hTERT基因反义cDNA转染对卵巢癌细胞系SKOV3生物学行为的影响

目的: 研究反义hTERT基因对卵巢癌细胞系SKOV3生物学行为的影响.方法: 构建反义hTERT基因的真核表达载体,采用LIPOFCTAMINETM Reagent转染卵巢癌细胞系SKOV3,通过RT-PCR、Trap-ELASA、生长曲线、裸鼠体内致瘤能力等方法比较转染前后细胞hTERT基因表达、端粒酶活性、细胞生长、致瘤性等方面的变化.结果:转化细胞系细胞hTERT基因的表达受到抑制,端粒酶活性下调,肿瘤细胞的增殖与生长速度明显降低,细胞的克隆形成能力及裸鼠体内的成瘤能力下降,肿瘤的恶性表型部分逆转.结论:应用反义技术下调hTERT基因表达可有效降低端粒酶活性并部分逆转卵巢癌细胞的恶性表型,hTERT基因可望成为卵巢癌基因治疗的新靶点.

Biologic effects of antisense hTERT expressing vector on human ovarian cancer cell line SKOV3

Objective: To study the effects of antisense hTERT expressing vector on biologic behavior of human ovarian cancer cell line SKOV3.Methods: Recombinant eukaryotic expression vector pcDNA3 AS hTERT with full length hTERT antisense cDNA and vector pcDNA3 were introduced into SKOV3 which highly expressed hTERT by lipofectamine mediated gene transfection. hTERT gene expression,telomerase activity,rates of cell growth and tumorogenesis in nude mice were compared before and after SKOV3 being transfected. Results: The transformant cells showed retardation of cell proliferation,growth and partial reversion of the malignant phenotype,including decrease of hTERT gene expression ?telomerase activity and rates of cell growth,and also the ability of soft agar colony formation & tumorogenesis in nude mice. Conclusion: Down regulation of hTERT can effectively decrease telomerase activity and partially reverse malignant phenotype in SKOV3.hTERT may be a new and potent target of gene therapy in ovarian cancer.