Effects Of Lentiviral Vector RNAi Silencing IGHG1 Gene On The Cell Function Of Bladder Cancer EJ Cells

【Objective】 To investigate the expression of Ig G in bladder cancer cells and the effects of IGHG1 targeted gene silencing lentiviral vector on cell migration,proliferation and apoptosis of human bladder cancer EJ cells.【Methods】 This research investigate the expression of Ig G in bladder cancer cells by WB q PCR and IF.bladder cancer cell line EJ had been transfected with LV-IGHG1-RNAi lentiviral vector to use as experimental group(sh IGHG1-EJ cells).Negative scrambled lentiviral vector was used to transfecting bladder cancer cell line EJ,which treated as negative control group(NC-EJ cells).EJ cells were used to be blank control group(EJ cells).This research verified the expression of Ig G γ by Western blot(WB)and Immunofluorescence(IF);IGHG1 m RNA expression were detected by real-time Quantitative PCR(q PCR).Cell Counting Kit 8(CCK-8)were used to detect the proliferation of these three cells.Flow cytometry method(FCM)were used to detect the apoptosis of cells;WB were used to detect the apoptins caspase-3 and cleaved caspase-3,which means the apoptosis of cells,too.Wound healing assay were used to investigate the change of sh IGHG1-EJ cell’s migration.【Results】 The expression of IGHG1 m RNA and Ig G γ of T24 and EJ cells were more than SV-HUC-1cell(P<0.001).The transfecting rate of sh IGHG1-EJ cell was 76.667%±0.042%,and the rate of NC-EJ cell was 75.333%±0.055%.The expression of IGHG1 m RNA and Ig G γ of sh IGHG1-EJ group were obviously lesser than control groups(P<0.01).It shows that IGHG1 gene silencing were successful.CCK-8 revealed that IGHG1 gene silencing significantly decreases the proliferation of EJ cells(P<0.01).FCM and WB found that IGHG1 gene silencing can promotes the apoptosis of EJ cells(P<0.05).Wound healing assay shows that Cell migration inhibited by IGHG1 gene silencing(P<0.01).These assay of NC-EJ cells had no meaningful different from EJ cells.【Conclusions】 Bladder cancer cells and urothelial cell can expression Ig G protein and IGHG1 m RNA.IGHG1 gene silencing can observably inhibits the cell migration and proliferation of human bladder cancer EJ cell;It also can promote cell apoptosis.This may serve as a new target for bladder cancer targeted therapy.