Differential Gene Expression and Tumorigenicity Analysis of Cultured Melanocyte Comparing Melanoma

Shahbazi, Atefeh; Zargar, Seyed Jalal; Bajouri, Amir; Mohammadi, Parvaneh; Aghdami, Nasser

doi:10.22088/IJMCM.BUMS.13.4.387

Volume 13, Issue 4 (Int J Mol Cell Med 2024) Int J Mol Cell Med 2024, 13(4): 387-403 | Back to browse issues page

‎ 10.22088/IJMCM.BUMS.13.4.387

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Shahbazi A, Zargar S J, Bajouri A, Mohammadi P, Aghdami N. Differential Gene Expression and Tumorigenicity Analysis of Cultured Melanocyte Comparing Melanoma. Int J Mol Cell Med 2024; 13 (4) :387-403
URL: http://ijmcmed.org/article-1-2254-en.html

Differential Gene Expression and Tumorigenicity Analysis of Cultured Melanocyte Comparing Melanoma

Atefeh Shahbazi¹

, Seyed Jalal Zargar²

, Amir Bajouri³

, Parvaneh Mohammadi⁴

, Nasser Aghdami⁴

1- Department of Cellular and Molecular Biology, Kish International Campus, University of Tehran, Tehran, Iran.
2- Department of Cell and Molecular Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran. , zargar@ut.ac.ir
3- Skin & Stem cell Research Center, Tehran University of Medical Sciences, Tehran, Iran.
4- Department of Regenerative Medicine, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.

Abstract: (618 Views)

This study aimed to identify the optimal growth media for culturing human skin melanocytes for clinical applications and to assess their tumorigenic potential both in vitro and in vivo. Various growth media were tested to determine the most effective and safest for melanocyte culture, avoiding harmful growth factors such as TPA and colorant toxins. The study evaluated changes in RAF and NRAS gene expression through real-time PCR and gene sequencing of BRAF V600E and NRAS in exons 1 and 2, comparing these with melanoma. Melanocytes were subcutaneously injected into BALB/c nude mice to assess tumorigenic risk. Results indicated that a mixture of MGM-M2 supplemented with melanocyte growth factors provided the best outcomes in terms of cell proliferation and melanocyte count. Gene expression analysis revealed that HRAS and BRAF expressions in melanocytes at passage 6 showed less than 2-fold increases, whereas these genes were up-regulated by more than 3 and 8 folds, respectively, in melanoma cell lines. NRAS expression in melanocytes at passage 6 increased by 5-fold but remained lower than in melanoma cell lines. Gene sequencing of BRAF V600E and NRAS in exons 1 and 2 showed no mutations, and melanocytes injected into BALB/c nude mice exhibited no tumor formation risk. Furthermore, gene sequencing of BRAF and NRAS in the injected melanocytes 16 weeks’ post-transplantation revealed no mutations. These findings suggest that while standard growth media protocols may elevate specific proto-oncogene expressions, they do not induce tumorigenic mutations in melanocytes, both in vitro and in vivo.

Keywords: Cultured melanocyte, Vitiligo, cell transplantation, tumorigenicity

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Type of Study: Original Article | Subject: Stem Cell
Received: 2023/12/3 | Accepted: 2024/08/27 | Published: 2024/11/30