Long non-coding RNA LINC00467 regulates hepatocellular carcinoma progression by modulating miR-9-5p/PPARA expression

The aim of this study was to analyse the expression pattern and elucidate the mechanistic involvement of long non-coding RNA LINC00467 in hepatocellular carcinoma (HCC). The relative expression of LINC00467 and microRNA (miR)-9-5p was determined by real-time polymerase chain reaction. Cell viability was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The cell proliferation was analysed by cell counting. Cell migration and invasion were monitored by Transwell assay. The luciferase reporter assay was employed to investigate the regulatory effect of miR-9-5p on LINC00467 and peroxisome proliferator-activated receptor alpha (PPARA). The endogenous PPARA protein was quantified by western blotting. It was found that LINC00467 was aberrantly decreased in HCC. The ectopic expression of LINC00467 significantly suppressed cell viability, proliferation, migration and invasion. LINC00467 functioned as a sponge for miR-9-5a and negatively regulated miR-9-5p expression. We also identified PPARA as the direct target of miR-9-5p. siRNA-mediated knockdown of PPARA in LINC00467-proficient cells promoted cell viability, migration and invasion. Our data indicate the critical involvement of LINC00467/miR-9-5p/PPARA signalling in the incidence and progression of HCC.


Introduction Yes
Is the length of the paper justified? Yes Should the paper be seen by a specialist statistical reviewer? No

Do you have any ethical concerns with this paper? No
Comments to the Author In this MS entitled "Long noncoding RNA LINC00467 regulate Hepatocellular Carcinoma progression by modulating miR-9-5p/PPARA expression", the authors did a nice work to elucidate the role of LINC00467 in the hepatocellular carcinoma. It is found that LINC00467 is aberrantly decreased in hepatocellular carcinoma, while ectopic expression of LINC00467 significantly suppresses cell proliferation, migration and invasion in two cell lines SMMC-7721 and HepG2. Furthermore, LINC00467 is found to function as sponge of miR-9-5a and negatively regulates miR-9-5p expression, which target is PPAR. These results are interesting and add new knowledge to the understanding of long noncoding RNAs in hepatocellular carcinoma. Although the findings deserve publication, some clarifications and modifications are needed. They are outlined as follows. 1--Abstract: Please unify the verb tense here. Sometimes it is passive tense, sometimes it is active tense, especially in presenting the results. 2--Introduction: In the last several sentences, please list your hypothesis or study design instead of results or findings. 3--Methods and materials: It seems that the data for figure 1 come from the clinical patients, but I can not find any information in the methods. 4--Results: The unit for Figure 2B Y axis is not correct. It will be better to change it. 5--Discussion: Some more in-depth discussion is needed. 6--Figure legends: In some figure legends, the statistical method is introduced, while there is not in other figures.

10-May-2019
Dear Dr Li, We are writing to inform you that the Editor has reached a decision on your manuscript RSOB-19-0074 entitled "Long noncoding RNA LINC00467 regulate Hepatocellular Carcinoma progression by modulating miR-9-5p/PPARA expression", submitted to Open Biology.
As you will see from the reviewers' comments below, there are a number of criticisms that prevent us from accepting your manuscript at this stage. The reviewers suggest, however, that a revised version could be acceptable, if you are able to address their concerns. If you think that you can deal satisfactorily with the reviewer's suggestions, we would be pleased to consider a revised manuscript.
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Reviewer(s)' Comments to Author(s):
Referee: 1 Comments to the Author(s) I thank the editor for the assignment of the review task of this manuscript, and I thank the authors for their contribution. This paper describes the involvment of long noncoding RNA Linc00467 in hepatocellular carcinoma progression and the underlying molecular mechanisms. They provided clinical and in vitro data to support the importance of Linc00467 in the hepatocellular carcinoma. Overall, the data looks nice and convincing. Some revions are preferred before publishing these results. The comments are specified below: Major concerns: 1. The number of samples for normal and cancer is not equal in Figure 1A. Please explain. Also, what did you mean "normal"? Did you mean healthy participants? 2. The idea to employ two cell lines is good. However, it would be nice to include some discussion of limitations about the findings in other cancer cells. Minor concerns: 3. The "Hepatocellular Carcinoma" in the title should not be captilized. 4. The first sentence in the abstract is not complete. Please use complete sentence here. 5. Some abbreviations, including in the title and other sections, are not defined. Please note that there is no need to abbreviate them if used only once. 6. Do you justify the cells before use? 7. Some more experimental details are required, such as the PCR primer sequence. 8. MTT is usually employed for cell viability assessment, not proliferation. Please check. 9. S.D is wrongly defined in the Statistical analysis. 10. It seems that some set of the data is not appropriate to be analyzed with ANOVA analysis. Please check. 11. Some typos should be corrected throughout the manuscript.

Referee: 2
Comments to the Author(s) In this MS entitled "Long noncoding RNA LINC00467 regulate Hepatocellular Carcinoma progression by modulating miR-9-5p/PPARA expression", the authors did a nice work to elucidate the role of LINC00467 in the hepatocellular carcinoma. It is found that LINC00467 is aberrantly decreased in hepatocellular carcinoma, while ectopic expression of LINC00467 significantly suppresses cell proliferation, migration and invasion in two cell lines SMMC-7721 and HepG2. Furthermore, LINC00467 is found to function as sponge of miR-9-5a and negatively regulates miR-9-5p expression, which target is PPAR. These results are interesting and add new knowledge to the understanding of long noncoding RNAs in hepatocellular carcinoma. Although the findings deserve publication, some clarifications and modifications are needed. They are outlined as follows. 1--Abstract: Please unify the verb tense here. Sometimes it is passive tense, sometimes it is active tense, especially in presenting the results. 2--Introduction: In the last several sentences, please list your hypothesis or study design instead of results or findings. 3--Methods and materials: It seems that the data for figure 1 come from the clinical patients, but I can not find any information in the methods. 4--Results: The unit for Figure 2B Y axis is not correct. It will be better to change it. 5--Discussion: Some more in-depth discussion is needed.

c) Introduction Yes
Is the length of the paper justified? Yes

Do you have any ethical concerns with this paper? No
Comments to the Author Good revisions. No further comments.

Accept as is
Are each of the following suitable for general readers?

c) Introduction Yes
Is the length of the paper justified? Yes

Do you have any ethical concerns with this paper? No
Comments to the Author No comments.

20-Jun-2019
Dear Dr Li We are pleased to inform you that your manuscript RSOB-19-0074.R1 entitled "Long noncoding RNA LINC00467 regulate hepatocellular carcinoma progression by modulating miR-9-5p/PPARA expression" has been re-reviewed and no further changes have been recommended by the referees.
The Editor is ready to accept the manuscript however, there is one remaining query regarding the Western blot films in Figures 5D and 5E that we would be grateful if you could address. When comparing the bands in the figures 5D and 5E with the raw files provided, the bands do not appear to match. We would be grateful if you could re-supply the raw blots with weight markers. The two PPARA panels also appear to have vertical splice lines as attached; please can you review this and provide the uncut files or an explanation for this.
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2) A separate electronic file of each figure (tiff, EPS or print-quality PDF preferred We are pleased to inform you that your manuscript entitled "Long noncoding RNA LINC00467 regulate hepatocellular carcinoma progression by modulating miR-9-5p/PPARA expression" has been accepted by the Editor for publication in Open Biology.
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