Study on signal transduction in apoptosis of human melanoma cells A375 induced by apoptin

ObjectiveTo study whether apoptin gene induces apoptosis in human melanoma cells A 375 via the activation caspase-8 and caspase-3.MethodsThe human melanoma cells A 375 were transiently transfected with recombinant pcDNAA 3 plasmid, which contained apoptin gene,and the apoptosis was measured by DNA agarose electrophoresis,RT-PCR and flow cytometry.Caspase-8 and caspase-3 relative activity was analyzed by colorimetric assay at various time.ResultsThe apoptin gene could induce apoptosis of human melanoma cells A 375 in vitro and the mRNA of apoptin gene could be detected by RT-PCR.The activity of caspase-3 began to rise in A 375 cells after transfected with apoptin gene,but that of caspase-8 did not change.ConclusionApoptin gene can induce apoptosis in human melanoma cells A 375 via active caspase-3.