Effect of pEGFP-survivin on GBC-SD cell growth and chemotherapy sensitivity

This paper is aimed to investigate the effect of survivin shRNA on chemotherapy resistance in human gallbladder carcinoma GBC-SD cells.The viability of human gallbladder carcinoma GBC-SD,GBC-SD/enhanced greenfluorescent protein(EGFP),and GBC-SD/survivin cells was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay,and mRNA and protein of survivin were tested by reverse transcription-polymerase chain reaction(RT-PCR)and Western blot.After the cells were treated with cisplatin(DDP)(3.0μg/mL)for the same time,cell survival rate and IC50 was detected with MTT,cell apoptosis was detected withfluorescence-activated cell sorting(FACS),and the nuclear alteration was observed by TdT-mediated deox-yuridine triphosphate nick end labeling(TUNEL).In addition,caspase-3 activity was detected by using colorimetric method.Cell viability was decreased sig-nificantly in GBC-SD/survivin cells,and survivin expres-sion was decreased significantly(mRNA and protein of survivin were decreased by 74.7%and 71.5%,respec-tively).After treatment with DDP,cell survival rate and IC50 was decreased significantly(2.03�0.24μg/mL)in GBC-SD/survivin cells,while apoptotic rate(84.3%)was elevated significantly as compared with the other two groups.There were brown apoptotic nuclei in all the cells.Caspase-3 activity in all the cells was increased atfirst and then decreased,but the caspase-3 activity in GBC-SD/survivin cells was significantly higher than the other two groups.The survivin shRNA could down-regulate the expression of survivin in GBC-SD cells significantly and improve the sensibility to chemotherapy.