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目的 :观察Survivin反义核酸对人卵巢癌细胞系SKOV3的生长及Survivin基因表达的影响。方法 :应用基因重组技术 ,将在真核细胞中能稳定表达反义Survivin的pcDNA3 SVVas经脂质体LipofectamineTM2 0 0 0 介导转染人卵巢癌细胞株SKOV3,经G4 1 8筛选得到抗性克隆 (SKOV3/SVVas) ,同时以空载体pcDNA3转染SKOV3作为对照(SKOV3/neo) ;绘制细胞生长曲线 ;采用免疫组化、RT PCR和流式细胞仪检测SKOV3、SKOV3/neo及SKOV3/SVVas细胞的内源性SurvivinmRNA及蛋白的表达和凋亡细胞。结果 :与SKOV3及SKOV3/neo细胞比较 ,SKOV3/SVVas细胞增殖和代谢能力均明显降低 ,Survivin蛋白阳性表达率及mRNA相对表达量明显降低 (P <0 .0 1 ) ,细胞凋亡率显著增加 (P <0 .0 1 )。结论 :稳定表达的反义Survivin能够有效抑制SKOV3细胞内源性Survivin蛋白的表达 ,诱导SKOV3细胞凋亡。
Objective: To observe the effect of Survivin antisense oligodeoxynucleotide on the growth of human ovarian cancer cell line SKOV3 and the expression of Survivin gene. METHODS: pcDNA3 SVVas stably expressing antisense Survivin in eukaryotic cells were transfected into human ovarian cancer cell line SKOV3 by LipofectamineTM 2000, and the resistant clones were screened by G418 (SKOV3 / SVVas) were transfected with empty vector pcDNA3 as control (SKOV3 / neo); cell growth curve was drawn; SKOV3, SKOV3 / neo and SKOV3 / SVVas cells were detected by immunohistochemistry, RT PCR and flow cytometry Endogenous Survivin mRNA and Protein Expression and Apoptotic Cells. RESULTS: Compared with SKOV3 and SKOV3 / neo cells, SKOV3 / SKOV3 / neo cells significantly reduced the proliferation and metabolism, the expression of Survivin protein and the relative expression of mRNA were significantly decreased (P <0.01), and the apoptosis rate was significantly increased (P <0. 01). Conclusion: The stable expression of antisense Survivin can effectively inhibit the expression of endogenous Survivin protein in SKOV3 cells and induce the apoptosis of SKOV3 cells.