The Expression of Testin, Ki-67 and p16 in Cervical Cancer Diagnostics
, Jedrzej Grzegrzolka 1, Aleksandra Piotrowska 1, Mateusz Olbromski 1, Beata Smolarz 2, Hanna Romanowicz 2, Agnieszka Rusak 1, Monika Mrozowska 1, Piotr Dziegiel 1,3, Marzenna Podhorska-Okolow 4 and Christopher Kobierzycki 1Round 1
Reviewer 1 Report
In the presented manuscript, the authors investigate the immunohistochemical (ICH) expression of the testin protein in samples from cervical cancer patients, in relation to Ki-67 antigen and p16 protein. The authors propose the combined investigation of these three markers to improve diagnostic accuracy in cervical cancer diagnosis.
Although very limited, as also pointed out in the limitation section, this work reveal interesting directions for further research. Nonetheless, some issues need to be addressed to strengthen the results.
1) In figure 1, please add Spearman correlation coefficient on graphs, to better appreciate the presence or absence of correlation between tested variables.
2) The results section heading is missing
3) Bands quantification of western-blot reported in figures 3 and 4 should be added.
4) Figure 3 and 4 should be reported as unique figure, and molecular weights should also be indicated.
5) The authors proved that testin expression is higher in control cells, whereas it is weaker in cancer cell lines and almost not appreciable in HPV-negative cells. It is important to point out that these results have been obtained by Western-blot analysis, which evaluates the presence or absence of the protein within the cell. To complete the investigation, authors should perform RT-PCR on the different cell lines tested in this paper, so to provide a complete picture of testin expression.
6) Western-blot analysis, and relative RT-PCR, should be extended also to Ki-67 and p16, to have a more accurate insight into the relationships among the three agents.
Author Response
Please see the attachment.
Author Response File: 
Author Response.docx
Reviewer 2 Report
After carefully reading the paper, some corrections are mandatory.
1.Each statement of fact and each methd should be supported by References. All References should be recent, relevant, and accessible to readers of this journal.
2. Throughout the manuscript, please define all abbreviations when first used in the Abstract, the main manuscript, in each Table footnote, and in each Figure legend so that the manuscript content can be understood.
3. Publications are searched and cited based on the content of the manuscript Title, Abstract, and Keywords. Therefore, a clear and descriptive Title is required.
4. In the Results subsection of the Abstract, please add the p-values to show the significance of the main comparative results.
5. The Conclusions subsection of the Abstract should clearly and concisely state the main message from your study.
6. Please include a maximum of 5 Keywords using terms from the Medical Subject Heading (MeSH) database on the PubMed site (http://www.ncbi.nlm.nih.gov/mesh).
7. In the main Background section, please support each statement of fact with a Reference citation.
8. Please begin the main Methods section with the Ethical statement that includes the Ethics Committee approval details and also the details of patient informed consent.
9. The Discussion section must end with a paragraph discussing the study's limitations. Also, please discuss the limitations of the main methods
10. The main Conclusions section should be clear and concise and match the Abstract.
11. Please clarify how patients’ data was protected.
12. Please provide the name of the test used to analyze the normal distribution of obtained data.
13. In my option, the title of this sub-section is not properly "Tissue microarrays (TMAs) "
14. Each table/figure has to be self-explanatory. It is lacking in this paper.
Author Response
Please see the attachment.
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
In my opinion, RT-PCR data would strengthen the presented results, therefore I strongly suggest authors to add it here. For this reason, I think a minor revision is still needed.
Author Response
Thank you for your valuable suggestions. As this study is a part of bigger research project for sure we will take into account remarks of the reviewer. We are thankful for all amendments which improve quality of the paper. At this point we will not be able to extend technical issues. We are happy that we addressed all other points of the reviewer opinion. Best regards and all the best in 2023.
