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Survivin gene-targeted siRNA inducing apoptosis of human osteosarcoma cell line U-2OS
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Abstract:
To investigate the proliferation and apoptosis of human osteosarcoma cell line U-2OS after sur-vivin gene was knocked down by siRNA synthesized in vitro. Methods: U-2OS cells were divided into 3 groups: Group A (blank control) , group B ( transfected with non-specific siRNA) and group C( transfected with survivin-specific siRNA). The morphological changes of U-2OS cells were observed with fluorescent microscope. The expression of survivin mRNA and protein were detected by reverse transcription polymerase chain reaction ( RT-PCR) and Western blotting. The anti-proliferative effects were assessed by MTT and the rate of apoptosis was examined by flow cytometry in 3 groups. The results of the 3 groups were analyzed and compared. Results: Survivin specific siRNA significantly down-regulated mRNA and protein expression of survivin. Expression of survivin protein in group C was significantly lower than those in group A and B( P <0. 01 ) ; Group C also showed a slower proliferation than the other 2 groups ( P <0. 01 ). The apoptosis rate in group C was significantly higher than those in other 2 groups ( P <0.01 ). Conclusion: siRNA can efficiently suppress survivin gene expression, inhibit proliferation and induce apoptosis in osteosarcoma cell line U-2OS.
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