【摘要】 目的 研究反义hTERT基因体外诱导肺癌细胞凋亡。 方法 体外通过SuperFect(QIAGEN)转染正、反义hTERT真核表达载体至人肺腺癌细胞株GLC-82，再经过G418及PCR筛选鉴定获得分别转入了正、反义hTERT载体的抗性克隆细胞GLC-82-s和GLC-82-as。然后采用MTT法、双层软琼脂克隆形成实验、流式细胞术观察和分析了反义hTERT对肺癌细胞体外生长增殖活力的影响及是否能诱导瘤细胞的凋亡;同时我们还运用实时荧光定量RT-PCR技术及TRAP-银染法对各组细胞内源性hTERT mRNA的表达情况及端粒酶的活性进行了检测。 结果 当各组细胞传至第25代后，与GLC-82、GLC-82-s比较，GLC-82-as细胞的生长速度和集落形成能力明显地减慢和降低，同时伴有凋亡率的显著增加;与空白对照、正义hTERT组相比，反义hTERT能显著地降低GLC-82细胞内源性hTERT mRNA的表达(P<0.01)和下调端粒酶活性。 结论 反义hTERT体外确实能明显地诱导肺癌细胞的凋亡及抑制瘤细胞的生长增殖能力。

    【关键词】 反义hTERT;肺癌;凋亡;细胞增殖;实时荧光定量RT-PCR

    A study on cell apoptosis of lung cancer induced by antisense hTERT in vitro

    YIN Wei-hua，MAYa，LI Ruo-xin，et al.

    Department of Pathology，Peking University Shenzhen Hospital，Shen-zhen518036，China

    【Abstract】 Objective To study the cell apoptosis of lung cancer induced by antisense hTERT in vitro.Methods The sense and antisense hTERT eukaryotic expression vectors were transfected into lung cancer cell line GLC-82using the SuperFect transfection reagent(Qiagen)according to the manufacturer's instructions，then the GLC-82-s and GLC-82-as of G418-resistant colonies were obtained with G418and identified for the presence of hTERT insert by PCR with T7and pcDNA3.1/BGH reverse primers.After that ......