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The authors have made the requested changes. However, the authors are advised to fix the typos and grammatical errors throughout the manuscript.
[# PeerJ Staff Note - this decision was reviewed and approved by Sonia Oliveira, a PeerJ Section Editor covering this Section #]
**PeerJ Staff Note:** Although the Academic and Section Editors are happy to accept your article as being scientifically sound, a final check of the manuscript shows that it would benefit from further English editing. Therefore, please identify necessary edits and address these while in proof stage.
Dear authors,
I hope this message finds you well, and appreciate the effort you and your team have put into the revisions. However, some concerns regarding the description of the methods and the presentation of results have been raised.
The description of the methods lacks clarity and it would greatly benefit the readers if there could be more detailed explanations to enhance comprehension. I believe a more thorough understanding of the methods employed would contribute significantly to the overall appreciation of your work.
Secondly, it seems difficult to analyze the results, the way they have been presented. The organization seemed a bit confusing, and the image quality, particularly when zoomed in, is not satisfactory. This hinders the ability to make meaningful interpretations. Perhaps providing higher-resolution images as well as reorganizing the results section could improve the overall readability and analysis of the findings. Additionally, pinpointing the controls used in your experiments does not appear to be easy. This may have been overlooked, but if it could be highlighted or reiterated more clearly, it would be immensely helpful.
Lastly, please provide a rationale behind the choice of cell lines as well as comment on their characterization, as the cells used are known to be highly mutable. It would be beneficial to elaborate on the selection of these elements or provide some context to help readers understand the significance of these choices in the context of your research. I
hope you find these comments constructive, and I am confident that addressing these concerns would greatly enhance the overall quality of your manuscript. I appreciate your time and effort in considering these suggestions. Thank you for your attention.
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Thank you for the revision of the manuscript titled "Circular RNA circMAN1A2 promotes ovarian cancer progression through the microRNA-135a-3p/IL1RAP/TAK1 pathway"
The reviewers have raised concerns regarding the provision of patient sample IHC data concerning the clinicopathological characteristics of circMAN1A2 in ovarian cancer patients. This crucial information is currently missing from the manuscript, which hinders the comprehensive understanding of the study's clinical implications.
Furthermore, there appears to be inconsistency in the reported number of mice used in the experiments. The manuscript initially stated that 20 mice were randomly assigned to 8 mice per group (n=8), but later, in figure 9, it indicates the use of 5 mice per group. This discrepancy creates confusion and needs to be addressed for the study to be reproducible and reliable.
To improve the manuscript, the authors must prioritize providing the necessary IHC data for patient samples, as this information is fundamental to understanding the clinical relevance of circMAN1A2 in ovarian cancer. Additionally, the authors should thoroughly review and rectify any discrepancies in the reported number of mice used in the experiments to ensure accuracy and consistency in the results.
This "final" revision is critical for the manuscript to meet the necessary standards for publication.
**PeerJ Staff Note:** Please ensure that all review, editorial, and staff comments are addressed in a response letter and any edits or clarifications mentioned in the letter are also inserted into the revised manuscript where appropriate.
The reviewer did not get any meaningful answer to each and every prior comments. The authors should elaborate their answer.
"A total of 20 mice were randomly assigned to 8 mice per group (n=8)."
What does that mean? The reviewer also found that 5 mice per group were used (figure 9). Surprisingly, the authors claimed that they changed animals/group into 8. This is very confusing. Is the reviewer missing something here?
Some figures (like cellular wound healing assay) are still very poor in quality.
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The articles fail to meet providing the patient sample IHC data for clinicopathological characteristics of circMAN1A2 in ovarian cancer patients.
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Li et al. in this new study focused on ovarian cancer (OC) and discovered that a potential circular RNA (circRNA) called circMAN1A2 may serve as an early diagnostic marker for OC. The authors investigated the effects of circMAN1A2 on OC cell proliferation using various experimental techniques. It was reported that circMAN1A2 was highly expressed in OC and played a role in promoting cell proliferation, clone formation, and tumorigenicity. Based on further analysis it is suggested that circMAN1A2 acts as a sponge for a specific microRNA (miR-135a-3p), which directly targets a protein called interleukin1 receptor accessory protein (IL1RAP). The authors report that the interaction between miR-135a-3p, IL1RAP, and circMAN1A2 ultimately affects the phosphorylation of transforming growth factor-beta-activated kinase 1 (TAK1), leading to increased OC cell growth. The findings suggest that circMAN1A2 may serve as a potential biomarker for the early detection of OC and a target for future therapeutic interventions.
The reviewers' questions should be carefully addressed, and the requested changes and improvements should be made in the manuscript before it is acceptable for publication in PeerJ.
[# PeerJ Staff Note: Please ensure that all review and editorial comments are addressed in a response letter and any edits or clarifications mentioned in the letter are also inserted into the revised manuscript where appropriate. #]
[# PeerJ Staff Note: The review process has identified that the English language must be improved. PeerJ can provide language editing services - please contact us at [email protected] for pricing (be sure to provide your manuscript number and title) #]
1. The use of different colors to differentiate between groups is welcome. However, the reviewer finds the colors to be little distracting and hence suggest the authors to use more sober colors whenever absolutely necessary.
2. There are several spelling mistakes: line no 180 -‘intraperitoneal injection of sodium pentobarbital (200 mg/kg body weight) and hotographed.’ in the manuscript.
3. The manuscript should also be checked by a fluent English speaker.
1. Xenograft studies in nude mice showed significant role of circMAN1A2 in OC. Why does circular RNA show this kind of marked effect? Is there any other example? Discuss in the manuscript please.
2. N=5 per group is not ideal for any animal experiment. Please repeat with increased N.
3. How many times did the authors replicate their data?
1. In wound healing experiments: why are the images in different color shades?
2. This is also true for clone formation assays.
3. The EdU staining: the reviewer could not find any significant difference between different groups in various cells, still the data was shown as significant. Please explain.
1. The authors should at least discuss the importance of other RNAs that may regulate the OC pathophysiology
The article “Circular RNA circMAN1A2 promotes ovarian cancer progression through the microRNA-135a-3p/IL1RAP/TAK1 pathway’’ is an interesting study detecting the specific mechanism of circMAN1A2 on ovarian cancer progression. However, the existing data are partially supportive of the authors' conclusions. Additional experiments are needed to fully support their findings. Major revision is needed in recommending this manuscript for publication.
1. The resolutions of figures specifically Fig. 2G, 4D, 5D, 7F, 8F. These figures need to be replaced with high resolution images.
2. Many figures are missing from this manuscript which were mentioned throughout result section Fig. 2H, 4E, 5E, 7H, 8H
3. The authors should determine the clinicopathological characteristics of circMAN1A2 in ovarian cancer patients.
4. The introduction should be rewritten for clarity.
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