Investigating Therapeutic Efficacy of Dacarbazine and Temozolomide, Alone and in Combination with BRAF(V600E) siRNA in A375 Human Melanoma Cell Line

Melanoma is one of the variable nature and deadliest form of skin cancers and the mutations play a major role in its development. It has faced many challenges in treatment and has led to the orientation of individual therapies to combined therapy. the present study is aimed at determining the combination therapeutic effects of Dacarbazine, Temozolomide with siRNA BRAF (V600E) in the A375 as treatments for melanoma. The A375 cell line was treated with different concentrations of drugs for 24-72 hours.The IC50 and viability%, apoptosis%, and expression level of BRAF , CASP3 , and PIK3R3 genes were examined respectively by MTT, flow cytometry and qRT-PCR. Results : The cell viability % in the chemotherapy groups and siRNA with drugs groups has a substantial decrease, no significant difference was observed in the overall comparison of the two groups. The percentage of apoptosis in both chemotherapy-treated groups and siRNA + drugs indicated a significant difference compared to the control. The expression level of BRAF gene had a notable decrease in expression in the BRAF (V600E) siRNA-transfected cells with drugs compared to the chemotherapy groups. The most effective enhancement was noted in the siRNA+DTIC+TMZ group (22.43±0.9884). Our exploration extended to the mean expression levels of the PIK3R3 gene in the combined siRNA+chemotherapy groups, revealing a notable reduction. Conclusions : these results would seem to implicate that the combination of siRNA plus chemotherapy in the A375 cell line is encouraging for the development of effective therapeutic strategies and should be explored and validated more to improve melanoma treatment.