Effects Of MicroRNA-100 On The Proliferation Of Bladder Cancer T24cells And The Regulation Of VEGF And Surviving Protein

Objective: microRNA-100(mi RNA-100) for bladder cancer T24 cells of vascular endothelial growth factor(vascular endothelial growth factor, VEGF), and regulation of survivin protein and m RNA expression determined by experiment, explore the mi R-100 on bladder cancer T24 cell proliferation and apoptosis.Methods: synthetic mi R-100 double-stranded complementary DNA fragments corresponding insertion p Silencer TM 3.1-H1 neo vector expression vector after sequencing as micro RNA expression plasmid; the use of transient transfection method Lipofeetamine TM2000(liposomes 2000) mi R-100 expression plasmid transfected into bladder cancer T24 cells were cultured for 24 hours after selecting successfully transfected cell lines continue higher survival training. By RT-PCR, and Western blot,respectively, after 48 hours of incubation to detect the m RNA and protein expression of VEGF and survivin cell lines, with the tetrazolium blue(MTT) to detect bladder cancer T24 cells 24 h law, 36 h, 48 h and 60 h proliferation changes. Then using Annexin V / PI double staining flow cytometry to detect bladder cancer cell apoptosis in T24. Statistically analyzed with each set of experimental data results.The result:1. Western blot showed that mi R-100 can inhibit VEGF(6.20±1.13)%, P <0.05. And survivin protein expression(5.60±1.02)%, P<0.05.2.RT-PCR analysis showed VEGF(9.85±3.20)%, P <0.05. And survivin m RNA levels decreased(8.22±3.21)%, P<0.05.3.MTT display transfected cell growth rate than the non-transfected cells was significantly slowed down, continued for 60 hours after the inhibition rate(11.54±4.07)%, P<0.05.4. Flow cytometry showed: T24 cell apoptosis after transfection of mi R-100 in thecontrol group than in the control group and increased(46.35 ± 2.14%), P <0.05. Cell cycle G0/G1 phase cell ratio increased.In conclusion:1.mi R-100 inhibits the proliferation of T24 bladder cancer cells, suggesting that mi R-100 RNA inhibited the expression of VEGF and survivin gene.2.mi R-100 can promote T24, apoptosis may be related to cell cycle arrest.