Title: Tumor microenvironment-related gene selenium-binding protein 1 (SELENBP1) is
associated with immunotherapy efficacy and survival in colorectal cancer
Version: 1 Date: 09 Mar 2022
Reviewer's report:
Major concerns:
1) The authors depict in abstract: “..it is currently lacking research about relationship between
SELENBP1 and immunotherapy in colorectal cancer (CRC).” However, I found no particular
introduction/further explanation of such statement in introduction section. Please, describe more,
why such relationship was evaluated in the present study. What is known previously?
2) 2.10 and 2.11 sections are very poorly described. What antibodies (against what, dilutions)
were used in the present study? What chemiluminescence assay was applied? Please specify
more carefully. Next, in 189/190 lines: the authors declare “Sections were incubated with
SELEBNP1 (1:200 diluted), visualized with DAB detection.” Did the authors incubated the
histology specimens with SELEBNP1 protein itself? It doesn’t make any sense.. The information
about providers is also missing.
Significant traces of DNA are observed during RNA extraction using TRISOL method. To prove
that qPCR evaluation is not biased by DNA contamination, please provide additional qPCR data
from samples treated with Dnase.
3) From 3.1 and figure 1A perspective, why further exploration on CRC was selected? The
authors show a significant difference between tumor and normal tissue in 27 types of cancers..
Please describe clearly.
4) It is unclear what is shown in figure 1 D-I. Please change figure style.
5) Figures in figure 1 K and L are taken directly from HPA. I don’t see any results here obtained
by the authors. Please remove.
6) Line 243: the authors depict a negative correlation between SELENBP1 and invasion. Please
specify statistical values. It seems that correlation value is very week and insignificant from
figure 3d.
7) Line 251: please describe results of heat map in more detail. It is not clear, what does it mean..
8) In sections 3.5-3.8, the presentation of the results could be significantly organized in better
way. For instance, where are much graphic material with insignificant data (low correlation
coeficients etc.). Much data could be represented as supplementary material. It is hard to absorb
such load of characteristics without clear description in the manuscript text.
9) The manuscript results are extensively based on bioinformatic analysis, please provide R code
in github for the analysis.
Minor concerns:
1) In 44 row: please change English style:
” We found that SELENBP1 is lowly expressed in CRC and is associated with poor prognosis.”
to „We found that low expression of SELENBP1 in CRC (compared to what? Adjacent tissue?)
was associated with poor prognosis.
2) In 58 row: “These studies suggest that SELENBP1 can be considered as a promising
prognostic biomarker for CRC. SELENBP1..” What studies? Please specify.
3) In 76 row: „SELENBP1 is a selenium containing protein that binds selenium atoms in
cytoplasm and nucleus.” Does SELENBP1 contain selenium (selenium methionine) itself, also
bind selenium or both? Please specify.
4) In figure 1, Meso and Uvm don’t have SELENBP1 expression from normal  tissue. Please
remove.
Title: Tumor microenvironment-related gene selenium-binding protein 1 (SELENBP1) is
associated with immunotherapy efficacy and survival in colorectal cancer
Version: 1 Date: 09 Mar 2022
Reviewer's report:
Major concerns:
1) The authors depict in abstract: “..it is currently lacking research about relationship between
SELENBP1 and immunotherapy in colorectal cancer (CRC).” However, I found no particular
introduction/further explanation of such statement in introduction section. Please, describe more,
why such relationship was evaluated in the present study. What is known previously?
2) 2.10 and 2.11 sections are very poorly described. What antibodies (against what, dilutions)
were used in the present study? What chemiluminescence assay was applied? Please specify
more carefully. Next, in 189/190 lines: the authors declare “Sections were incubated with
SELEBNP1 (1:200 diluted), visualized with DAB detection.” Did the authors incubated the
histology specimens with SELEBNP1 protein itself? It doesn’t make any sense.. The information
about providers is also missing.
Significant traces of DNA are observed during RNA extraction using TRISOL method. To prove
that qPCR evaluation is not biased by DNA contamination, please provide additional qPCR data
from samples treated with Dnase.
3) From 3.1 and figure 1A perspective, why further exploration on CRC was selected? The
authors show a significant difference between tumor and normal tissue in 27 types of cancers..
Please describe clearly.
4) It is unclear what is shown in figure 1 D-I. Please change figure style.
5) Figures in figure 1 K and L are taken directly from HPA. I don’t see any results here obtained
by the authors. Please remove.
6) Line 243: the authors depict a negative correlation between SELENBP1 and invasion. Please
specify statistical values. It seems that correlation value is very week and insignificant from
figure 3d.
7) Line 251: please describe results of heat map in more detail. It is not clear, what does it mean..
8) In sections 3.5-3.8, the presentation of the results could be significantly organized in better
way. For instance, where are much graphic material with insignificant data (low correlation
coeficients etc.). Much data could be represented as supplementary material. It is hard to absorb
such load of characteristics without clear description in the manuscript text.
9) The manuscript results are extensively based on bioinformatic analysis, please provide R code
in github for the analysis.
Minor concerns:
1) In 44 row: please change English style:
” We found that SELENBP1 is lowly expressed in CRC and is associated with poor prognosis.”
to „We found that low expression of SELENBP1 in CRC (compared to what? Adjacent tissue?)
was associated with poor prognosis.
2) In 58 row: “These studies suggest that SELENBP1 can be considered as a promising
prognostic biomarker for CRC. SELENBP1..” What studies? Please specify.
3) In 76 row: „SELENBP1 is a selenium containing protein that binds selenium atoms in
cytoplasm and nucleus.” Does SELENBP1 contain selenium (selenium methionine) itself, also
bind selenium or both? Please specify.
4) In figure 1, Meso and Uvm don’t have SELENBP1 expression from normal  tissue. Please
remove.
Please review this paper.
user
Title: Tumor microenvironment-related gene selenium-binding protein 1 (SELENBP1) is
associated with immunotherapy efficacy and survival in colorectal cancer
Version: 1 Date: 09 Mar 2022
Reviewer's report:
Major concerns:
1) The authors depict in abstract: “..it is currently lacking research about relationship between
SELENBP1 and immunotherapy in colorectal cancer (CRC).” However, I found no particular
introduction/further explanation of such statement in introduction section. Please, describe more,
why such relationship was evaluated in the present study. What is known previously?
2) 2.10 and 2.11 sections are very poorly described. What antibodies (against what, dilutions)
were used in the present study? What chemiluminescence assay was applied? Please specify
more carefully. Next, in 189/190 lines: the authors declare “Sections were incubated with
SELEBNP1 (1:200 diluted), visualized with DAB detection.” Did the authors incubated the
histology specimens with SELEBNP1 protein itself? It doesn’t make any sense.. The information
about providers is also missing.
Significant traces of DNA are observed during RNA extraction using TRISOL method. To prove
that qPCR evaluation is not biased by DNA contamination, please provide additional qPCR data
from samples treated with Dnase.
3) From 3.1 and figure 1A perspective, why further exploration on CRC was selected? The
authors show a significant difference between tumor and normal tissue in 27 types of cancers..
Please describe clearly.
4) It is unclear what is shown in figure 1 D-I. Please change figure style.
5) Figures in figure 1 K and L are taken directly from HPA. I don’t see any results here obtained
by the authors. Please remove.
6) Line 243: the authors depict a negative correlation between SELENBP1 and invasion. Please
specify statistical values. It seems that correlation value is very week and insignificant from
figure 3d.
7) Line 251: please describe results of heat map in more detail. It is not clear, what does it mean..
8) In sections 3.5-3.8, the presentation of the results could be significantly organized in better
way. For instance, where are much graphic material with insignificant data (low correlation
coeficients etc.). Much data could be represented as supplementary material. It is hard to absorb
such load of characteristics without clear description in the manuscript text.
9) The manuscript results are extensively based on bioinformatic analysis, please provide R code
in github for the analysis.
Minor concerns:
1) In 44 row: please change English style:
” We found that SELENBP1 is lowly expressed in CRC and is associated with poor prognosis.”
to „We found that low expression of SELENBP1 in CRC (compared to what? Adjacent tissue?)
was associated with poor prognosis.
2) In 58 row: “These studies suggest that SELENBP1 can be considered as a promising
prognostic biomarker for CRC. SELENBP1..” What studies? Please specify.
3) In 76 row: „SELENBP1 is a selenium containing protein that binds selenium atoms in
cytoplasm and nucleus.” Does SELENBP1 contain selenium (selenium methionine) itself, also
bind selenium or both? Please specify.
4) In figure 1, Meso and Uvm don’t have SELENBP1 expression from normal  tissue. Please
remove.
assistant