The Expression Of Caspase3 In Leukemia K562 Cell Through Silence UVRAG Induced Diallyl Disulfide

Objective:Research UVRAG gene mediated autophagy in DADS induced the change of leukemia K562 cells apoptosis,Preliminarily study on induced by DADS the relationship between apoptosis and autophagy in K562 cells.Methods:K562 cells were cultured in K562 cell culture systems.K562 cells were treated with DADS(20mg/L,40mg/L,80mg/L) for 12 hours and extracts cell protein.the protein imprinting technology(westernbloting) is used to detect the expression of apoptosis-related genes of caspase-3;successfully constructed and selected the most effective gene sequence fragments of siRNA about the gene of UVRAG tranfect to the K562 cells using lipofectamine TM2000-liposomal. The interference effection is observed by the QT-PCR detecting the expression levels of UVRAG mRNA after 24 hours transfection, then using 40mg/LDADS effects the group of transfection for 12 hours. Western blotting is employed to measure the expression of caspase3 protein that may be induced K562 cell apoptosis.Results:The different concentrations of DADS(20mg/L、40mg/L、80mg/L) treat with the K562 cells for 12 hours,The Western Blotting result showed The caspase3 protein expression levels of the K562 cells of treatment groups is raised than the control group(P <0.05). compared with the blank group,the UVRAG mRNA expression levels were lower,the product of mRNA decreased more obviously treated with DADS,40mg/L after 12 hours, it shows the silence of UVRAG successfully. Treat the interference successful of K562 cells with DADS,40mg/L for 12 hours,the result show the caspase3 protein expression levels of the K562 cells of treatment groups is reduced than the control group.(P <0.05)Conclusions: Autophagy induced by DADS in K562 cells depends on the expression level of inherent UVRAG gene. Inhibition of UVRAG mediated autophagy may also inhibit DADS induced apoptosis of K562 cells.