摘要
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 vs0.539±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 vs0.632±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。
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.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2011年第11期1610-1613,1631,共5页
Journal of Nanjing Medical University(Natural Sciences)
基金
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)
