Review for 10.1186/s12876-023-02815-2 (2023)

Reviewer’s report
Title:  Glucose metabolic upregulation via phosphorylation of S6 ribosomal protein affects tumor
progression in distal cholangiocarcinoma
Version: 1 Date: 26 Mar 2023
Reviewer's report:
In the present manuscript, the authors demonstrated the possible role of phosphorylated S6 ribosomal
protein (pS6) on the glucose metabolism in distal cholangiocarcinoma (dCCA). The expression and
clinical correlation of pS6/GLUT1 in dCCA were determined using IHC technique. In addition, the
effect of pS6 on glucose metabolism in CCA cell lines was also investigated using an inhibitor of S6
phosphorylation  namely,  PF-04691502.  The  experiments  were  performed  in  a  technically  sound
manner. However, the following points need to be addressed before publication. 
Major points:
According to the following references as shown below, there is no distal cholangiocarcinoma was used 
in the present study. MEC, TFK-1, HuCCT1, and TKKK cell lines were established from non-distal 
cholangiocarcinoma. How can we verify or ensure that the findings from non-distal 
cholangiocarcinoma cell lines are applicable to distal cholangiocarcinoma? Please add more 
information about this point in the discussion session. 
1.      Yoshida K., Tomizawa H., Ota T., Nagashima T., Kikuchi H., Watanabe H., Hashizaki K., Yonaha 
A. Establishment and characterization of human cholangiocarcinoma, MEC, producing carbohydrate 
antigen 19-9. Hum. Cell 3:346-351(1990)
2.      Saijyo S., Kudo T., Suzuki M., Katayose Y ., Shinoda M., Muto T., Fukuhara K., Suzuki T., 
Matsuno S. Establishment of a new extrahepatic bile duct carcinoma cell line, TFK-1. Tohoku J. Exp. 
Med. 177:61-71(1995)
3.      Miyagiwa M., Ichida T., Tokiwa T., Sato J., Sasaki H. A new human cholangiocellular carcinoma 
cell line (HuCC-T1) producing carbohydrate antigen 19/9 in serum-free medium. In Vitro Cell. Dev. 
Biol. 25:503-510(1989)
4.      Sato J., Kimura T., Saito T., Anazawa T., Kenjo A., Sato Y., Tsuchiya T., Gotoh M. Gene 
expression analysis for predicting gemcitabine resistance in human cholangiocarcinoma. J. 
Hepatobiliary Pancreat. Sci. 18:700-711(2011) 
Declaration of competing interests
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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?
Yes
Is the presentation of the work clear?
Yes
Are  the  images  in  this  manuscript  (including  electrophoretic  gels  and  blots)  free  from  apparent
manipulation?
Yes