Reviewer ’s report  
 
Title: Development of a transgenic mouse model of hepatocellular carcinoma with a liver  
fibrosis  background  
 
Version: 0 Date: 23 Jul 2015  
 
Reviewer: Eleonora Patsenker  
 
Reviewer's report:  
 
The present work describes the dev elopment of a new transgenic mouse model of HCC with a 
liver fibrosis background. Hydrodynamic transfection coupled with Sleeping Beauty transposon 
system expressing cMyc and short hairpin RNA downregulating p53 was used to prime the liver 
towards mutagene sis and in addition, chronic administration of CCL4 was applied in transgenic 
mice to induce chronic hepatic injury and fibrosis. As a result, hepatocarcinogenesis was 
significantly accelerated in fibrotic livers compared to non -fibrotic tumor controls. Th e 
conclusion is: the simplicity and the efficiency of the model has the potential to be used 
effectively in testing of HCC therapy and studies of hepatocarcinogenesis in fibrotic livers.  
The study is well designed and nicely performed, and the manuscript is clearly written. There is 
only one minor addition would be nice to see as a confirmation of technique performance: figure 
showing cMyc and p53 expression in the livers of control and tr ansgenic mice. The exact 
concentration of CCL4 used should be indicated in M&M (or it was 1ml/kg of 100% solution?). 
Also, the images for the publication could be of better quality.    
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, pl ease explain in your comments to the authors.  
 
Yes 
 
Are you able to assess any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require furth er 
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
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 s hares 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  
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 publication, my named report 
including any attachments I upload will be poste d on the website along with the authors' 
responses. I agree for my report to be made available under an Open Ac cess Creative Commons 
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments 
which I do not wish to be inclu ded 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: Development of a transgenic mouse model of hepatocellular carcinoma with a liver  
fibrosis  background  
 
Version: 0 Date: 23 Jul 2015  
 
Reviewer: Eleonora Patsenker  
 
Reviewer's report:  
 
The present work describes the dev elopment of a new transgenic mouse model of HCC with a 
liver fibrosis background. Hydrodynamic transfection coupled with Sleeping Beauty transposon 
system expressing cMyc and short hairpin RNA downregulating p53 was used to prime the liver 
towards mutagene sis and in addition, chronic administration of CCL4 was applied in transgenic 
mice to induce chronic hepatic injury and fibrosis. As a result, hepatocarcinogenesis was 
significantly accelerated in fibrotic livers compared to non -fibrotic tumor controls. Th e 
conclusion is: the simplicity and the efficiency of the model has the potential to be used 
effectively in testing of HCC therapy and studies of hepatocarcinogenesis in fibrotic livers.  
The study is well designed and nicely performed, and the manuscript is clearly written. There is 
only one minor addition would be nice to see as a confirmation of technique performance: figure 
showing cMyc and p53 expression in the livers of control and tr ansgenic mice. The exact 
concentration of CCL4 used should be indicated in M&M (or it was 1ml/kg of 100% solution?). 
Also, the images for the publication could be of better quality.    
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, pl ease explain in your comments to the authors.  
 
Yes 
 
Are you able to assess any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require furth er 
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
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 s hares 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  
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 publication, my named report 
including any attachments I upload will be poste d on the website along with the authors' 
responses. I agree for my report to be made available under an Open Ac cess Creative Commons 
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments 
which I do not wish to be inclu ded 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: Development of a transgenic mouse model of hepatocellular carcinoma with a liver  
fibrosis  background  
 
Version: 0 Date: 23 Jul 2015  
 
Reviewer: Eleonora Patsenker  
 
Reviewer's report:  
 
The present work describes the dev elopment of a new transgenic mouse model of HCC with a 
liver fibrosis background. Hydrodynamic transfection coupled with Sleeping Beauty transposon 
system expressing cMyc and short hairpin RNA downregulating p53 was used to prime the liver 
towards mutagene sis and in addition, chronic administration of CCL4 was applied in transgenic 
mice to induce chronic hepatic injury and fibrosis. As a result, hepatocarcinogenesis was 
significantly accelerated in fibrotic livers compared to non -fibrotic tumor controls. Th e 
conclusion is: the simplicity and the efficiency of the model has the potential to be used 
effectively in testing of HCC therapy and studies of hepatocarcinogenesis in fibrotic livers.  
The study is well designed and nicely performed, and the manuscript is clearly written. There is 
only one minor addition would be nice to see as a confirmation of technique performance: figure 
showing cMyc and p53 expression in the livers of control and tr ansgenic mice. The exact 
concentration of CCL4 used should be indicated in M&M (or it was 1ml/kg of 100% solution?). 
Also, the images for the publication could be of better quality.    
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, pl ease explain in your comments to the authors.  
 
Yes 
 
Are you able to assess any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require furth er 
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
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 s hares 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  
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 publication, my named report 
including any attachments I upload will be poste d on the website along with the authors' 
responses. I agree for my report to be made available under an Open Ac cess Creative Commons 
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments 
which I do not wish to be inclu ded 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