPV infection, and cervical intraepithelial neoplasia (CIN) with (n = 32) or without (n = 12) HR-HPV infection, and found that pri-miR-34a was significantly lower in normal cervical epithelium and CIN with HR-HPV infection than in those without HR-HPV infection (ref 55). Why do the authors not present the finding with CIN that is more important? (lines 486-490).

-Lines 490-495 describe a study by Zamani et al. but miss key information: miR-21 was significantly up-regulated while miR-21 was significantly downregulated in patients with CIN or invasive cervical cancer than in the controls (HPV and cancer free).

-The authors cite a study by Lu and Gu in lines 586-594 and state that “As the author did not mention the status of the patients with HPV infection, our estimate is the patients were at the very early cervical pre-cancer stage and an immune response was triggered to fight against the HPV infection”. This statement is exaggerative and misleading, as in the original article only serum samples from patients with HPV 16 infection were tested, that identified miR-221 upregulation, while overexpression of miR-221 leading to increased IFN-α and IFN-β production was found in in vitro cell models.   

3) Figure 1

-Viral genome amplification is indicated only for HPV-infected cells, and I do not understand why so many miRNAs that are not found to be dysregulated in HPV-infected cells and their respective targets are illustrated in the “Genome Amplification” box. Here, better change “Genome Amplification”  to “HPV Genome Amplification”.

-In the column of “HPV-infected cells”, the dark-blue spheres, I guess, represent viral particles. The size of these spheres (relevant to the cell size) is inappropriate. A description of these spheres as figure legend would be helpful. I suggested adding figure legend in the last comments, that however is not seen in the revised manuscript.

-“Data from microarray is treated as not significant”. How does the reader know which data are from microarray analysis? Again, figure legend may help.

-MiR names shown in “Blood” are hardly legible.

-For columns CIN1-3, I do not understand why some miRNAs are listed at the top and some at the bottom.