Human Metastatic Melanoma Cell Lines Panel for In Vitro and In Vivo Investigations
, Nadezhda A. Kalinina 1, Ksenia M. Konoplina 1, Anastasiia A. Malchenkova 1, Alexandra E. Evdokimova 1, Marina V. Piniugina 1, Irina I. Khan 1, Ilya A. Kislyak 2, Anna A. Basharina 1, Anna N. Grishanina 1, Anna A. Rudakova 1, Pavel O. Varaksa 1, Maria A. Baryshnikova 1, Vadim S. Pokrovsky 1,2,*, Tatiana A. Bogush 1 and Vyacheslav S. Kosorukov 1,*Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe authors developed melanoma cell lines from metastatic tumors in three patients, all originating from lymph node metastases. They characterized these cell lines using various methods.
However, the title, "Human melanoma cell lines panel for in vitro and in vivo investigations," could be misleading, as it implies a more extensive panel of cell lines from various primary tumors and metastatic sites. To make it clear, the title should be corrected to specify that all the cell lines are from lymph node metastases.
The abstract is missing crucial information about the origin of the cell lines, making it difficult to understand how the cell line names were chosen. I suggest including information on the criteria used for naming the cell lines (e.g. in the Materials and Methods)
Why is it important to note in the abstract that TUBB3 were expressed in all 3 cell lines? What is the importance of TUBB3 in metastatic melanoma? In addition, it's important to correct the gene name to 'TUBB3'.
The names of the genes should be written in italic.
I do not understand what the authors meant as a conclusion at the end of the abstract: "The multiparametric characterization of the cell lines makes this panel a valuable product for scientific and regulatory experiments aimed to obtain non-clinical data on the antiproliferative activity of new agents for malignant melanoma and/or investigation of melanoma cells properties and progression."
The numbers in the introduction are not properly written, it should be corrected.
The conclusion in the abstract is somewhat unclear: “The multiparametric characterization of the cell lines makes this panel a valuable product for scientific and regulatory experiments aimed to obtain non-clinical data on the antiproliferative activity of new agents for malignant melanoma and/or investigation of melanoma cells properties and progression."; It appears to suggest that the detailed characterization of the cell lines makes them valuable for various scientific and regulatory experiments, particularly those involving non-clinical data on anti-proliferative agents for malignant melanoma or investigations into melanoma cell properties and progression. However, the statement could be more precise in explaining the specific applications and benefits of this panel for scientific research and regulatory purposes.
The introduction has several shortcomings: it should include some of Menhard Herlyn's papers, his laboratory has the largest collection of melanoma cell lines.
Important references appear to be missing from the introduction. It's noteworthy that no references are cited from line 37 onwards in the introduction. To provide a robust foundation for the research, please ensure that relevant references are cited and integrated throughout the introduction.
Materials and Methods: Thee origin of tumours are missing (primary melanoma? melanoma metastasis?)
The number and dates of the approval by the ethics committee for the study is missing: Lines 54-55.
It would be interesting to know what were the primary melanoma clinoco-pathological parameters (histological subtypes, Breslow thickness, presence of ulceration etc.)
The quality of Figure 2. is poor.
Melanoma tumours are very heterogeneous, it is questionable whether the karyotypes on Figure 3. are representative or not? This should be stated.
Figure 4. poor quality and this is true for all figures.
Line 407 as far as I know CDKN2A is one of the most studied tumor suppressor gene, not an oncogene.
Missing Statements:
Institutional Review Board Statement: Line 429
Informed Consent Statement: Line 430
Data Availability Statement: Line 431
Supplementary Tables, Figures are not indicated in the main text. Is there any video uploaded (Line 425)?
Supplementary Figures/Tables: Table S1: all numbers should be corrected.
Table S2: should be corrected, part of the Table is missing, numbers are not correctly written
Figure S1: Figure legend is not informative, the quality of the figures are poor.
Author Response
Please see the attachment.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for Authors
The authors isolated new primary cell lines for the analysis of melanomas with special focus to verify therapy options. The goal is to generate newly isolated material for experiments in the laboratory.
The reviewer feels that the Introduction is too short. There are no arguments why specific marker genes (PD-L1, ERa, CK) were selected, nor why MC1R-TUBB3 was given such a high priority. Why do the authors do not perfom PCR analysis to discriminate TUBB3 and the splice variant MC1R-TUBB3?
The special splicing of TUBB3 is not described at all in the Introduction:
Human MC1R has an inefficient poly(A) site allowing intergenic splicing with its downstream neighbour Tubulin-β-III (TUBB3). Intergenic splicing produces two MC1R isoforms, designated Iso1 and Iso2, bearing the complete seven transmembrane helices from MC1R fused to TUBB3-derived C-terminal extensions (Herraiz et al., 2015).
The reviewer is interested in whether it will be possible for researchers worldwide to access the new cell lines? How open is the team for cooperation regarding the cell lines? There is no statement at the end of the manuscript.
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Comments and Suggestions for AuthorsTitle: the suggested title: Establishement and characterization of human metastatic melanoma cell lines panel for in vitro and in vivo investigations
Abstract: still missing the origin/the metastatic origin of the cell lines.
TUBB3 should be written consistently, sometimes it is written as TUBB 3
Introduction: The literature citations for lines 38-43 are still missing. At least the latest review should be cited for new agents for melanoma treatment.
The references for the new part of the introduction (from line 44) are not really correct e.g. for Herlyn's work it should be number 3 not number 4 ... please revise this part.
Figure 3. title should be corrected: Representative karyotypes of melanoma cell lines.
Author Response
1. Title: the suggested title: Establishement and characterization of human metastatic melanoma cell lines panel for in vitro and in vivo investigations
We can agree with this title.
2. Abstract: still missing the origin/the metastatic origin of the cell lines.
Addition included in abstract.
3. TUBB3 should be written consistently, sometimes it is written as TUBB 3
Inaccuracies have been corrected.
4. Introduction: The literature citations for lines 38-43 are still missing. At least the latest review should be cited for new agents for melanoma treatment.
Links added.
5. The references for the new part of the introduction (from line 44) are not really correct e.g. for Herlyn's work it should be number 3 not number 4 ... please revise this part.
We double-checked, everything is correct.
6. Figure 3. title should be corrected: Representative karyotypes of melanoma cell lines.
