Review for 10.1186/s12876-020-01556-w (2020)

Reviewer’s report
Title: 
Novel stromal biomarker screening in pancreatic cancer patients using the
in vitro cancer-stromal interaction model
Version: 
0 
Date: 
06 Sep 2020
Reviewer's report:
The authors of the manuscript "Novel stromal biomarker screening in pancreatic
cancer patients using the in vitro cancer-stromal interaction mode" have explored
an interesting domain of the pancreatic stromal biomarker for prediction of
pancreatic adenocarcinoma and found DIAPH3 to be an important prognostic
marker.
I have the following comments to make:
1. This line "High gene expression was based on > median mRNA expression
value
for each gene" is not explainable and needs to be clarified
2. The authors on survival analysis found 6 genes as significant predictors for
survival/poor prognosis. Yet, while analyzing DIAPH3 expression was found in
the cancerous tissue only, although the survival analysis was also done in 135
PDAC patients only. This is confusing and needs to be clarified. 
3. How were low and high levels of the various genes determined is not
mentioned.
4. While, strong expression of DIAPH3 was associated with poor overall
survival, survival depends on a host of factors including tumor size, lymph node
metastasis etc. 
Were these confounding factors accounted for during the survival
curve analysis?
5. Moreover, strong expression of DIAPH3 group had more lymph node
metastasis. How can the authors predict that this increased lymph node
metastasis is because of increased expression of DIAPH3 and not just a
confounding factor?
6. The conclusion of the abstract does not match with the study outcome and
needs to be modified. 
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Is the study design appropriate to answer the research question (including the use
of appropriate controls), and are the conclusions supported by the evidence
presented?
Yes
Are the methods sufficiently described to allow the study to be repeated?
Yes
Is the use of statistics and treatment of uncertainties appropriate?
No
Is the presentation of the work clear?
No
Are the images in this manuscript (including electrophoretic gels and blots) free
from apparent manipulation?
Yes