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目的上调甲状腺鳞癌细胞(SW579)的人端粒酶催化亚单位(hTERT)表达,观察细胞增殖情况,检测端粒酶活性变化。方法应用酶切方法获得hTERT序列全长3 453 bp的cDNA片段,克隆入荧光蛋白质粒(pEGFP-N1),构建真核表达载体pEGFP-hTERT,转染SW579细胞中,观察细胞生长状态的变化,检测端粒酶活性。结果稳定转染pEGFP-hTERT的SW579细胞的增殖率为148%,比非转染组增加48%,比空质粒组增加45%,差异均有统计学意义(P<0.01);重组质粒转染组G1、S、G2/M期细胞比例分别为68.05%,27.66%和10.45%,与非转染组和空质粒组比较,差异均有统计学意义(P<0.01,P<0.05);重组质粒转染组增殖指数(PI)为38.11,明显高于空质粒组的14.22和非转染组的13.60,其端粒酶活性检测梯状条带增加,活性增强。结论上调人端粒酶表达,能够促进SW579细胞增殖。
Objective To investigate the expression of telomerase catalytic subunit (hTERT) in thyroid squamous cell carcinoma (SW579) cells and observe the cell proliferation and detect the change of telomerase activity. Methods A cDNA fragment of 3 453 bp in length of hTERT gene was obtained by enzyme digestion and cloned into pEGFP-N1 plasmid. The eukaryotic expression vector pEGFP-hTERT was constructed and transfected into SW579 cells to observe the changes of cell growth status. Detection of telomerase activity. Results The proliferation rate of SW579 cells stably transfected with pEGFP-hTERT was 148%, 48% higher than that of non-transfected cells and 45% higher than that of empty plasmid group (P <0.01). The recombinant plasmids transfected The percentage of cells in G1, S, G2 / M phase was 68.05%, 27.66% and 10.45%, respectively, which were significantly different from those in non-transfected and empty plasmid groups (P <0.01, P <0.05) Proliferation index (PI) of plasmid transfection group was 38.11, which was significantly higher than that of empty plasmid group (14.22) and non-transfected group (13.60). Conclusion Up-regulation of human telomerase expression can promote the proliferation of SW579 cells.