siRNA沉默Livin基因促骨肉瘤MG-63细胞凋亡的实验研究
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广东省自然科学基金资助(1045101-8201004365);广州医学院博士启动基金资助(2008C21)


RNAi-mediated gene silencing of Livin enhance apoptosis of MG-63 osteosarcoma cells
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    目的:研究特异性小分子干扰RNA(siRNA)沉默骨肉瘤MG-63细胞Livin基因对细胞凋亡的影响-方法:设计针对人源Livin基因的siRNA,转染至对数生长期的人骨肉瘤MG-63细胞株;RT-PCR检测转染后骨肉瘤MG-63细胞Livin基因表达的变化,Western blot检测转染后骨肉瘤MG-63细胞Livin蛋白表达的变化,PI染色后流式细胞仪检测转染后细胞凋亡的变化-结果:Livin特异性siRNA转染骨肉瘤MG-63细胞后,Livin基因表达较空白对照和阴性对照显著减少(0.195 ± 0.019 vs 0.539 ± 0.031-0.438 ± 0.029)-Western blot检测结果显示Livin蛋白质水平较空白对照和阴性对照显著减少(0.165 ± 0.022 vs 0.632 ± 0.034-0.625 ± 0.035)-流式细胞仪检测见siRNA组骨肉瘤MG-63细胞凋亡率较对照组明显增加-结论:RNA干扰能有效沉默骨肉瘤MG-63细胞Livin基因表达,促进骨肉瘤MG-63细胞凋亡-

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    Objective: To observe the effects of silencing of Livin gene expression on the apoptosis of MG-63 osteosarcoma cells. Methods: Double stranded RNA targeting the Livin gene was chemically synthesized in vitro and transfected into MG-63 osteosarcoma cells by LipofectamineTM 2000. The transfection efficiency was observed by fluorescence confocal microscopy. Reverse transcription-polymerase chain reaction(RT-PCR) and Western blot were used to detect the expression of Livin at mRNA and protein levels. Apoptosis analysis was performed by annexin V staining. Results: The Livin mRNA expression of the Livin-siRNA transfected group was 0.195 ± 0.019,significantly lower than those of blank control(0.539 ± 0.031) and the negative control group(0.438 ± 0.029). The Livin protein expression of the Livin-siRNA transfected group was 0.165 ± 0.022,significantly lower than those of blank control(0.632 ± 0.034) and the negative control group (0.625 ± 0.035). Increased apoptotic cells were observed by flow cytometry. Conclusion: siRNA can inhibit Livin expression of MG-63 osteosarcoma cells and induce cell apoptosis. Livin might serve as a target for apoptosis-inducing therapy of osteosarcoma.

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谢楚海,吴波以,赵 玉,范子文,朱振柠. siRNA沉默Livin基因促骨肉瘤MG-63细胞凋亡的实验研究[J].南京医科大学学报(自然科学版),2011,(11):1610-1613

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  • 收稿日期:2011-07-01
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