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张蕾, 田力, 李伟伟, 李鹏飞. 端粒酶上调与甲状腺鳞癌细胞增殖关系[J]. 中国公共卫生, 2010, 26(11): 1411-1413. DOI: 10.11847/zgggws2010-26-11-34
引用本文: 张蕾, 田力, 李伟伟, 李鹏飞. 端粒酶上调与甲状腺鳞癌细胞增殖关系[J]. 中国公共卫生, 2010, 26(11): 1411-1413. DOI: 10.11847/zgggws2010-26-11-34
ZHANG Lei, TIAN Li, LI Wei-wei, . Upregulation of hTERT expression influence prolification of SW579 cells[J]. Chinese Journal of Public Health, 2010, 26(11): 1411-1413. DOI: 10.11847/zgggws2010-26-11-34
Citation: ZHANG Lei, TIAN Li, LI Wei-wei, . Upregulation of hTERT expression influence prolification of SW579 cells[J]. Chinese Journal of Public Health, 2010, 26(11): 1411-1413. DOI: 10.11847/zgggws2010-26-11-34

端粒酶上调与甲状腺鳞癌细胞增殖关系

Upregulation of hTERT expression influence prolification of SW579 cells

  • 摘要: 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。

     

    Abstract: ObjectiveTo upregulate human telomerase reverse transcriptase(hTERT)expression in SW579 cells and to observe the cell proliferation and telomerase activity.MethodsFull DNA of 3 453 bp human hTERT were obtained with restricted enzyme method and cloned into pEGFP-N1 to reconstruct a recombinant plasmid named pEGFP-hTERT.After transfected into SW579 cells,the cell growth condition and telomerase activity were detected with methyl thiazolyl tetrazolium(MTT),flow cytometry(FCM)and TRAP-PCR method.ResultsThe proliferation rate of SW579 cells of pEGFPhTERT stably transfected was 148%,which increased significantly by 48% than the non-transfection group(P<0.01).The cell proliferation index of the transfection group was 2.8 times than that of non transfection group(P<0.01).The cell numbers of S phase and G2/M phase were 27.66% and 10.45%,respectively,which increased and the the telomerase activity was up regulated too.ConclusionUpregulation of hTERT expression could advance SW579 cells proliferation.

     

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