Review for 10.1186/s12876-021-01671-2 (2021)

Reviewer’s report
Title: 
V-Raf murine sarcoma viral oncogene homolog B1 (BRAF) as a
prognostic biomarker of poor outcomes in esophageal cancer patients
Version: 
0 
Date: 
06 Jan 2021
Reviewer's report:
The current study investigated the prognostic role of BRAF expression in
esophageal cancer using TMA consisted of 105 cancer tissues, and tested several
cellular works as well as xenograft mouse model with TE-1 cell.
Even though the result looks simple and straightforward, any correlation
between BRAF mutation / EGFR pathway (proposed as hypothesis) and over-
expression of BRAF (as main analysis) was not presented in this study. If there
is any significant correlation, the results looks consistent to previous hypothesis
but may not be novel findings. Otherwise, authors should explain possible
mechanism of over-expression regardless of mutation. Besides, other several
issues below be addressed.
#1. please clarify the basic histology of Esophageal cancer in this study: 
were
they all squamous cell carcinoma or adenoca?
#2. please clarify the definition of "strongly" vs. "weakly" expression of BRAF,
probably based on authors' interpretation of TMA (0,1,2, and 3). Were all
samples dichotomized 1 (weakly positive) or 3 (strongly positive) without 0
(negative) and 2 (positive)?
#3. What does "adjacent-to-carcinoma" represent? is it paired normal or possible
precancerous lesion? please describe that definition or how to retrieve that
sample clearly in methods section.
#4. What about the expression profile of "adjacent-to-carcinoma" tissue? 
This
may be lower expression rate that tumor because it should be less aggressive or
normal compared with tumor itself. Since they were coupled with corresponding
tumor, the paired comparative analysis should be valuable. 
#5. There is no methodology of siRNA work. e.g. vendor? what is the primer?
#6. There was no WB work for BRAF expression status in cell line after siRNA
treatment. please present KO status of BRAF before and after siRNA. WB of not
only BRAF but also Ki-67/P53 could be very useful. 
#7. BRAF expression status and any pathologic response data of retrieved mass
from xenograft model is required to evaluate BRAF inhibitor. please describe the
detailed information of BRAF inhibitor in methods as well.
#8. In addition to OS, disease free or progression free interval could be more
helpful
#9. Figure 3A needs p value. Were there any significant change of volume at a
certain time point?
#10. The sample size looks not enough to validate the prognostic difference. Any
type of external validation using TCGA or other public dataset could be helpful,
even tumor vs. normal using mutation, RNA or RPPA expression.
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