Effects of RNA Interference to Selenoprotein V Gene on Cytobiological Behaviors in Human Malignant M

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  To investigate the effects of RNA interference to selenoprotein V (SELV) gene on cytobiological behaviors in human malignant melanoma A375 cells.the recombinant lentivirus vector containing SELV-shRNA was constructed and transfected into A375 cells.The cells constantly expressing the shRNA was selected after incubation with puromycin.The expressions of SELV mRNA and protein were detected by quantitative real-time reverse transcription PCR (qRT-PCR) and Western blottingrespectively.The proliferation of transfected A375 cells was assayed by MTT test.Cell cycle and apoptotic rate were analyzed by flow cytometry.The expressions of ankyrin repeat and SOCS box containing 9 (ASB9) and O-acetylglucosamine transferase (OGT),two SELV functionally related factors,were determined by qRT-PCR and ELISA,respectively.Compared with the control,significantly decreased expressions of SELV mRNA and protein (P<0.05) were found in transfected A375 cells,while the apoptotic rate was increased significantly (P<0.05).However,there was no significant difference in cell cycle and proliferation between the two groups.After the SELV gene was suppressed,the mRNA and protein expressions of ASB9 and OGT were increased significantly (P<0.05).In A375 cells,down-expression of SELV gene led to increased apoptotic rate,which was accompanied by elevated expression of ASB9 and OGT genes.
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