Effect of gedunin on cell proliferation and apoptosis in skin melanoma cells A431 via the PI3K/JNK signaling pathway

Adv Clin Exp Med. 2024 Nov 12. doi: 10.17219/acem/189729. Online ahead of print.

Abstract

Background: Melanoma is an aggressive skin malignancy with rapid metastasis and high morbidity. Gedunin (GN) is a tetranortriterpenoid belonging to the Meliaceae family, described for its anticancer, antiproliferative and apoptotic properties.

Objectives: In the present study, we investigated the effect of GN on A431 melanoma cell proliferation and apoptosis. The inflammatory proteins (tumor necrosis factor alpha (TNF-α), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), cycloxygenase 2 (COX-2), inducible nitric oxide synthase (iNOS), and interleukin 6 (IL-6)) apoptosis-related proteins, such as Bax, Bcl-2 and caspase-3, and alterations in the PI3K/JNK and p38 pathways in A431 cells after GN treatment were examined.

Material and methods: The cytotoxicity assay and cell apoptosis of GN activity on A431 cells were assessed using MTT assay, acridine orange/ethidium bromide (AO/EB), DAPI (4',6-diamidino-2-phenylindole), propidium iodide (PI), enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR) and western blot analyses.

Results: The findings demonstrated that GN (10 and 15 μM/mL) inhibits the growth of melanoma cells, triggers apoptosis by enhancing Bax and caspase, and reduces Bcl-2, cyclin-D1, c-myc, and survivin in a concentration-reliant manner. Additionally, GN attenuated the protein expression of inflammatory proteins (TNF-α, NF-κB, COX-2, iNOS, and IL-6) and the cell proliferative PI3K/JNK/p38 signaling pathway. Due to the imbalance in the Bax/Bcl-2 ratio, apoptosis is promoted, and the caspase cascade and Cyt-c are activated. This led us to conclude that GN treatment inhibited Bcl-2, cyclin-D1, c-myc, and survivin activity through the TNF-α/NF-κB and PI3K/JNK/p38 signaling pathways, further preventing the proliferation and stimulation of apoptosis, which contributes to growth arrest in melanoma cells.

Conclusions: Gedunin has been shown to promote melanoma cell death in vitro, suggesting that it could be used as a future treatment for malignant melanoma. Our findings suggested that GN might be applied as a preventative measure in the management of skin melanoma cells.

Keywords: AROS; P13K/JNK/p38 signaling; apoptosis; gedunin; skin cancer.