Anti-tumor Effect Of Recombinant Lentivirus GV493-hTERT-shRNA On Malignant Melanoma

Objective:The aim of this study is to investigate the anti-tumor effect of hTERT knockdown of human malignant melanoma and its potential clinical value.Methods:After constructing lentivirus vector to interfere with hTERT,cells were cultured in groups and divided into blank control group:The A375 cells without any intervention;Negative control group:The A375 cells were infected with empty vector of virus;GV493-hTERT-shRNA group:Lentiviral vector of hTERT knockdown infected A375 cells.In vitro,RT-qPCR and Western-blot assay were performed to detect the expression of target gene.Then we performed CCK-8 assay to detect cell proliferation activity,transwell assay and scratch assay to detect cell invasion and migration ability,and flow cytometry to detect cell apoptosis and cell cycle.Simulated melanoma growth via a xenografts mouse model in vivo after employing hTERT knockdown strategy.Results:The expression of target gene and the proliferation,migration and invasion ability of GV493-hTERT-shRNA group were lower than that of other two groups,the difference has statistic significance(P<0.05).But the apoptosis of GV493-hTERT-shRNA group was higher than that of other two groups,G0/G1 phase arrest occurred,and the difference has statistic significance(P<0.05).The tumor growth of GV493-hTERT-shRNA group was slower than that of the other two groups(P<0.05).Conclusion:hTERT knockdown strategy was successfully employed by lentiviral vector.The ability of melanoma cell about proliferation,invasion and migration were weakened and apoptosis was increased after interfering the expression of hTERT,which effectively inhibited tumor growth and metastasis in xenograft mouse.It is suggested that hTERT plays an important role in the occurrence and development of melanoma,and hTERT is expected to become a new potential target or related biomarker for the treatment of melanoma.