NEAT1–SOD2 Axis Confers Sorafenib and Lenvatinib Resistance by Activating AKT in Liver Cancer Cell Lines
, Ririko Shinonaga 1,†, Hiromi Sakaguchi 2, Yutaka Kitagawa 2 and Kenji Yoshida 2
Round 1
Reviewer 1 Report
Current Issues in Molecular Biology
COMMENTS TO THE EDITORS AND THE AUTHORS
Manuscript ID cimb-2112574: “NEAT1-SOD2 axis confers sorafenib and lenvatinib resistance by activating AKT in hepatocellular carcinoma cells"
Dear the Editors and the Authors,
Please find below some of the comments for the above-mentioned manuscript.
SUMMARY OF THE CONTENT
The authors stated that their study investigated the effects of NEAT1v1 on drug resistance in human hepatocellular carcinoma cell (HCC) lines HLE and HuH6. Authors stated that results showed that NEAT1 knockdown activated MAPK signaling, including MEK/ERK, but suppressed AKT. The NEAT1 knockdown sensitized HCC cells to sorafenib and lenvatinib, but it conferred resistance to capivasertib. NEAT1v1 overexpression suppressed MEK/ERK and activated AKT, resulting in resistance to sorafenib and lenvatinib and sensitization to capivasertib. SOD2 knockdown reverted the effects of NEAT1v1 overexpression on the sensitivity to the molecular-targeted drugs. The authors concluded that NEAT1v1 switches the growth modality of HCC cells from MEK/ERK-dependent to AKT-dependent mode via SOD2 and regulates sensitivity to the molecular-targeted drugs independent of ER stress.
THE OVERALL OPINION OF THE MANUSCRIPT
The strengths: the manuscript presents new and interesting results; the manipulations with different signaling molecules were used to obtain the results; the mechanistic approach was used; the figures very clearly present the result.
The limitations: the title, the parts of the text as well as the conclusions are not precisely formulated since specific cell lines were used; the citation of the original, and important pioneered findings in the field focusing on the subject of the study are missing; the intra- and inter- assay coefficients are not provided; the limitation of the study (artificial in vitro system; only male sex of the cell lines) were not discussed and pointed.
(1) TITLE
Please consider modifying the title to precise state the type and sex of the cell lines used. Namely, the results are obtained using HLE cell line originated from 68-year male and HuH6 cell line originated from 1-year male. However, the title suggests general findings.
(2) ABSTRACT
Please consider modifying the text of the abstract by removing the introduction and by adding the limitation of the study.
(3) INTRODUCTION
3.1. Please describe the original and important pioneered findings in the field focusing on the subject of the study. Namely, the original and important pioneered findings were not cited.
(4) MATERIALS AND METHODS
4.1. Please provide the Key resources table.
4.2. Please provide the amount of RNA used for real-time PCR analysis.
4.3. Please provide the amount of the protein used for western blot analyses.
4.4. Please provide the Ct values for the genes discussed and please add the values in SuppTable1.
4.5. Please provide intra- as well as inter-assay coefficients for all analyses.
(5) RESULTS
Please be precise in describing/stating the results related to the specific cell line.
(6) DISCUSSION
6.1. Please discuss the original and important pioneered findings in the field focusing on the subject of the study.
6.2. Please discuss the limitations of the study (artificial in vitro system; only male sex of the cell lines).
(7) CONCLUSIONS
7.1. Please modify the text of the conclusions to precisely state the type and the sex of the cell lines.
(8) REFERENCES
8.1. Please provide references describing the original and important pioneered findings in the field focusing on the subject of the study.
I would greatly appreciate if you will contact me if you find something in my comments is missing/unclear/incorrect.
Good luck and all the best J
Author Response
Please see the attachment.Author Response File: 
Author Response.pdf
Reviewer 2 Report
Neat1 involvement in resistance to the therapy has been a major problem in treating hepatocellular carcinoma (HCC). In order to improve the therapy for HCC, it is very important to study the pathway and the key molecules that are activated or interact with this protein. In this study you have provided the experimental evidence to show that the absence of this protein makes the HCC cell lines sensitive to sorafenib and Lenvatinib, but resistance to capivasertib as phosphorylation of AKT is inhibited. Dissimilarly overexpression of NEATv1 causes HCC cell lines resistant to sorafenib and Lenvatinib, but sensitive to capivasertib. The evidence provided to emphasis the importance targeting the AKT is very commendable
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Reviewer 3 Report
The manuscript by Tsuchiya et al., might be interesting but need more work to be done. In terms of mechanism it seems average.
Major Comment : My major concern about the manuscript is that Authors could not show the major difference in cell viability assays in KD cells vs the controls, which can lead to the re-emergence of the tumor in invivo models.
Authors are suggested to reconsider the significance of the study as there is not a big difference in any of the cell viability assay comparisons.
Minor Comment:
1) Authors did not show any blots confirming the KD of NEAT1 including any supplementary figures.
2) Authors have showed the faded or highly brightened blots in figures, GAPDH fig 1A, Fig 2A, Fig 3A, 3C, 4B, 4C, 4E
b tub Fig 5B.
Therefore, authors are suggested to correct it to show the real blots without major contrast change or brightness change just to display the more original and reliable results.
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
