摘要
目的:研究siRNA表达质粒沉默Survivin基因的效率及其对大肠癌细胞株SW480凋亡、增殖和侵袭性的影响。方法:构建Survivin基因的siRNA表达质粒(pRNAT/sur-siRNA),用Westeron-blot和半定量RT-PCR研究该表达质粒转染SW480后Survivin蛋白和mRNA表达的变化,流式细胞仪检测Survivin基因沉默后SW480细胞凋亡的变化,MTT法检测SW480细胞体外增值的变化;细胞侵袭性实验研究SW480细胞的侵袭性变化。结果:针对Survivin的siRNA的表达质粒下调大肠癌SW480细胞株Survivin蛋白和mRNA的表达水平,分别为85.0%,80.0%;pRNAT/sur-siRNA转染SW480后,SW480细胞凋亡率为16.9%;细胞增殖抑制率为37.4%,与对照组相比有显著差异(P<0.01);细胞侵袭实验示pRNAT/sur-siRNA/SW480细胞、pRNAT/SW480、SW480细胞的细胞穿透数分别为153±66、505±65、578±98个,(P<0.01)。结论:针对Survivin的siRNA可以显著降低Survivin的表达,从而抑制肿瘤细胞的增殖和侵袭,促进肿瘤细胞的凋亡。
Objective： To study the efficiency of silencing survivin gene expression level of survivin small RNA expression plasmid and its impact on the apoptosis, proliferation and invasive ability of colorectal cancer cell line SW480. Methods： The expression plasmid of survivin siRNA （pRNAT/ sur-siRNA） was constructed and transduced into SW480 cell line, was measured by the Western blot analysis and semi-quantitative reverse transcription polymerase chain reaction （RT-PCR） were used to measure the change in expression level of the survivin mRNA and protein, after transduction of the expression plasmid into the SW480. The apoptosis, proliferation, and invasive ability of SW480 cell line was evaluated by flow cytometry, MTT assay and Boyden Chamber Assay, respectively. Results： The expression plasmid （pRNAT/sur-siRNA） against survivin siRNA downregulated the survivin expression level of protein and mRNA dramatically in SW480 cell line, with a down-regulation of 85% and 80%, respectively. After transfection of the pRNAT/sur-siRNA in SW480, the apoptosis rate of pRNAT/sur- siRNA/SW480 was16.9%, proliferation of pRNAT/sur-siRNA/SW480 cell was 37.4% and there was a remarkable difference comparing with the controls （P〈0.01）. Cell invasive experiment （Boyden Chamber Assay） showed that cell penetrating membrane number of the pRNAT/sur-siRNA/SW480, pRNAT/ SW480 and the SW480 cell was 153±66, 505±65 and 578±98, respectively （P〈0.01）. Conclusion： The siRNA against survivin can significantly inhibit the expression level of the survivin, thus promote apoptosis of SW480 cell and inhibit proliferation and invasive ability of the SW480 cell.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2006年第23期1329-1333,共5页
Chinese Journal of Clinical Oncology
基金
国家自然科学基金资助(编号:30171053)
关键词
生存素
RNA
干扰
结肠癌
基因治疗
Surviving RNA interference Colon carcinoma Gene therapy
