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All issues pointed out by the reviewers were adequately addressed and the revised manuscript is acceptable now.
[# PeerJ Staff Note - this decision was reviewed and approved by Sonia Oliveira, a PeerJ Section Editor covering this Section #]
No comments
The statistical analysis in supplementary fig S2, Fig S3 is missing
Improved very well
The authors have worked extensively to improve the MS. It can now be published just that authors should do a small correction by including statistical analysis in their new figures in supplementary sections.
Please address the concerns of all reviewers and revise the manuscript accordingly.
[# 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. #]
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Please find the attached file for the review comments.
In the manuscript, Anti-ENO1 antibody combined with metformin against tumor resistance: a novel antibody-based platform (#83201) the authors have shown that combinatorial treatment of Antibody along-with Metformin could reverse chemoresistance. This study also provides an interesting fact that heterogeneity in Cancer stem cells could be one of the causes of chemoresistance. I recommend the manuscript for publication subject to responses to the following comments.
1) Authors should consider rephrasing the sentence (from line 48 to 51) as it not very clear or try to split the sentence
2) The study would make more sense and would be easy to understand if authors could show the spheroid formation (Spheroidization in Fig.1.B) across days as mentioned in the methods section, as this may represent that spheroid formation was altered initially but after few days it become unchanged due to the self-renewal abilities of cancer stem cells (that would eventually lead to drug resistance).
3) Authors should also consider characterizing the Cancer stem cells by quantifying markers for stemness (like NANOG in normal vs spheroids via qPCR) as well as some chemoresistance genes.
4) In Fig.2 A and B. Authors have shown that DPP and CTX could upregulate ALDH+ and CD44+ CSCs subpopulation. It would be interesting if authors could also look at the expression of drug efflux transporters in these subpopulations of CSCs. Authors should also use a bigger font or try to represent it in a more legible manner.
no comment
5) Authors shall also consider putting the name of cell line in Figure.3A (Cell proliferation and IC50 experiment). Please try to split the figure to make it more legible.
The manuscript is well written. no comment.
Material and Methods need to be more detailed. for example, it is not clear how many technical replicates were considered in the cell proliferation, colony formation assay.
I doubt the novelty of the study.
Shu et al found that combined treatment of anti-Eno1 and MET could simultaneously target ALDH+ and CD44+ subpopulations and decrease the number of migration, invasion, colon and sphere cells, and proliferation potential, as well as overcome the drug resistance. The results are interesting and significant to the field, although the roles of Eno1 and MET on different cancer types are well-known. The novelty of the study is dramatically decreased by a published paper “ZHANG H, YANG T, YU Z, et al. Anti-ENO1 antibody combined with metformin reverses the resistance of human non-small cell lung cancer A549 cells to cetuximab by targeting cancer stem cells[J]. Chinese Journal of Cancer Biotherapy, 2021: 239-246.”, since they showed similar findings with combined treatment of anti-Eno1 and MET on cancer stem cells. Thus, I would not recommend this paper, unless the authors have further data showing underlying mechanisms by co-treatment of anti-Eno1 and MET or provide vivo data,
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