Reviewer ’s report  
 
Title: Upregulation of musashi1 increases malignancy of hepatocellular carcinoma via the 
Wnt/β-catenin signaling pathway and predicts a poor prognosis  
 
Version: 0 Date: 18 Jul 2019  
 
Reviewer: Wadie Fernandez  
 
Reviewer's report:  
 
The manuscript titled "Musashi1 predict poor diagnosis and malignant transformation in 
hepatocellular carcinoma by activation of  Wnt/B -catenin signaling pathway" by Liu et. al. 
describes Musashi -1 as a prognostic factor for HCC that affects cancer cell viability, migration, 
and invasion via activation of the Wnt/B -catenin pathway; and, Musashi -1 also induces CD44 
which is a stem -cell marker. Musashi -1 is suggested to be a potential therapeutic target against 
HCC.  
 
 
Major comments:  
 
1. Figure 1A: case 3 para -cancer sample is extremely positive for Musashi -1. Is this image 
erroneous?  
 
2. Please elaborate in your results section as to why  you are testing the WNT -B/catenin 
pathway in relation to Musashi -1. What's the basis to test this? There is no background 
explain this.  
 
3. Another shRNA against Musashi -1 should be used to rule out off -target effects. In this 
manuscript only one shRNA is used.  
 
4. Please add western blots showing CD44 levels in cells expressing shControl and 
shMusashi -1; and in cells overexpressing Musashi -1 to complement your other CD44 
data.  
 
5. Please add survival plots of patients divided in the following groups: Musashi -
high/CD44 -high and musashi -low/CD44 -low. 
 
6. In the discussion section, please add a possible mechanism(s) as to how Musashi -1 
induces WNt/B -catenin and how Musashi -1 induces CD44.  
 
7. Figure legends need to be more descriptive. There is no real information  in any of them.  
 
 
Minor comments:  
 
1. Need to add a reference in text lines 1 -2. 
 
2. Please correct all grammatical errors  
 
3. Please revise the fonts in all figures to make then readable.  
 
4. DO not use the word "obviously" since nothing that is tested can be  obvious.  
 
5. Add scale bars to all images in figures.  
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comme nts 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 adequate ly supported by the data shown?  
If not, please explain in your c omments to the authors.  
 
Yes 
 
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.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Needs some language corrections before being published  
 
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, f ees, funding, or salary 
from an organisation that may in any way gain or lose financially from the 
public ation 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 curre ntly 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 g ive details below.  
 
I declare that I have no competing interests  
 
I agree to the open peer review policy of the journal. I understan d that my name will be included 
on my report to the authors and, if the manuscript is accepted for publication, my named repor t 
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 in cluded 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: Upregulation of musashi1 increases malignancy of hepatocellular carcinoma via the 
Wnt/β-catenin signaling pathway and predicts a poor prognosis  
 
Version: 0 Date: 18 Jul 2019  
 
Reviewer: Wadie Fernandez  
 
Reviewer's report:  
 
The manuscript titled "Musashi1 predict poor diagnosis and malignant transformation in 
hepatocellular carcinoma by activation of  Wnt/B -catenin signaling pathway" by Liu et. al. 
describes Musashi -1 as a prognostic factor for HCC that affects cancer cell viability, migration, 
and invasion via activation of the Wnt/B -catenin pathway; and, Musashi -1 also induces CD44 
which is a stem -cell marker. Musashi -1 is suggested to be a potential therapeutic target against 
HCC.  
 
 
Major comments:  
 
1. Figure 1A: case 3 para -cancer sample is extremely positive for Musashi -1. Is this image 
erroneous?  
 
2. Please elaborate in your results section as to why  you are testing the WNT -B/catenin 
pathway in relation to Musashi -1. What's the basis to test this? There is no background 
explain this.  
 
3. Another shRNA against Musashi -1 should be used to rule out off -target effects. In this 
manuscript only one shRNA is used.  
 
4. Please add western blots showing CD44 levels in cells expressing shControl and 
shMusashi -1; and in cells overexpressing Musashi -1 to complement your other CD44 
data.  
 
5. Please add survival plots of patients divided in the following groups: Musashi -
high/CD44 -high and musashi -low/CD44 -low. 
 
6. In the discussion section, please add a possible mechanism(s) as to how Musashi -1 
induces WNt/B -catenin and how Musashi -1 induces CD44.  
 
7. Figure legends need to be more descriptive. There is no real information  in any of them.  
 
 
Minor comments:  
 
1. Need to add a reference in text lines 1 -2. 
 
2. Please correct all grammatical errors  
 
3. Please revise the fonts in all figures to make then readable.  
 
4. DO not use the word "obviously" since nothing that is tested can be  obvious.  
 
5. Add scale bars to all images in figures.  
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comme nts 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 adequate ly supported by the data shown?  
If not, please explain in your c omments to the authors.  
 
Yes 
 
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.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Needs some language corrections before being published  
 
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, f ees, funding, or salary 
from an organisation that may in any way gain or lose financially from the 
public ation 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 curre ntly 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 g ive details below.  
 
I declare that I have no competing interests  
 
I agree to the open peer review policy of the journal. I understan d that my name will be included 
on my report to the authors and, if the manuscript is accepted for publication, my named repor t 
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 in cluded 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: Upregulation of musashi1 increases malignancy of hepatocellular carcinoma via the 
Wnt/β-catenin signaling pathway and predicts a poor prognosis  
 
Version: 0 Date: 18 Jul 2019  
 
Reviewer: Wadie Fernandez  
 
Reviewer's report:  
 
The manuscript titled "Musashi1 predict poor diagnosis and malignant transformation in 
hepatocellular carcinoma by activation of  Wnt/B -catenin signaling pathway" by Liu et. al. 
describes Musashi -1 as a prognostic factor for HCC that affects cancer cell viability, migration, 
and invasion via activation of the Wnt/B -catenin pathway; and, Musashi -1 also induces CD44 
which is a stem -cell marker. Musashi -1 is suggested to be a potential therapeutic target against 
HCC.  
 
 
Major comments:  
 
1. Figure 1A: case 3 para -cancer sample is extremely positive for Musashi -1. Is this image 
erroneous?  
 
2. Please elaborate in your results section as to why  you are testing the WNT -B/catenin 
pathway in relation to Musashi -1. What's the basis to test this? There is no background 
explain this.  
 
3. Another shRNA against Musashi -1 should be used to rule out off -target effects. In this 
manuscript only one shRNA is used.  
 
4. Please add western blots showing CD44 levels in cells expressing shControl and 
shMusashi -1; and in cells overexpressing Musashi -1 to complement your other CD44 
data.  
 
5. Please add survival plots of patients divided in the following groups: Musashi -
high/CD44 -high and musashi -low/CD44 -low. 
 
6. In the discussion section, please add a possible mechanism(s) as to how Musashi -1 
induces WNt/B -catenin and how Musashi -1 induces CD44.  
 
7. Figure legends need to be more descriptive. There is no real information  in any of them.  
 
 
Minor comments:  
 
1. Need to add a reference in text lines 1 -2. 
 
2. Please correct all grammatical errors  
 
3. Please revise the fonts in all figures to make then readable.  
 
4. DO not use the word "obviously" since nothing that is tested can be  obvious.  
 
5. Add scale bars to all images in figures.  
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comme nts 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 adequate ly supported by the data shown?  
If not, please explain in your c omments to the authors.  
 
Yes 
 
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.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Needs some language corrections before being published  
 
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, f ees, funding, or salary 
from an organisation that may in any way gain or lose financially from the 
public ation 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 curre ntly 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 g ive details below.  
 
I declare that I have no competing interests  
 
I agree to the open peer review policy of the journal. I understan d that my name will be included 
on my report to the authors and, if the manuscript is accepted for publication, my named repor t 
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 in cluded as confidential comments 
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
assistant