中文摘要：

该文研究端粒酶在人结肠癌HCT116高非整倍体变异组及低非整倍体变异组细胞中的表达差异及其端粒酶抑制剂3′-叠氮-3′-脱氧胸苷（3′-Azido-3′-deoxythymidine,AZT）对2组细胞增殖及凋亡的影响。取HCT116细胞加入盐酸强力霉素16 h后撤药,称为高非整倍体变异组;另取HCT116细胞不作任何处理,设为对照组,称为低非整倍体变异组;在撤药后第11 d,采用100、250μmol/L的AZT处理2组细胞72 h,并分别设立空白对照组（未加AZT的高非整倍体变异组及低非整倍体变异组）。采用染色体滴定法进行染色体计数。采用Western blot检测MAD2L1、PUMA、BAX、P21、γ-H2AX蛋白质水平。采用荧光定量PCR检测h TERT、PUMA、BAX、NOXA、P21基因表达,端粒酶活性试剂盒检测端粒酶活性,CCK-8法检测细胞生存率。实验结果表明,采用盐酸强力霉素诱导HCT116细胞的非整倍体率可达到77.33%;高非整倍体变异组细胞h TERT基因的表达及端粒酶活性明显高于低非整倍体变异组;加入AZT后,高非整倍体变异组P21、γ-H2AX蛋白质水平上升程度较整倍体明显,低非整倍体变异组PUMA、BAX蛋白质水平上升程度较高非整倍体变异组明显。该研究表明,盐酸强力霉素可以诱导非整倍体形成,高非整倍体变异组的端粒酶活性及h TERT基因表达高于低非整倍体变异组,AZT可以对高非整倍体变异组和低非整倍体变异组细胞产生增殖抑制作用、DNA损伤作用、细胞周期阻滞作用、诱导凋亡作用。

英文摘要：

The aim of the study was to study the differences of telomerase activities and the h TERT gene expression in high aneuploid variation group and low aneuploid variation group of HCT116 cells and the influence of telomerase inhibitor 3′-Azido-3′deoxythymidine（AZT） on cell proliferation and apoptosis of two group cells. One group of HCT116 cells were added into doxycycline, known as high aneuploid variation group. One group of HCT116 cells were added nothing, known as control group（low aneuploid variation group）. After the withdrawal of doxcycline, the two groups were dealt with 100, 250 μmol/L AZT for 72 hours, control blank group and AZTdealt group were established. The chromosomes numbers of two groups were counted by karyotype analysis. The protein levels of MAD2L1, PUMA, BAX, P21, γ-H2 AX were detected by Western blot. Fluorescent quantitative PCR was used to detect the gene expression of PUMA, BAX, NOXA, P21, h TERT. The telomerase activity of two groups were detected by the Telomeric Repeat Amplification Protocol（TRAP）. CCK-8 was used to detect cell survival after added into AZT. The experiment results showed that the aneuploidy rate of cells could reach to 77.33% while added into doxycycline hydrochloride, the h TERT gene expression and telomerase activity of Dox（＋） group were significantly higher than that of Dox（–） group. The P21, γ-H2 AX protein levels of two groups were increased. The degree of increasing of Dox（＋） group was more obvious. The PUMA, BAX protein expression of Dox（–） group increased after AZT, while it didn＇t increase of Dox（＋） group. It was showed that doxycycline can induce aneuploidy, the h TERT gene expression and telomerase activity of Dox（＋） group were significantly higher than that of Dox（–） group. AZT could inhibit cell proliferation, produce DNA damage and cell cycle arrest of two group cells.