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Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front. Med.    2009, Vol. 3 Issue (4) : 485-490     DOI: 10.1007/s11684-009-0089-3
Research articles |
Effect of pEGFP-survivin on GBC-SD cell growth and chemotherapy sensitivity
Hanbin SHEN MD1,Bin ZHANG MBA1,Song ZHAO BM1,Qichang ZHENG MD2,Jianping GONG MD3,Xiaotang CAI BM4,
1.Department of General Surgery, The Fifth Hospital of Wuhan, Wuhan 430050, China; 2.Department of Hepatobiliary Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China; 3.Department of Intestines and Stomach Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China; 4.Department of Surgery, The 477th Hospital of People''s Liberation Army, China;
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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      
Issue Date: 05 December 2009
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