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

Abstract

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 green fluorescent 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) (3.0 μg/mL) for the same time, cell survival rate and IC₅₀ was detected with MTT, cell apoptosis was detected with fluorescence-activated cell sorting (FACS), and the nuclear alteration was observed by TdT-mediated deoxyuridine triphosphate nick end labeling (TUNEL). In addition, caspase-3 activity was detected by using colorimetric method. Cell viability was decreased significantly in GBC-SD/survivin cells, and survivin expression was decreased significantly (mRNA and protein of survivin were decreased by 74.7% and 71.5%, respectively). After treatment with DDP, cell survival rate and IC₅₀ was decreased significantly (2.03±0.24 μg/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 at first 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.

Keywords

survivin / shRNA / chemotherapy / sensibility / GBC-SD