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: 04 Aug 2019  
 
Reviewer: Kais Zakharia  
 
Reviewer's report:  
 
Please include all comments for the authors in this box rather than uploadi ng your report as an 
attachment. Please only upload as attachments annotated versions of manuscripts, graphs, 
supporting materials or other aspects of your report which cannot be included in a text format.  
 
Please overwrite this text when adding your commen ts to the authors.  
 
Musashi1 predict poor prognosis and malignant transformation in hepatocellular carcinoma by 
activation of Wnt/β -catenin signaling pathway  
 
The authors looked at the Musashi1, which has noted to contribute to cancer progression in 
severa l cancers, in HCC.  
 
 
In the introduction  
 
Authors first showed that musashi1 was overexpressed in HCC tumors/cell lines compared to 
normal adjacent tissue. Figure 1A shows this finding. I suggest that authors quantify these 
findings and show them in a figu re in addition to what they showed (something similar to Fig 1 
C and D). In figure 1D, expression of musashi1 was increased in HCC cell lines compared to 
normal liver cells. P value should be reported.  
 
Later authors showed that musashi knockdown is associated with decreased cell viability and 
musashi overexpression is associated with increased viability. Two assays were used for this 
purpose which is good however we usually prefer to see this effect in m ore than two cells 
(usually three cell lines minimum specially that authors have four cell lines that showed 
overexpression of musashi1).  
 
The authors showed that the effect of musashi1 in HCC is through Wnt/β -catenin signaling 
pathway. This has been repo rted before. Chen K et. al. Liver Int. 2015 Mar;35(3):986 -98. doi: 
10.1111/liv.12458. Epub 2014 Jan 29.  
 
The authors showed that musashi1 overexpression is associated with poor prognosis of HCC 
patients. This is an interesting finding. Lu He et. al. showed  that musashi2, but not musashi1, 
was associated with poor prognosis. I think it worth discussing this contradictory finding in the 
current study. (J Cell Mol Med. 2014 Jan; 18(1): 49 -58). 
The manuscript is poorly written with numerous grammar mistakes (s ome sentences are not even 
understandable).  I will mention only few examples below.   
 
 
Examples:  
 
Page 2, line 51 -52: Authors mean to say PLC has become third cause of cancer related death 
worldwide however two mistakes are noted in this sentence: 1) the wo rd "which" is referring to 
morbidity and mortality in the way they stated it which is not what they mean. 2) They should 
add something like "cause/reason/etiology of" in between "third" and cancer -related death" 
otherwise the sentence doesn't make sense.  
 
Also in line 52: if primary liver cancer to be abbreviated as PLC, this abbreviation should be 
used later.   
 
Page 3, line 4 -5: authors started introducing Musashi1 gene and then they started describing the 
protein and stated "It has been identified as an e volutionarily conserved group of RNA binding 
protein and used as a molecular marker of neural stem cells."  
 
"It" is referring to the gene but they are describing the protein! This should be corrected.  
 
Page 3, line 7: "musashi" is spelled without "1"  
 
Page 6 , line 10: "were chose" should read were chosen.  
 
Throughout the manuscript: I got a sense that the authors naming the gene musashi1 and the 
protein musashi -1. If this is correct, those two words were used interchangeably throughout the 
manuscript i.e. sho uld be consistent.  
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the w ork 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.  
 
Yes 
 
Are yo u 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:  
 
Not suitable for publication unless extensively edited  
 
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 
publi cation 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 curr ently 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 understan d that my name will be included 
on my report to the authors and, if the manuscript is accepted for publication, my named repo rt 
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 understan d 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: 04 Aug 2019  
 
Reviewer: Kais Zakharia  
 
Reviewer's report:  
 
Please include all comments for the authors in this box rather than uploadi ng your report as an 
attachment. Please only upload as attachments annotated versions of manuscripts, graphs, 
supporting materials or other aspects of your report which cannot be included in a text format.  
 
Please overwrite this text when adding your commen ts to the authors.  
 
Musashi1 predict poor prognosis and malignant transformation in hepatocellular carcinoma by 
activation of Wnt/β -catenin signaling pathway  
 
The authors looked at the Musashi1, which has noted to contribute to cancer progression in 
severa l cancers, in HCC.  
 
 
In the introduction  
 
Authors first showed that musashi1 was overexpressed in HCC tumors/cell lines compared to 
normal adjacent tissue. Figure 1A shows this finding. I suggest that authors quantify these 
findings and show them in a figu re in addition to what they showed (something similar to Fig 1 
C and D). In figure 1D, expression of musashi1 was increased in HCC cell lines compared to 
normal liver cells. P value should be reported.  
 
Later authors showed that musashi knockdown is associated with decreased cell viability and 
musashi overexpression is associated with increased viability. Two assays were used for this 
purpose which is good however we usually prefer to see this effect in m ore than two cells 
(usually three cell lines minimum specially that authors have four cell lines that showed 
overexpression of musashi1).  
 
The authors showed that the effect of musashi1 in HCC is through Wnt/β -catenin signaling 
pathway. This has been repo rted before. Chen K et. al. Liver Int. 2015 Mar;35(3):986 -98. doi: 
10.1111/liv.12458. Epub 2014 Jan 29.  
 
The authors showed that musashi1 overexpression is associated with poor prognosis of HCC 
patients. This is an interesting finding. Lu He et. al. showed  that musashi2, but not musashi1, 
was associated with poor prognosis. I think it worth discussing this contradictory finding in the 
current study. (J Cell Mol Med. 2014 Jan; 18(1): 49 -58). 
The manuscript is poorly written with numerous grammar mistakes (s ome sentences are not even 
understandable).  I will mention only few examples below.   
 
 
Examples:  
 
Page 2, line 51 -52: Authors mean to say PLC has become third cause of cancer related death 
worldwide however two mistakes are noted in this sentence: 1) the wo rd "which" is referring to 
morbidity and mortality in the way they stated it which is not what they mean. 2) They should 
add something like "cause/reason/etiology of" in between "third" and cancer -related death" 
otherwise the sentence doesn't make sense.  
 
Also in line 52: if primary liver cancer to be abbreviated as PLC, this abbreviation should be 
used later.   
 
Page 3, line 4 -5: authors started introducing Musashi1 gene and then they started describing the 
protein and stated "It has been identified as an e volutionarily conserved group of RNA binding 
protein and used as a molecular marker of neural stem cells."  
 
"It" is referring to the gene but they are describing the protein! This should be corrected.  
 
Page 3, line 7: "musashi" is spelled without "1"  
 
Page 6 , line 10: "were chose" should read were chosen.  
 
Throughout the manuscript: I got a sense that the authors naming the gene musashi1 and the 
protein musashi -1. If this is correct, those two words were used interchangeably throughout the 
manuscript i.e. sho uld be consistent.  
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the w ork 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.  
 
Yes 
 
Are yo u 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:  
 
Not suitable for publication unless extensively edited  
 
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 
publi cation 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 curr ently 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 understan d that my name will be included 
on my report to the authors and, if the manuscript is accepted for publication, my named repo rt 
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 understan d 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: 04 Aug 2019  
 
Reviewer: Kais Zakharia  
 
Reviewer's report:  
 
Please include all comments for the authors in this box rather than uploadi ng your report as an 
attachment. Please only upload as attachments annotated versions of manuscripts, graphs, 
supporting materials or other aspects of your report which cannot be included in a text format.  
 
Please overwrite this text when adding your commen ts to the authors.  
 
Musashi1 predict poor prognosis and malignant transformation in hepatocellular carcinoma by 
activation of Wnt/β -catenin signaling pathway  
 
The authors looked at the Musashi1, which has noted to contribute to cancer progression in 
severa l cancers, in HCC.  
 
 
In the introduction  
 
Authors first showed that musashi1 was overexpressed in HCC tumors/cell lines compared to 
normal adjacent tissue. Figure 1A shows this finding. I suggest that authors quantify these 
findings and show them in a figu re in addition to what they showed (something similar to Fig 1 
C and D). In figure 1D, expression of musashi1 was increased in HCC cell lines compared to 
normal liver cells. P value should be reported.  
 
Later authors showed that musashi knockdown is associated with decreased cell viability and 
musashi overexpression is associated with increased viability. Two assays were used for this 
purpose which is good however we usually prefer to see this effect in m ore than two cells 
(usually three cell lines minimum specially that authors have four cell lines that showed 
overexpression of musashi1).  
 
The authors showed that the effect of musashi1 in HCC is through Wnt/β -catenin signaling 
pathway. This has been repo rted before. Chen K et. al. Liver Int. 2015 Mar;35(3):986 -98. doi: 
10.1111/liv.12458. Epub 2014 Jan 29.  
 
The authors showed that musashi1 overexpression is associated with poor prognosis of HCC 
patients. This is an interesting finding. Lu He et. al. showed  that musashi2, but not musashi1, 
was associated with poor prognosis. I think it worth discussing this contradictory finding in the 
current study. (J Cell Mol Med. 2014 Jan; 18(1): 49 -58). 
The manuscript is poorly written with numerous grammar mistakes (s ome sentences are not even 
understandable).  I will mention only few examples below.   
 
 
Examples:  
 
Page 2, line 51 -52: Authors mean to say PLC has become third cause of cancer related death 
worldwide however two mistakes are noted in this sentence: 1) the wo rd "which" is referring to 
morbidity and mortality in the way they stated it which is not what they mean. 2) They should 
add something like "cause/reason/etiology of" in between "third" and cancer -related death" 
otherwise the sentence doesn't make sense.  
 
Also in line 52: if primary liver cancer to be abbreviated as PLC, this abbreviation should be 
used later.   
 
Page 3, line 4 -5: authors started introducing Musashi1 gene and then they started describing the 
protein and stated "It has been identified as an e volutionarily conserved group of RNA binding 
protein and used as a molecular marker of neural stem cells."  
 
"It" is referring to the gene but they are describing the protein! This should be corrected.  
 
Page 3, line 7: "musashi" is spelled without "1"  
 
Page 6 , line 10: "were chose" should read were chosen.  
 
Throughout the manuscript: I got a sense that the authors naming the gene musashi1 and the 
protein musashi -1. If this is correct, those two words were used interchangeably throughout the 
manuscript i.e. sho uld be consistent.  
 
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the w ork 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.  
 
Yes 
 
Are yo u 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:  
 
Not suitable for publication unless extensively edited  
 
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 
publi cation 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 curr ently 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 understan d that my name will be included 
on my report to the authors and, if the manuscript is accepted for publication, my named repo rt 
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 understan d 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