2010•Zhonghua gan-dan waike zazhiRequires access
Effect of survivin shRNA on chemotherapy resistance in GBC-SD cells
Shen Han-bin, Yaohui Wu, Long Hao-cheng, Zheng Qichang, Gong Jianping
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Abstract
Objective To investingate the effect of survivin shRNA on chemotherapy resistance in GBC-SD cells. Methods They were divided into three groups of GBC-SD, GBC-SD/EGFP and GBC-SD/survivin. MTT assay was used to detect cell viability in the 3groups. mRNA and protein of survivin were tested by RT-PCR and Western blot. Then the cells were treated with proper construction DDP. The cell survival rate and IC_(50) were determined with MTT, cell apoptosis detected by FACS and the alteration of nucleus observed by TUNEL. Meanwhile, caspase-3 activity was determined using the colorimetric method. Results Cell viability was reduced remarkably in GBC-SD/survivin and survivin expression was decreased obviously. After being treated with DDP, cell survival rate and IC_(50) were decreased obviously in GBC-SD/survivin, apoptotic rate elevated remarkably compared with other groups. There were brownly nucleuses in three groups. Caspase-3 activity increased first and then decreased, but it exceeded in GBC-SD/survivin than that in other two groups. Conclusion The survivin shRNA can down-regulate the expression of survivin in GBC-SD cells remarkably and improve the sensibility to chemotherapy.
Key words:
Chemotherapy; survivin; shRNA; GBC-SD