Effects Of RNA Interference In Apoptosis Inhibitor Livin On Bladder Cancer Cell Apoptosis And Cell Cycle

Inhibitor of Apoptosis Protein (IAPs) is a class of anti-apoptotic factor in vivo. Livin is a newly discovered member of IAPs, almost no-expression in normal adult tissues, but is highly expressed in many cancers, and is considered have important links on tumors’poor prognosis and drug resistance. Studies have shown that not only has anti-apoptotic effect, the RING finger domain in its molecular structure given Livin ubiquitin-conjugating enzyme-like activity, makes it an important role in the regulation of cell cycle. Research of the expression of Livin gene in bladder cancer and the impact of Livin gene in bladder cancer cell apoptosis insufficiency and chemotherapy resistance is still lacking. The impact of Livin expression on bladder cancer apoptosis and cell cycle needs further study.Objective:(1) screening out an efficient and specific siRNA for Livin gene.(2) Interfering Livin gene expression by RNAi in bladder cancer cells, observing the impact of Livin gene expression decrease on bladder cancer cell apoptosis and cell cycle.Methods: (1) by using siRNA design tools such as siRNA design software and siRNA database design Livin specific siRNA. Filter out an efficient and specific siRNA by RT-PCR and Western blot.(2) By Bivariate flow cytometry, detecting the impact of siRNA interference induced Livin gene expression decrease on human bladder cancer T24cells apoptosis.(3) By flow cytometry, detecting the impact of siRNA interference induced Livin gene expression decrease on human bladder cancer T24cells cell cycle.Results:1. Livin gene specific siRNA screeningUsing siRNA design software and siRNA databases, we obtained three kinds of siRNA, through RT-PCR and Western blot test, all three siRNAs could partly reduce the expression of Livin gene, in which one kind can significantly reduce the expression of Livin gene.2. The effects of siRNA interference induced Livin gene expression decrease on human bladder cancer T24cells apoptosisResults of bivariate flow cytometry showed that Livin gene expression decrease resulting in an obviously increase in the apoptosis of T24cells.3. The effects of siRNA interference induced Livin gene expression decrease on human bladder cancer T24cells’cell cycleResult of flow cytometry showed that Livin gene expression decrease resulting in T24cell cycle arrested at G2/M phase. Conclusion:1. Decrease the expression of Livin gene in T24bladder cancer cells, could effectively inhibit the proliferation of T24cells.2. Decrease the expression of Livin gene in T24bladder cancer cells, could obviously increase the spontaneous apoptosis of T24cells.