端粒酶活性在哺乳动物细胞中的表达

Expression of the Activity of Telomerase in Mammal Cells

为研究端粒酶活性在哺乳动物细胞中的激活 ,构建了端粒酶催化亚基 (human telomerase reverse transcriptase,h TERT)和端粒酶 RNA组分 (h TR)基因的真核表达载体 ,分别在无端粒酶活性的 DK细胞株和人肝癌细胞中进行表达 ,以 TRAP-银染和 TRAP- EL ISA定量方法测定转染细胞。结果显示 ,在人肝癌细胞株中仅 h TERT表达即可使端粒酶活性显著增加 ,而 DK细胞中 h TERT必须和 h TR同时转染才有端粒酶活性表达。这一结果表明 ,端粒酶活性表达主要和 h TERT/ h TR有关 ,采用人的端粒酶 h TERT/ h TR在其他哺乳动物细胞中表达端粒酶活性是可行的。

To study the activation of telomerase in mammal cells,we constructed the eukaryotic expression vector of telomerase catalytic s ubunit and the RNA component gene,and transfected into negative DK cell and huma n liver cancer cells.The telomerase activity was assayed by TRAP-ELISA and silv er stain.The results showed that telomerase was increased significantly in human liver cell when transfected with hTERT only,and the activation in DK cell must transfected with both hTERT and hTR indicating the telomerase acitivity is assoc iated with these two genes,and activation of telomerase in other animal cells wi th hTERT/hTR is possible.