Reviewer ’s report  
Title: Apatinib affect VEGF -mediated cell proliferation, m igration, invasion via blocking 
VEGFR2/MEK/ERK in cho langiocarcinoma cell of QBC939  
 
Version: 0 Date: 31 Jan 2018  
 
Reviewer: Giuliana Cavalloni  
 
Reviewer's report:  
In this paper the authors demonstrated that the tyrosine kinase inhibitor that selectively the 
VEGFR2 inhibits proliferation, invasion and migration in one in vitro biliary tumor model.  
 
Major comments:  
1) The work is descriptive and adds, compared to previous work by Peng H et al., (Oncotarget 
2016), only data relating to the inhibition of migration and invasion after apatinib treatment. 
The mechanis m of action is not further investigated; for example which molecules are 
involved (i.e. src signaling pathway also involved in adhesion processes, MMPs in invasion 
processes…..)  
 
2) The data were obtained using only one in vitro model of biliary tumors (in tra or extraepatic 
cholangiocarcinoma?). In my opinion, only one "aspecific" model (it is not specified from 
which istotype the QBC939 cells derived), in not exhaustive; the authors should extend the 
study to other commercially available cell lines, that s hould be of the same istotype of the 
cells used in this work in order to confirm your data and to deepen, at least the invasion, 
migration and adhesion mechanisms.  
 
3) The data add very few further information compared to another preclinical study (Peng S et 
al., Cancer Letter 2016) in in vitro and in vivo biliary tumor model. Since that VEGFR2 is 
xpressed also in endothelial cells, It could be interesting investigated the role of apatinib in 
tumor angiogenesis in vitro by using HUVEC cells and in vivo anal yzing tumor murine 
angiogenesis in in vivo model.  
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
Yes 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
Yes 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
No 
Are you able to assess any statistics in the manus cript or would you recommend an 
additional statistical review?  
If an additional statistical review is recommended, please speci fy what aspects require further 
assessment in your comments to the editors.  
Not relevant to this manuscript  
Quality of written English  
Please indicate the quality of language in the manuscript:  
Needs some language corrections before being published  
Decla ration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
1. Have you in the past five years received reimbursements, f ees, funding, or salary from an 
organisation that may in any way gain or l ose financially from the publication of this  
manuscript, either now or in the future?  
2. Do you hold any stocks or shares in an organisation t hat 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 y es to any, please give details below.  
 
I declare that I have no competing interests  
 
I agree to the open peer review policy of the journal. I understand that my name will be included 
on my report to the authors and, if the manuscript is accepted for publicati on, my named report 
including any attachments I upload will be posted on the website along with the authors' 
responses. 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/). I understand that any comments 
which I do not wish to be included in my named report can be included as confidential comments 
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
Reviewer ’s report  
Title: Apatinib affect VEGF -mediated cell proliferation, m igration, invasion via blocking 
VEGFR2/MEK/ERK in cho langiocarcinoma cell of QBC939  
 
Version: 0 Date: 31 Jan 2018  
 
Reviewer: Giuliana Cavalloni  
 
Reviewer's report:  
In this paper the authors demonstrated that the tyrosine kinase inhibitor that selectively the 
VEGFR2 inhibits proliferation, invasion and migration in one in vitro biliary tumor model.  
 
Major comments:  
1) The work is descriptive and adds, compared to previous work by Peng H et al., (Oncotarget 
2016), only data relating to the inhibition of migration and invasion after apatinib treatment. 
The mechanis m of action is not further investigated; for example which molecules are 
involved (i.e. src signaling pathway also involved in adhesion processes, MMPs in invasion 
processes…..)  
 
2) The data were obtained using only one in vitro model of biliary tumors (in tra or extraepatic 
cholangiocarcinoma?). In my opinion, only one "aspecific" model (it is not specified from 
which istotype the QBC939 cells derived), in not exhaustive; the authors should extend the 
study to other commercially available cell lines, that s hould be of the same istotype of the 
cells used in this work in order to confirm your data and to deepen, at least the invasion, 
migration and adhesion mechanisms.  
 
3) The data add very few further information compared to another preclinical study (Peng S et 
al., Cancer Letter 2016) in in vitro and in vivo biliary tumor model. Since that VEGFR2 is 
xpressed also in endothelial cells, It could be interesting investigated the role of apatinib in 
tumor angiogenesis in vitro by using HUVEC cells and in vivo anal yzing tumor murine 
angiogenesis in in vivo model.  
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
Yes 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
Yes 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
No 
Are you able to assess any statistics in the manus cript or would you recommend an 
additional statistical review?  
If an additional statistical review is recommended, please speci fy what aspects require further 
assessment in your comments to the editors.  
Not relevant to this manuscript  
Quality of written English  
Please indicate the quality of language in the manuscript:  
Needs some language corrections before being published  
Decla ration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
1. Have you in the past five years received reimbursements, f ees, funding, or salary from an 
organisation that may in any way gain or l ose financially from the publication of this  
manuscript, either now or in the future?  
2. Do you hold any stocks or shares in an organisation t hat 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 y es to any, please give details below.  
 
I declare that I have no competing interests  
 
I agree to the open peer review policy of the journal. I understand that my name will be included 
on my report to the authors and, if the manuscript is accepted for publicati on, my named report 
including any attachments I upload will be posted on the website along with the authors' 
responses. 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/). I understand that any comments 
which I do not wish to be included in my named report can be included as confidential comments 
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
Please review this paper.
user
Reviewer ’s report  
Title: Apatinib affect VEGF -mediated cell proliferation, m igration, invasion via blocking 
VEGFR2/MEK/ERK in cho langiocarcinoma cell of QBC939  
 
Version: 0 Date: 31 Jan 2018  
 
Reviewer: Giuliana Cavalloni  
 
Reviewer's report:  
In this paper the authors demonstrated that the tyrosine kinase inhibitor that selectively the 
VEGFR2 inhibits proliferation, invasion and migration in one in vitro biliary tumor model.  
 
Major comments:  
1) The work is descriptive and adds, compared to previous work by Peng H et al., (Oncotarget 
2016), only data relating to the inhibition of migration and invasion after apatinib treatment. 
The mechanis m of action is not further investigated; for example which molecules are 
involved (i.e. src signaling pathway also involved in adhesion processes, MMPs in invasion 
processes…..)  
 
2) The data were obtained using only one in vitro model of biliary tumors (in tra or extraepatic 
cholangiocarcinoma?). In my opinion, only one "aspecific" model (it is not specified from 
which istotype the QBC939 cells derived), in not exhaustive; the authors should extend the 
study to other commercially available cell lines, that s hould be of the same istotype of the 
cells used in this work in order to confirm your data and to deepen, at least the invasion, 
migration and adhesion mechanisms.  
 
3) The data add very few further information compared to another preclinical study (Peng S et 
al., Cancer Letter 2016) in in vitro and in vivo biliary tumor model. Since that VEGFR2 is 
xpressed also in endothelial cells, It could be interesting investigated the role of apatinib in 
tumor angiogenesis in vitro by using HUVEC cells and in vivo anal yzing tumor murine 
angiogenesis in in vivo model.  
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
Yes 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
Yes 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
No 
Are you able to assess any statistics in the manus cript or would you recommend an 
additional statistical review?  
If an additional statistical review is recommended, please speci fy what aspects require further 
assessment in your comments to the editors.  
Not relevant to this manuscript  
Quality of written English  
Please indicate the quality of language in the manuscript:  
Needs some language corrections before being published  
Decla ration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
1. Have you in the past five years received reimbursements, f ees, funding, or salary from an 
organisation that may in any way gain or l ose financially from the publication of this  
manuscript, either now or in the future?  
2. Do you hold any stocks or shares in an organisation t hat 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 y es to any, please give details below.  
 
I declare that I have no competing interests  
 
I agree to the open peer review policy of the journal. I understand that my name will be included 
on my report to the authors and, if the manuscript is accepted for publicati on, my named report 
including any attachments I upload will be posted on the website along with the authors' 
responses. 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/). I understand that any comments 
which I do not wish to be included in my named report can be included as confidential comments 
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
assistant