科研之友
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摘要
Survivin has become an attractive anticancer therapeutic target due to its important role in tumor cell viability and its selective expression in tumor cells. In the present study, we constructed a recombinant siRNA plasmid vector against survivin and stably transfected it into HepG2 and SMMC-7721 hepatocellular carcinoma cells in vitro. Semi-quantitative RT-PCR and western blotting were used to determine the expression of survivin mRNA and protein, respectively. Tumor cell proliferation was assessed by trypan blue exclusion. We evaluated the change in caspase-3 activity, and the rate of cell apoptosis and the cell cycle distribution were analyzed by flow cytometry. Assessment of chemosensitivity was carried out by MTT assay. The results showed that transfection of survivin siRNA caused a significant inhibition of survivin mRNA and protein expression which was associated with cell growth inhibition, specific G0/G1 phase arrest, increased caspase-3 activity and enhanced chemosensitivity to cisplatin in both HCC cell lines. We suggest that suppression of survivin expression by RNAi attenuates the malignant phenotype of hepatocellular carcinoma cells, and may provide a novel approach for anticancer gene therapy.
- 出版日期2013-3
- 单位苏州大学
