The effect of Survivin shRNA on chemotherapy resistance of GBC-SD cells

Objective To investigate the effect of Survivin shRNA on chemotherapy resistance of GBC-SD cells. Methods They were divided into three group which are GBC-SD, GBC-SD/EGFP and GBC-SD/survivin. MTT assay was used to detect cells viability in three groups,and mRNA and protein of survivin were tested by RT-PCR and Western blot. Then cells were treated with proper construction DDP (3.0 mg/L) for the same time,cell survival rate and IC_(50) was detected with MTT respectively,cells apoptosis was detected through FACS.and the alteration of nucleus was observed by TUNEL. In addition, caspase-3 activity was detected by using colorimetric method. Results Cell viability was reduced remarkably in GBC-SD/survivin and survivin expression was decreased obviously (74.7% ,71.5%). After treated with DDP,cell survival rate and IC_(50) was decreased obviously [(2. 03 ±0. 24) mg/L] in GBC-SD/ survivin, apoptosis rate (84.3% ) were elevated remarkably compared with other group. There were brownly nucleuses in three groups. Caspase-3 activity all ascend first and then descent,but it exceeded in GBCSD/survivin than that in other two groups. Conclusion The survivin shRNA could down-regulate the expression of survivin in GBC-SD cells remarkably and improve the sensibility to chemotherapy. The alteration of caspase-3 activity followed with survivin contrarily. Key words: Carcinoma of gallbladder; Survivin; shRNA; Chemotherapy