Effect of B-RAF⁶⁰⁰ activating gene mutations on the ability of melanoma cells to autophagy

Background. We investigate the interaction of the B-RAFgene mutations status and levels of autophagy induction in melanoma cell lines in vitro. Methods. In this report 12 human melanoma cell lines were investigated. B-RAF gene mutations were determined by PCR, expression of Beclin1 mRNA-by RT-PCR. Analysis of the LC3B protein expression and accumulation, specifically associated with the membranes of autophagosomes, was performed by immunocytochemical assay. Results. Among melanoma cell lines 7 had activating mutations in B-RAF gene. Basal expression level of Beclin1 mRNA was identified in all melanoma cell lines. The basal expression ranged from 0,15 to 1,16 r.u. Cell lines starvation resulted in increased expression of mRNA Beclin1 level in cells B-RAF^wt, but B-RAF^V600 cell lines, it remained almost unchanged. Autophagy induction by temozolomide has shown a significant increase in the amount of autophagosome punctuate in B-RAf and B-RAF⁶⁰⁰ melanoma cells. Conclusion. B-RAF mutations may affect the melanoma autophagy level in vitro. Investigation of the mechanisms of this process will enable the development of new predictive factors of drug resistance in melanoma and identify the original target for anticancer therapy.

аутофагия, меланома, мутации в гене B-RAF, химиорезистентность, autophagy, melanoma, B-RAF gene mutations, chemoresistance