The Effect Of The SiRNA Lentivirus Vector Targeting Survivin On Human Malignant Melanoma Cell Line A375

Background Malignant melanoma is a highly malignant skin cancer, with early onset age and high transfer rate, and it is insensitive to chemotherapeutic drug.Along with the development of modern medicine, The gene therapy of Malignant melanoma become the present study hotspot.Survivin is a new member of the inhibitor apoptosis protein(inhibitor of apoptosis, IAP)family, which is the most powerful apoptosis inhibitor in currently found. It is specifically expressed in the tumor tissue,but not expressed in the normal tissue.It has dual role, inhibiting apoptosis and regulating cell mitosis.RNA interference surppresses the expression of target gene using small double-stranded RNA, promotes the degradation of target gene mRNA efficiently and specifically,thus reaches the gene silencing. So it is widely used in the study of gene therapy.Objective To observe the effect of the RNA interference (RNAi) targeting survivin on human malignant melanoma cell line A375。Methods To design and construct the shRNA lentivirus vector targeting survivin, transfect it into human malignant melanoma cell line A375, infection efficiency was observed by microscope. protein expression of survivin was detected by western blot, and mRNA expression of survivin was detected by real-time PCR.Results The survivin-siRNA lentivirus vector was designed and constructed successfully,72h after the transfection, the infection efficiency was higher than80%,the expression of survivin mRNA and protein were significantly lower in the survivin-siRNA-LV group than in the control group (P<0.05), inhibition ratio was60%、87.38%,the expression of survivin mRNA and protein were significantly lower in the survivin-siRNA-LV group than in the NC-GFP-LV group (P<0.05), inhibition ratio was59.75%、86.72%,the expression of survivin mRNA and protein were no difference between the NC-GFP-LV group and the control group (p>0.05)Conclusion the siRNA targeting survivin can down-regulate the expression of survivin gene in A375cells。...