PTEN基因联合多西环素抑制黏液表皮样癌细胞系端粒酶活性的研究

PTEN tumor suppressor gene combined with doxycycline inhibites telomerase activity in human mucoepidermoid carcinoma cell line

目的探讨外源性PTEN抑癌基因联合多西环素对人黏液表皮样癌细胞系端粒酶活性的影响。方法用脂质体将野生型PTEN基因导入黏液表皮样癌细胞系,再用不同质量浓度的多西环素处理细胞,采用MTT比色法测定细胞存活,用端粒酶重复扩增法-酶联免疫吸附(TRAP-ELISA)测定细胞端粒酶活性。结果与对照细胞比较,野生型PTEN基因明显增加癌细胞对多西环素的敏感性,增敏比为1.65～4.75倍。PTEN基因转染或多西环素诱导癌细胞端粒酶活性明显下降(P<0.05),二者联合应用对癌细胞端粒酶活性抑制更为显著(P<0.01)。结论 PTEN抑癌基因联合多西环素对人黏液表皮样癌细胞系端粒酶活性具有显著的协同抑制效应。

Objective To investigate the effect of PTEN tumor suppressor gene combined with doxycycline on telomerase activity in human mucoepidermoid carcinoma cell line.Methods The wild-type PTEN tumor suppressor gene or empty vector was introduced into mucoepidermoid carcinoma cell line in vitro,then the cancer cells were treated with doxycycline.Cancer cell survival was determined by MTT assay.Telomerase activity was determined using telomerase repeat amplification protocol-enzyme-linked immunosorbent assay（TRAP-ELISA）.Results Compared to the control cells,cancer cells transfected with the wild-type PTEN gene showed growth inhibition and increased sensitivity to doxycycyline,and the ratio of augment of drug sensitivity was 1.65-4.75.The telomerase activity in cancer cells treared with PTEN gene transfection or doxycycline alone decreased,however,telomerase activity in combined group decreased more remarkably.Conlusion PTEN gene in combination with doxycycline has significant inhibitory effect on telomerase activity in cancer cells.