Metabolic Compartmentalization in Colorectal Cancer Hepatic Metastases and Correlation with Tumor Aggressiveness

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The presented manuscript concerns the role of MCT4, GLUT1, CD147, and CD44 in the pathology of CRC liver metastases. It briefly summarizes the aim of the study and is divided into individual sections in which the authors accurately explain the carried out research. In terms of content, the information was presented fairly and accurately. The work is a valuable report, however, some clarification is necessary.

1. According to the manuscript, the study group was gathered in 2003-2010, but the data collection and usage were approved in June 2011. Has consent to collect the material been previously given? Additionally, the "Institutional Review Board Statement" section contain consents from 2013. This requires explanation.

2. Were the tests performed during the consent of the bioethics committee? If so, approximately in what period? In the authors' opinion, is the material stored for such a long time suitable for use? About 20 years have passed since it was collected, which concerns me.

3. The authors have written: "The absence of correlations between the expression of the other markers and the clinicopathological data could be due to the small sample size of the study." That is true, but maybe it can be also connected with such a long storage of material used?

4. The authors wrote: "The findings suggest critical roles for MCT4, GLUT1, CD147, and CD44 in the pathology of liver metastases from CRC, providing insights into potential therapeutic targets." Based on the analysis of the presented results, I believe that this sentence is exaggerated.

5. An additional section showing the limitations of the study would be useful.

Comments on the Quality of English Language

The paper is written in a thoughtful and understandable way.

Author Response

Thank you very much for your comments.

We appreciate the comments and suggestions that allow us to improve the manuscript.

Here are the answers:

 

1.According to the manuscript, the study group was gathered in 2003-2010, but the data collection and usage were approved in June 2011. Has consent to collect the material been previously given? Additionally, the "Institutional Review Board Statement" section contain consents from 2013. This requires explanation.

Answer:  All operated patients, with surgical specimens analyzed by the department of pathological anatomy, are left with paraffin blocks in this service.

2. Were the tests performed during the consent of the bioethics committee?If so, approximately in what period? In the authors' opinion, is the material stored for such a long time suitable for use? About 20 years have passed since it was collected, which concerns me.

Answer:  Yes, Ethic  Committee approved the work. The paraffin block are stored for long period for reanalyzing it when necessary, the TMA blocks are performed when the study was approved by Ethic Committee

3. The authors have written: "The absence of correlations between the expression of the other markers and the clinicopathological data could be due to the small sample size of the study." That is true, but maybe it can be also connected with such a long storage of material used?

Answer:  The samples were preserved in paraffin-embedded preparation. The small sample is regarded to the size of the tissue analyzed.

4. The authors wrote: "The findings suggest critical roles for MCT4, GLUT1, CD147, and CD44 in the pathology of liver metastases from CRC, providing insights into potential therapeutic targets." Based on the analysis of the presented results, I believe that this sentence is exaggerated.

Answer:  We rephrased the sentence for: The findings for MCT4, GLUT1, CD147, and CD44 obtained in this study are very promising to consider these markers as therapeutic targets in further investigations. (yellow)

5. An additional section showing the limitations of the study would be useful.

Answer:  The study has an imitation regarding the amplitude of the tissue reserche due to the limited size of the samples, which prevent a more accurate analyzis of the cytological compartments of the tumor cells and the surrounding tissues. (yellow)

 

Reviewer 2 Report

Comments and Suggestions for Authors

The authors present a study of the inmunohistochemistry staining of MCT4, GLUT1, CD147 and CD44 in various cellular and histological compartments of liver metastases from Colorectal Cancer samples. The manuscript is carefullly writen and presented. The Introduction is correct and describes the interest of the objective of the study.

But there are some points that need to be improved.

- The authors have analized the inmunoexpression of the proteins in tissues from the metastasis. But not in tisssue samples from the primary tumors. The study would be much more interesting if they could analize also tissues from the primary tumors and compare the results from these two sites.

- The authors mention several times through the text that have analized the relation between inmunostaining of GLUT1 and CEA levels. Like in "Line 75" (... CEA levels at primary tumor and liver metastasis diagnosis, presence of extrahepatic metastases, adyuvant therapy,... ).   But it not clearly mentioned that CEA values correspond to serum levels and not tissue expression. It should be clearly defined.

- Discussion must be improved. Discuss the trascendence of the results obtained. Compare the results with those from other publications. Comment which may be the role of the four markers studied in the process of tumor progression and metastasis 

 

Comments on the Quality of English Language

English language is correct

Author Response

Thank you very much for your comments.

We appreciate the comments and suggestions that allow us to improve the manuscript.

Here are the answers:

1.The authors have analized the inmunoexpression of the proteins in tissues from the metastasis. But not in tisssue samples from the primary tumors. The study would be much more interesting if they could analize also tissues from the primary tumors and compare the results from these two sites.

Answer: The immunoexpression of the primary tumors were analyzed previously as mentioned in the text Reference 16, in other different analyzes. (16.           (Martins, S.F.; Martins, S. F., Amorim, R., Viana-Pereira, M., Pinheiro, C., Costa, R. F. A., Silva, P., Couto, C.; Alves, S.; Fernandes, S.; Vilaça, S.; et al. Significance of glycolytic metabolism-related protein expression in colorectal cancer, lymph node and hepatic metastasis. BMC Cancer 2016, 16, 535. DOI: 10.1186/s12885-016-2566-9).

- The authors mention several times through the text that have analized the relation between inmunostaining of GLUT1 and CEA levels. Like in "Line 75" (... CEA levels at primary tumor and liver metastasis diagnosis, presence of extrahepatic metastases, adyuvant therapy,... ).   But it not clearly mentioned that CEA values correspond to serum levels and not tissue expression. It should be clearly defined.

Answer:  CEA values correspond to serum levels and not tissue expression. We add this information to the materials and methods (in red …line 73)

- Discussion must be improved. Discuss the trascendence of the results obtained. Compare the results with those from other publications. Comment which may be the role of the four markers studied in the process of tumor progression and metastasis 

Answer:  We've added data to the discussion... in red

 

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The authors responded to all objections, therefore the work can be accepted in its current form.

Comments on the Quality of English Language

The paper is written in a thoughtful and understandable way.

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript has been sufficiently improved to be published

Comments on the Quality of English Language

The english language needs minor revision