Experimental Study Of The Interfering Effect On EJ-28 Cell Line Induced By Sirna Targeting Survivin

Objective To explore proliferation and apoptosis of EJ-28 cells induced by different siRNAs targeting survivin.Methods 3 siRNAs targeting survivin and 1 fluorescence-labeled siRNA as a nagative control were chemically synthesized . Transfetion rate was observed by fluorescence microscope after transfecting the fluorescence-labeled siRNA into EJ-28 cells with lipofectamine- RNAi-Mate. The effect of siRNA suppressing the proliferation of EJ-28 cell was detected by MTT assaay at 24h,48h,72h after transfection;Apoptotic rate was detected by Annexin V-FITC-FCM assay at 48h after transfection.The most efficient siRNA picked out by former research was transfected into EJ-28 cells with lipofectamine- RNAi-Mate.At 48h after transfection with the siRNA,the ultrastructure of EJ-28 cells was studied by transmission electron microscope;the cell cycle of EJ-28 cells was detected by Flow Cytometry;The change of survivin protein expression was detected by immunofluroescence technique.Results1.All the sequence-specific siRNAs targeting to survivin can efficiently suppress the proliferation of EJ-28 cell,moreover siRNA-164 can most significantly suppress the proliferation comparing with siRNA167 and siRNA389;The highest inhibition rate was (41.32±2.54)% at 48h after the start of transfection;the highest apoptotic rate induced by siRNA164 was (13.23±0.25)% at 48h after the start of transfection.2. There were typical apoptotic features of morphology in EJ-28 cells transfected with siRNA164,including reduction in volume, nuclear chromatin condensation along nuclear periphery and apoptotic bodies found under transmission electron microscopy.3.Comparison with control group , the cell cycle of EJ-28 cells transfected with siRNA164 was mainly arrested at G2/M phase,that was significant difference( P < 0. 05).4.The mean fluorescence intensity of siRNA group (31.04±0.95)was significantly lower than that of negative control group(55.01±1.27).The protein level of survivin was decreased significantly ( P < 0. 05), and the inhibition rate was (43.58±1.05)%Conclusions 1.Blocking the expression of survivin with RNA interfering technology can significantly suppress proliferation of EJ-28 cells and induce apoptosis to a certain degree,moreover siRNA-164 is the most efficient one.2.SiRNA targeting survivin can significantly suppress the proliferation of EJ-28 cell probably by changing cell cycle and blocking the expression of survivin at protein level.3. RNAi targeting survivin has a potential value in gene therapy of bladder cancer.