Impact of trehalose and hydroxychloroquine on the function of BRAF (V۶۰۰E)-siRNA in the A۳۷۵ melanoma cells

Objective(s): Melanoma, a lethal form of skin cancer, is closely linked to mutations in melanocytes. Due to increased resistance to chemotherapy, innovative strategies, including gene and combination therapies, are being explored. This study evaluates the effects of trehalose (TRE) and hydroxychloroquine (HCQ) in enhancing the efficacy of BRAF (V۶۰۰E)-siRNA in A۳۷۵ cells.Materials and Methods: The A۳۷۵ cells were treated, and changes in cell viability were assessed using MTT assays. Apoptosis was evaluated using flow cytometry. Additionally, gene expression analysis of B-Raf proto-oncogene, serine/threonine kinase (BRAF), Caspase ۳ (CASP۳), and Phosphoinositide-۳-kinase regulatory subunit ۳ (PIK۳R۳) was performed using quantitative RT-PCR.Results: HCQ treatment reduced cell viability and increased apoptosis compared to both control cells and cells treated with TRE. Gene expression analysis showed a significant down-regulation of BRAF expression in cells treated with HCQ and BRAF (V۶۰۰E)-siRNA compared to siRNA-only treated cells. CASP۳ expression was significantly up-regulated in cells treated with combined HCQ and siRNA, indicating a stronger apoptotic response. PIK۳R۳ expression showed no significant change in the transfected groups.Conclusion: TRE, either alone or combined with siRNA, showed limited efficacy and may counteract the apoptotic benefits of HCQ. Conversely, HCQ, whether used alone or in combination with siRNA, enhanced apoptosis, suggesting promise as a potential treatment for melanoma.

کلیدواژه ها:

BRAF (V۶۰۰E) ، Hydroxychloroquine ، Melanoma ، siRNA ، Trehalose