All reviews of published articles are made public. This includes manuscript files, peer review comments, author rebuttals and revised materials. Note: This was optional for articles submitted before 13 February 2023.
Peer reviewers are encouraged (but not required) to provide their names to the authors when submitting their peer review. If they agree to provide their name, then their personal profile page will reflect a public acknowledgment that they performed a review (even if the article is rejected). If the article is accepted, then reviewers who provided their name will be associated with the article itself.
Congratulations, your paper has been accepted.
[# PeerJ Staff Note - this decision was reviewed and approved by Vladimir Uversky, a PeerJ Section Editor covering this Section #]
Please revise the figures and resubmit at the earliest.
The length of the scale bars is not specified. Please update the figure/legends by adding the corresponding scale bar lengths. Then the manuscript will be in a good format and ready for publication.
no comment
no comment
Comments of the reviewers 1 and 3 were answered and their suggestions were incorporated in the modified manuscript.
The authors have followed well defined methods systematically and carried out the experiments.
Already mentioned with respect to the original manuscript.
Nil
Please revise and resubmit at the earliest.
**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.
This study explores the expression level of G6PD in lung tissue samples from 64 patients and its role in Non-Small Cell Lung Cancer (NSCLC) cell lines in the context of cell apoptosis. The analysis of clinical data is comprehensive and likely to stimulate substantial research interest. Additionally, the paper is well-structured, providing a smooth reading experience.
However, most of the findings only stem from in vitro cell line studies.
For a more compelling perspective, I recommend integrating ex vivo lung cancer tissues or an in vivo NSCLC mouse model. This would enhance the validity of G6PD as a therapeutic target, as evaluated using DHEA or D-(-)- ribose.
The following specific concerns need to be addressed.
The manuscript entitled "Inhibition of non-small cell lung cancer (NSCLC) proliferation through targeting Glucose 6-phosphate dehydrogenase{G6PD}described the growth-promoting effects of G6PD in non- small cell lung cancer (NSCLC)..
Clinical characteristics and G6PD expression levels in lung tissues of 64 patients
diagnosed with lung cancer at the King Chulalongkorn Memorial Hospital (Bangkok, Thailand)
during 2009-2014 were analyzed. G6PD activity in NSCLC cell lines, including NCI-H1975 and
NCI-H292, was experimentally inhibited using DHEA and siG6PD to study cancer cell proliferation and migration.
DHEA{Dehydroepiandrosterone}, an adrenal steroid is a potent uncompetitive inhibitor of mammalian glucose-6-phosphate dehydrogenase (G6PDH). Following methods have been used to meet the objective of the study.
{A.}Tissue specimens :-Lung tissue specimens were obtained from 64 lung cancer patients who were
admitted to the King Chulalongkorn Memorial Hospital (Bangkok, Thailand) during 2009-2014.
All tissue samples were examined by a team of pathologists at the King Chulalongkorn
Memorial Hospital to determine the type and stage of cancer.
{B.}Histological and immunohistochemical analyses
{C.}Cell culture and treatments :- Human NSCLC cell lines, NCI-H1975 (lung epithelial cells derived
from adenocarcinoma tissue; ATCC# CRL-5908) and NCI-H292 (a lymph node metastasis of
a pulmonary mucoepidermoid carcinoma; ATCC# CRL-1848), were obtained from the
American Type Culture Collection (ATCC, Manassas, VA, USA)..
{D.}siG6PD and transfection assay
{E.}Cell proliferation assay.
{F.}Clonogenic assay
{G}G6PD activity assay.
{H.}Scratch wound assay
{I.}Quantitative RT-PCR
{J}Statistical analysis
All the experiments were obtained from three independent experiments performed in triplicate and presented as the mean ± standard deviation (SD). The chi-square test (Ç2) and Student' t-test were used to examine the differences between categorical and quantitative variables. A two-sided difference
with a p-value less than 0.05 was considered statistically significant.
{A.}The investigators claim that the present study reported for the first time the high expression of G6PD in NSCLC {non-small cell lung cancer} tissues as compared to SCLC {small cell lung cancer} tissues.
{B.}G6PD{Glucose 6-phosphate dehydrogenase} expression was more intense in squamous
cell than adenocarcinoma. This finding was in line with the CU-DREAM analysis of NCBIís
GEO dataset program, targeting the over expression of G6PD protein in lung cancer tissues
compared to adjacent normal tissues.
{C.} Their study also demonstrated a positive correlation between the advanced stages of NSCLC
and G6PD over expression,in tune with previous reports of high levels of G6PD expression
in poor prognosis lung cancer patients
{D.} Findings from this investigation have implications for targeting the role of G6PD in carcinogenesis
and the development of novel strategies for lung cancer therapy.
{E.}The differences in G6PD levels in these two NSCLC cell lines{NCI-H292,NCI- H1975 cell}
positively correlated with their cell proliferation rates, suggesting the importance of G6PD
in the growth and advancement of NSCLC cells.
{F.}DHEA treatment and application of siG6PD reduced cell proliferation and colony formation in
NSCLS cells were in tune with the published literature.
{G.}The present study demonstrated that DHEA treatment and siG6PD application reduced NSCLC
cell migration in a scratch-wound assay which concurs with earlier published literature.
The authors is of the opinion that the underlying mechanisms might involve the suppression
of epithelial-mesenchymal transition(EMT) through E-cadherin activation.
{H} The authors have demonstrated that inhibition of G6PD activity and down-regulation of G6PD
mRNA expression elevated the Bax/Bcl-2 mRNA ratio, indicating an increasing susceptibility of
NSCLC cells to apoptosis.
{I} Hence, G6PD could be a potential therapeutic target for NCI-H292 cell lines.The authors stated
that the results indicate that suppression of G6PD by DHEA or siG6PD altered EMR through
the regulation of PPP in lung cancer cell lines.
Nil
Overall this is a well-written manuscript. However, I suggest authors consider comments below and in the next sections:
Please explain the rationale for this study well.
Cisplatin-based interventions are s a commonly used chemotherapy regimen in advanced patients. I suggest the authors look into how response/sensitivity towards Cisplatin is influenced by Inhibition of G6PD
I suggest authors also include additional markers of cellular redox homeostasis to make this study robust and further useful for the field.
Without proper controls, G6PD H-score calculations and resultant binning is not very helpful.
Sample selection and analysis:
Did these patients undergo any prior medical treatment? Evaluation and analysis of what types of treatments patients undergo before sample collection would be an important factor affecting metabolic programming. I suggest authors evaluate details of medical history, and rebin patients, and provide insights into how analyses were controlled.
I suggest authors also look at other enzymes upstream of G6PD to determine if this is a pathway-specific pattern.
Figure 2 (corresponding experiments) needs control samples.
Mention in figure legends: How many samples? How many repeats?
How do authors explain G6PD mRNA levels in NCI-H1975 and NCI-H292 Figure 4A?
Several more controls are needed to be included in the study to make findings robust.
All text and materials provided via this peer-review history page are made available under a Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.