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Updated: Oct 2, 2025
Rapid Generation of Primary Murine Melanocyte and Fibroblast Cultures
Published on: June 26, 2019
Establishment and characterization of an immortalized human giant congenital melanocytic nevi cell line
Qingxiong Yu1, Rehanguli Aimaier1, Man-Hon Chung1
1Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, People's Republic of China.
Abstract:
Treatments for giant congenital melanocytic nevi (GCMN) are extremely limited. Thus, there is an urgent need for development of relevant targeted therapies. However, current lack of preclinical cell models restricts progress in GCMN research. In this study, we aimed to establish and characterize an immortalized GCMN cell line. GCMN cells were successfully immortalized by means of lentivirus-mediated simian virus 40 large T transfection. The immortalized GNC cell line (ImGNC) showed lower proliferation rate and higher melanin content than primary melanocytes. Expression levels of the differentiation gene MITF and stemness genes TWIST1, SNAI1, and FOXD3 were elevated in ImGNCs; however, the established ImGNC cell line was immortalized but not transformed. Sanger sequencing detected the heterozygous NRASQ61K mutation in ImGNCs, but not the BRAFV600E mutation. Despite carrying the NRASQ61K allele, ImGNCs demonstrated suppressed MAPK activation and elevated PI3K/Akt activation, as compared with primary melanocytes. Drug sensitivity analysis showed that ImGNCs are more sensitive to PI3K/Akt and Bcl-2 inhibitors than to MEK or ERK inhibitors. Unlike the proliferation-inhibiting effect of PI3K/Akt inhibitors, the Bcl-2 inhibitor navitoclax promptly promoted apoptosis in ImGNCs. Considering the low proliferation characteristics of GCMN in vivo, Bcl-2 may be a potential therapeutic target that warrants further research.
Insights
Researchers developed an immortalized giant congenital melanocytic nevi (GCMN) cell line. This new GCMN cell model shows sensitivity to PI3K/Akt and Bcl-2 inhibitors, suggesting potential therapeutic targets.
Area of Science:
- Dermatology
- Oncology
- Cell Biology
Background:
- Treatments for giant congenital melanocytic nevi (GCMN) are limited, necessitating novel therapeutic strategies.
- A lack of suitable preclinical models hinders research and drug development for GCMN.
- Establishing a reliable GCMN cell line is crucial for advancing understanding and treatment.
Purpose of the Study:
- To establish and characterize an immortalized GCMN cell line for preclinical research.
- To investigate the molecular characteristics and drug sensitivities of the new cell line.
- To identify potential therapeutic targets for GCMN.
Main Methods:
- Immortalization of GCMN cells using lentivirus-mediated simian virus 40 large T transfection.
- Characterization of the immortalized cell line (ImGNC) for proliferation, melanin content, gene expression, and mutational status.
- Analysis of signaling pathway activation (MAPK, PI3K/Akt) and drug sensitivity (PI3K/Akt, Bcl-2, MEK, ERK inhibitors).
Main Results:
- Successfully established an immortalized GCMN cell line (ImGNC) with lower proliferation and higher melanin content than primary melanocytes.
- ImGNC cells exhibited elevated MITF, TWIST1, SNAI1, and FOXD3 expression and carried an NRASQ61K mutation.
- ImGNCs showed suppressed MAPK and elevated PI3K/Akt activation, with increased sensitivity to PI3K/Akt and Bcl-2 inhibitors, particularly navitoclax-induced apoptosis.
Conclusions:
- The novel ImGNC cell line serves as a valuable preclinical model for GCMN research.
- Targeting Bcl-2 is a promising therapeutic strategy for GCMN, given its role in inducing apoptosis in this cell line.
- Further investigation into Bcl-2 as a therapeutic target for GCMN is warranted.
