The Potential Association of CDKN2A and Ki-67 Proteins in View of the Selected Characteristics of Patients with Head and Neck Squamous Cell Carcinoma

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Review due date was extended: October 20th, 2024

cimb-3256600, entitled “Potential association of CDKN2A and Ki-67 proteins in view of the selected characteristics of patients with head and neck squamous cell carcinoma” has been carefully reviewed.

 

Through a careful review of available literature, too many publications have reported the presentation of CDKN2A involved in HNSCC occurrence and development. I don’t see any novelty highlighted in this investigation. The conclusion has already been proved by other studies previously.

1. Riese U, Dahse R, Fiedler W, Theuer C, Koscielny S, Ernst G, Beleites E, Claussen U, von Eggeling F. Tumor suppressor gene p16 (CDKN2A) mutation status and promoter inactivation in head and neck cancer. Int J Mol Med. 1999 Jul;4(1):61-5. doi: 10.3892/ijmm.4.1.61. PMID: 10373639.

2. Calmon MF, Colombo J, Carvalho F, Souza FP, Filho JF, Fukuyama EE, Camargo AA, Caballero OL, Tajara EH, Cordeiro JA, Rahal P. Methylation profile of genes CDKN2A (p14 and p16), DAPK1, CDH1, and ADAM23 in head and neck cancer. Cancer Genet Cytogenet. 2007 Feb;173(1):31-7. doi: 10.1016/j.cancergencyto.2006.09.008. PMID: 17284367.

3. Kuroki M, Iinuma R, Okuda H, Terazawa K, Shibata H, Mori KI, Ohashi T, Makiyama A, Futamura M, Miyazaki T, Horikawa Y, Ogawa T. Comprehensive Genome profile testing in head and neck cancer. Auris Nasus Larynx. 2023 Dec;50(6):952-959. doi: 10.1016/j.anl.2023.04.006. Epub 2023 May 8. PMID: 37164815.

4. Shen A, Ye Y, Chen F, Xu Y, Zhang Z, Zhao Q, Zeng ZL. Integrated multi-omics analysis identifies CD73 as a prognostic biomarker and immunotherapy response predictor in head and neck squamous cell carcinoma. Front Immunol. 2022 Nov 16;13:969034. doi: 10.3389/fimmu.2022.969034. PMID: 36466881; PMCID: PMC9708745.

5. Deneka AY, Baca Y, Serebriiskii IG, Nicolas E, Parker MI, Nguyen TT, Xiu J, Korn WM, Demeure MJ, Wise-Draper T, Sukari A, Burtness B, Golemis EA. Association of TP53 and CDKN2A Mutation Profile with Tumor Mutation Burden in Head and Neck Cancer. Clin Cancer Res. 2022 May 2;28(9):1925-1937. doi: 10.1158/1078-0432.CCR-21-4316. PMID: 35491653; PMCID: PMC9186806.

6. Dragomir LP, Simionescu C, Mărgăritescu C, Stepan A, Dragomir IM, Popescu MR. P53, p16 and Ki67 immunoexpression in oral squamous carcinomas. Rom J Morphol Embryol. 2012;53(1):89-93. PMID: 22395505.

Author Response

Reviewer comment:

Through a careful review of available literature, too many publications have reported the presentation of CDKN2A involved in HNSCC occurrence and development. I don’t see any novelty highlighted in this investigation. The conclusion has already been proved by other studies previously.

1. Riese U, Dahse R, Fiedler W, Theuer C, Koscielny S, Ernst G, Beleites E, Claussen U, von Eggeling F. Tumor suppressor gene p16 (CDKN2A) mutation status and promoter inactivation in head and neck cancer. Int J Mol Med. 1999 Jul;4(1):61-5. doi: 10.3892/ijmm.4.1.61. PMID: 10373639.
2. Calmon MF, Colombo J, Carvalho F, Souza FP, Filho JF, Fukuyama EE, Camargo AA, Caballero OL, Tajara EH, Cordeiro JA, Rahal P. Methylation profile of genes CDKN2A (p14 and p16), DAPK1, CDH1, and ADAM23 in head and neck cancer. Cancer Genet Cytogenet. 2007 Feb;173(1):31-7. doi: 10.1016/j.cancergencyto.2006.09.008. PMID: 17284367.
3. Kuroki M, Iinuma R, Okuda H, Terazawa K, Shibata H, Mori KI, Ohashi T, Makiyama A, Futamura M, Miyazaki T, Horikawa Y, Ogawa T. Comprehensive Genome profile testing in head and neck cancer. Auris Nasus Larynx. 2023 Dec;50(6):952-959. doi: 10.1016/j.anl.2023.04.006. Epub 2023 May 8. PMID: 37164815.
4. Shen A, Ye Y, Chen F, Xu Y, Zhang Z, Zhao Q, Zeng ZL. Integrated multi-omics analysis identifies CD73 as a prognostic biomarker and immunotherapy response predictor in head and neck squamous cell carcinoma. Front Immunol. 2022 Nov 16;13:969034. doi: 10.3389/fimmu.2022.969034. PMID: 36466881; PMCID: PMC9708745.
5. Deneka AY, Baca Y, Serebriiskii IG, Nicolas E, Parker MI, Nguyen TT, Xiu J, Korn WM, Demeure MJ, Wise-Draper T, Sukari A, Burtness B, Golemis EA. Association of TP53 and CDKN2A Mutation Profile with Tumor Mutation Burden in Head and Neck Cancer. Clin Cancer Res. 2022 May 2;28(9):1925-1937. doi: 10.1158/1078-0432.CCR-21-4316. PMID: 35491653; PMCID: PMC9186806.
6. Dragomir LP, Simionescu C, Mărgăritescu C, Stepan A, Dragomir IM, Popescu MR. P53, p16 and Ki67 immunoexpression in oral squamous carcinomas. Rom J Morphol Embryol. 2012;53(1):89-93. PMID: 22395505.

Authors reponse:

Thank you for the comment. In the studies cited above, the authors focused on examining the methylation pattern of the CDKN2A gene by methylation-specific PCR (Riese et al.,1999; Calmon et al., 2007) and analyzing CDKN2A mutations through multi-omics and genomic data (Kuroki et al.,2023; Shen et al., 2022; Deneka et al., 2022), as well as p16 immunoexpression analysis (Dragomir et al., 2012). In our study, the methodology was quite different, as we analyzed CDKN2A and Ki-67 proteins by immunoenzymatic ELISA. The present study was designed to evaluate CDKN2A and Ki-67 concentrations and the possible association of clinical and demographic variables in tumors and resected surgical margin specimens from patients with primary HNSCC. In addition, our results are about the absence of statistical differences in CDKN2A and Ki-67 protein levels between tumor and HNSCC margins seem to be of interest, as we did not find similar studies analyzing CDKN2A and Ki-67 proteins by ELISA in tumor and surgical margin tissue homogenates.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Authors,

I read with great interest your article about possible associations between cdkn2a and ki-67 in HNSCC

However, there are some aspects requiring your attention.

Figure 1, please include the actual values in the chart.

Figure 2, please include the actual values in the chart.

Figure 3, please include the actual values in the chart.

Figure 4, please include the actual values in the chart.

Figure 5, please include the actual values in the chart.

Figure 6, please include the actual values in the chart.

There are many abbreviations in the text, please include a list of abbreviations at the end of the manuscript.

At the end of the discussion section insert a small paragraph about future directions of research. Such a future direction is the analysis of using other biomarkers such as molecular biology tests for further in-depth characterization of the tumors. Reference this to articles like doi: 10.3390/ijms25137350. PMID: 39000457; PMCID: PMC11242036.

Please update the references, there are titles from 1998 and 1999. I am sure that there are newer titles that are worth mentioning on the subject.

 

Looking forward to receiving the improved version of your manuscript.

Author Response

Reviewer:

Dear Authors,

I read with great interest your article about possible associations between cdkn2a and ki-67 in HNSCC

However, there are some aspects requiring your attention.

Figure 1, please include the actual values in the chart.

Figure 2, please include the actual values in the chart.

Figure 3, please include the actual values in the chart.

Figure 4, please include the actual values in the chart.

Figure 5, please include the actual values in the chart.

Figure 6, please include the actual values in the chart.

Authors: Thank you for your suggestion. We have added this information to the revised version of our manuscript, now every chart has added actual results and boxplots.

Reviewer:

There are many abbreviations in the text, please include a list of abbreviations at the end of the manuscript.

Authors: Thank you for your comment. We have added a list of abbreviations at the end of the article.

Reviewer:

At the end of the discussion section insert a small paragraph about future directions of research. Such a future direction is the analysis of using other biomarkers such as molecular biology tests for further in-depth characterization of the tumors. Reference this to articles like doi: 10.3390/ijms25137350. PMID: 39000457; PMCID: PMC11242036.

Authors: Thank you for your valuable consideration. We have added a short paragraph in the discussion regarding further research and included the reviewer's suggested publication.

Reviewer:

Please update the references, there are titles from 1998 and 1999. I am sure that there are newer titles that are worth mentioning on the subject.

Authors: Thank you for your suggestion. We have replaced older articles from 1999 and 1998 with newer publications.

Reviewer:

Looking forward to receiving the improved version of your manuscript.

Author Response File: Author Response.pdf