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: 
23 Nov 2020
Reviewer's report:
This study used immunohistochemistry to detect the expression of BRAF via
tissue microarrays in esophageal cancer samples, and determine its effect on
patient outcomes. They found BRAF overexpression was significantly correlated
with low Ki67 (P < 0.05) and shorter survival (P = 0.014). Cell culture study
showed BRAF knockdown reduce colony formation, cell migration and invasion.
Xenograft animal model showed BRAF inhibition suppressed tumor growth.
Major comments:
1. What is the histological type of esophageal cancer? For example, squamous
cell carcinoma or adenocarcinoma? The data should be presented in Table 1. 
2. Did the enrolled patients receive pre-operative chemotherapy or radiotherapy?
Or did they receive post-operative chemotherapy or radiotherapy? The data
should be presented in Table 1. In addition, the treatment modality should be
included in the survival analysis (both univariate and multivariate Cox
regression), because treatment modality is an important determinant of
prognosis. 
3. In Table 1 and Table 2, it is important to clarify the definition of AJCC
staging. Are they pathological staging (determined by surgical specimen) or
clinical staging (determined by imaging studies)?
4. The expression of Ki67 is strongly associated with tumor cell proliferation and
growth. However, this study showed a negative correlation between Ki67
expression and BRAF (Table 2). Note that weak BRAF expression correlated
with high Ki67. In other words, tumor with weak BRAF expression correlated
with increased tumor proliferation. This clinical data showed a conflict result
with that of the survival data (weak BRAF correlated with better survival) and
basic studies, including cell & animal studies (BRAF knock down correlated
with reduced tumor cell proliferation). The author should explain why there is
such a conflict. 
5. In the Cox regression analysis (Table 3), it is important to show whether the
variable is treated as continuous or categorical variable. In the categorical
variable, which group is treated as referent to calculate HR? 
6. In Fig. 2, please do subgroup Kaplan Meir survival analyses in AJCC stage II
and stage III patients, respectively, to show the association between BRAF
expression and survival of patients with each stage. 
Minor comments:
1. Esophageal cancer is not the most common malignant tumor. Please correct
the mistake in the "Abstract".
2. In result, "According to the seventh edition of the AJCC staging system, 5
cases were classified as phase I, 47 were phase II, 53 were phase III." The word
"phase" should be "stage".
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Publons’ privacy policy 
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No
Declaration of competing interests
Please complete a declaration of competing interests, considering the following
questions:
1
. 
Have you in the past five years received reimbursements, fees, funding, or
salary from an organisation that may in any way gain or lose financially
from the publication of this manuscript, either now or in the future?
2
. 
Do you hold any stocks or shares in an organisation that may in any way
gain or lose financially from the publication of this manuscript, either now
or in the future?
3
. 
Do you hold or are you currently applying for any patents relating to the
content of the manuscript?
4
. 
Have you received reimbursements, fees, funding, or salary from an
organization that holds or has applied for patents relating to the content of
the manuscript?
5
. 
Do you have any other financial competing interests?
6
. 
Do you have any non-financial competing interests in relation to this
paper?
If you can answer no to all of the above, write 'I declare that I have no competing
interests' below. If your reply is yes to any, please give details below.
I declare that I have no competing interests
Reviewer Publication Consent.
 I agree for my report to be made available
under an Open Access Creative Commons CC-BY License (
http://creativecommons.org/licenses/by/4.0
) if this manuscript is accepted for
publication. Any comments that I do not wish to be included in the published
report have been included as confidential comments to the editor, which will not
be published.
If you are not happy for us to publish this report, please contact the editorial
office before completing the review.
If you wish, you can include your name in your published report. Please note
you must decide whether to include your name at the start of the process and
confirm this decision whenever prompted. If you change your decision later, we
will not be able to publish your name.
I agree to the terms of the CC-BY 4.0 license; please 
do not
 publish my name
with my report. (default)
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
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: 
23 Nov 2020
Reviewer's report:
This study used immunohistochemistry to detect the expression of BRAF via
tissue microarrays in esophageal cancer samples, and determine its effect on
patient outcomes. They found BRAF overexpression was significantly correlated
with low Ki67 (P < 0.05) and shorter survival (P = 0.014). Cell culture study
showed BRAF knockdown reduce colony formation, cell migration and invasion.
Xenograft animal model showed BRAF inhibition suppressed tumor growth.
Major comments:
1. What is the histological type of esophageal cancer? For example, squamous
cell carcinoma or adenocarcinoma? The data should be presented in Table 1. 
2. Did the enrolled patients receive pre-operative chemotherapy or radiotherapy?
Or did they receive post-operative chemotherapy or radiotherapy? The data
should be presented in Table 1. In addition, the treatment modality should be
included in the survival analysis (both univariate and multivariate Cox
regression), because treatment modality is an important determinant of
prognosis. 
3. In Table 1 and Table 2, it is important to clarify the definition of AJCC
staging. Are they pathological staging (determined by surgical specimen) or
clinical staging (determined by imaging studies)?
4. The expression of Ki67 is strongly associated with tumor cell proliferation and
growth. However, this study showed a negative correlation between Ki67
expression and BRAF (Table 2). Note that weak BRAF expression correlated
with high Ki67. In other words, tumor with weak BRAF expression correlated
with increased tumor proliferation. This clinical data showed a conflict result
with that of the survival data (weak BRAF correlated with better survival) and
basic studies, including cell & animal studies (BRAF knock down correlated
with reduced tumor cell proliferation). The author should explain why there is
such a conflict. 
5. In the Cox regression analysis (Table 3), it is important to show whether the
variable is treated as continuous or categorical variable. In the categorical
variable, which group is treated as referent to calculate HR? 
6. In Fig. 2, please do subgroup Kaplan Meir survival analyses in AJCC stage II
and stage III patients, respectively, to show the association between BRAF
expression and survival of patients with each stage. 
Minor comments:
1. Esophageal cancer is not the most common malignant tumor. Please correct
the mistake in the "Abstract".
2. In result, "According to the seventh edition of the AJCC staging system, 5
cases were classified as phase I, 47 were phase II, 53 were phase III." The word
"phase" should be "stage".
Publons Reviewer Recognition
. Springer Nature can send verification of this
review directly to Publons (a subsidiary of Clarivate Analytics). If you would
like to take advantage of this service, please click on the “Yes” option below.
Your name, email address, title of the reviewed manuscript, name of the journal,
and date of your review submission (the “Review Data”) will then be transmitted
to Publons upon publication of the manuscript. If you have already registered at
Publons, they will notify you of the receipt of this review and update your profile
as per your settings and their policy. If you are not registered with Publons, you
will receive an email from them asking you to register in order for them to be
able to recognize your review on your new profile page. Publons may use the
Review Data to generate derivative metadata for the benefit of Publons and you
as a reviewer, carefully considering the sensitivity of such information. For
example, Publons may verify your record as a reviewer by updating your profile
published on its webservice if you have registered for such service or help
editors to identify candidate reviewers. Please find the details of processing in
Publons’ privacy policy 
https://publons.com/about/terms
No
Declaration of competing interests
Please complete a declaration of competing interests, considering the following
questions:
1
. 
Have you in the past five years received reimbursements, fees, funding, or
salary from an organisation that may in any way gain or lose financially
from the publication of this manuscript, either now or in the future?
2
. 
Do you hold any stocks or shares in an organisation that may in any way
gain or lose financially from the publication of this manuscript, either now
or in the future?
3
. 
Do you hold or are you currently applying for any patents relating to the
content of the manuscript?
4
. 
Have you received reimbursements, fees, funding, or salary from an
organization that holds or has applied for patents relating to the content of
the manuscript?
5
. 
Do you have any other financial competing interests?
6
. 
Do you have any non-financial competing interests in relation to this
paper?
If you can answer no to all of the above, write 'I declare that I have no competing
interests' below. If your reply is yes to any, please give details below.
I declare that I have no competing interests
Reviewer Publication Consent.
 I agree for my report to be made available
under an Open Access Creative Commons CC-BY License (
http://creativecommons.org/licenses/by/4.0
) if this manuscript is accepted for
publication. Any comments that I do not wish to be included in the published
report have been included as confidential comments to the editor, which will not
be published.
If you are not happy for us to publish this report, please contact the editorial
office before completing the review.
If you wish, you can include your name in your published report. Please note
you must decide whether to include your name at the start of the process and
confirm this decision whenever prompted. If you change your decision later, we
will not be able to publish your name.
I agree to the terms of the CC-BY 4.0 license; please 
do not
 publish my name
with my report. (default)
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
Please review this paper.
user
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: 
23 Nov 2020
Reviewer's report:
This study used immunohistochemistry to detect the expression of BRAF via
tissue microarrays in esophageal cancer samples, and determine its effect on
patient outcomes. They found BRAF overexpression was significantly correlated
with low Ki67 (P < 0.05) and shorter survival (P = 0.014). Cell culture study
showed BRAF knockdown reduce colony formation, cell migration and invasion.
Xenograft animal model showed BRAF inhibition suppressed tumor growth.
Major comments:
1. What is the histological type of esophageal cancer? For example, squamous
cell carcinoma or adenocarcinoma? The data should be presented in Table 1. 
2. Did the enrolled patients receive pre-operative chemotherapy or radiotherapy?
Or did they receive post-operative chemotherapy or radiotherapy? The data
should be presented in Table 1. In addition, the treatment modality should be
included in the survival analysis (both univariate and multivariate Cox
regression), because treatment modality is an important determinant of
prognosis. 
3. In Table 1 and Table 2, it is important to clarify the definition of AJCC
staging. Are they pathological staging (determined by surgical specimen) or
clinical staging (determined by imaging studies)?
4. The expression of Ki67 is strongly associated with tumor cell proliferation and
growth. However, this study showed a negative correlation between Ki67
expression and BRAF (Table 2). Note that weak BRAF expression correlated
with high Ki67. In other words, tumor with weak BRAF expression correlated
with increased tumor proliferation. This clinical data showed a conflict result
with that of the survival data (weak BRAF correlated with better survival) and
basic studies, including cell & animal studies (BRAF knock down correlated
with reduced tumor cell proliferation). The author should explain why there is
such a conflict. 
5. In the Cox regression analysis (Table 3), it is important to show whether the
variable is treated as continuous or categorical variable. In the categorical
variable, which group is treated as referent to calculate HR? 
6. In Fig. 2, please do subgroup Kaplan Meir survival analyses in AJCC stage II
and stage III patients, respectively, to show the association between BRAF
expression and survival of patients with each stage. 
Minor comments:
1. Esophageal cancer is not the most common malignant tumor. Please correct
the mistake in the "Abstract".
2. In result, "According to the seventh edition of the AJCC staging system, 5
cases were classified as phase I, 47 were phase II, 53 were phase III." The word
"phase" should be "stage".
Publons Reviewer Recognition
. Springer Nature can send verification of this
review directly to Publons (a subsidiary of Clarivate Analytics). If you would
like to take advantage of this service, please click on the “Yes” option below.
Your name, email address, title of the reviewed manuscript, name of the journal,
and date of your review submission (the “Review Data”) will then be transmitted
to Publons upon publication of the manuscript. If you have already registered at
Publons, they will notify you of the receipt of this review and update your profile
as per your settings and their policy. If you are not registered with Publons, you
will receive an email from them asking you to register in order for them to be
able to recognize your review on your new profile page. Publons may use the
Review Data to generate derivative metadata for the benefit of Publons and you
as a reviewer, carefully considering the sensitivity of such information. For
example, Publons may verify your record as a reviewer by updating your profile
published on its webservice if you have registered for such service or help
editors to identify candidate reviewers. Please find the details of processing in
Publons’ privacy policy 
https://publons.com/about/terms
No
Declaration of competing interests
Please complete a declaration of competing interests, considering the following
questions:
1
. 
Have you in the past five years received reimbursements, fees, funding, or
salary from an organisation that may in any way gain or lose financially
from the publication of this manuscript, either now or in the future?
2
. 
Do you hold any stocks or shares in an organisation that may in any way
gain or lose financially from the publication of this manuscript, either now
or in the future?
3
. 
Do you hold or are you currently applying for any patents relating to the
content of the manuscript?
4
. 
Have you received reimbursements, fees, funding, or salary from an
organization that holds or has applied for patents relating to the content of
the manuscript?
5
. 
Do you have any other financial competing interests?
6
. 
Do you have any non-financial competing interests in relation to this
paper?
If you can answer no to all of the above, write 'I declare that I have no competing
interests' below. If your reply is yes to any, please give details below.
I declare that I have no competing interests
Reviewer Publication Consent.
 I agree for my report to be made available
under an Open Access Creative Commons CC-BY License (
http://creativecommons.org/licenses/by/4.0
) if this manuscript is accepted for
publication. Any comments that I do not wish to be included in the published
report have been included as confidential comments to the editor, which will not
be published.
If you are not happy for us to publish this report, please contact the editorial
office before completing the review.
If you wish, you can include your name in your published report. Please note
you must decide whether to include your name at the start of the process and
confirm this decision whenever prompted. If you change your decision later, we
will not be able to publish your name.
I agree to the terms of the CC-BY 4.0 license; please 
do not
 publish my name
with my report. (default)
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
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