1249 Research of hiperfect-mediated RNA interference targeting hTERT on the biological behavior of the human malignant melanoma cell line A375

Objective: To Research of hiperfect-mediated RNA interference targeting human telomerase reverse transcriptase gene (hTERT) on the biological behavior malignant melanoma cell line A375.Methods: Specific siRNAs with green fluorescent labeling different concentrations were transfected into A375 cells by using transfection reagent (Hiperfect) at volumes . The efficiency was determined fluorescen microscop, and concentration siRNA volume Hipetfect selected. Four hTERT-siRNAs constructed separately most optimal condition, extraction total mRNA after 24-hour treatment. Real-time fluorescence-based quantitative PCR conducted to select specific hTERT-siRNA, MTT assay evaluate proliferation. RT-qPCR western experiments used detect hTERT protein expression levels in each group before interference. Cell scratch transwell migration invasion ability group. Results: 5nmol/L 3μl Hiperfect significantly higher than other groups(all P＜0.05).The hTERT-siRNA-203 showed lowest (all P＜0.05),and best. proliferation inhibition rates 24, 48 72 hours from groups P＜0.01).Using can down-regulate (P <0.05). results test that also decreased Conclusion: Hiperfect-mediated affect A375.