摘要
目的观察人端粒保护蛋白1(human protection of telom eres 1,hPOT1)基因过表达对HeLa细胞染色体端-端融合和端粒酶活性的影响。方法利用先前构建的hPOT1基因真核表达重组质粒pcDNA3-hPOT1,经脂质体介导瞬时转染HeLa细胞介导hPOT1基因过表达;秋水仙碱阻滞细胞周期于分裂中期,DAPI荧光染色分析染色体端-端融合;Telom erase PCR-ELISA试剂盒检测端粒酶活性。结果pcDNA3-hPOT1重组质粒转染HeLa细胞5 d后,染色体端-端融合几率增加,对端粒酶活性无明显影响。结论提示hPOT1蛋白可导致染色体端-端融合。
Objective To observe the effects of hPOT1 overexpression on chromosome end-to-end fusions and telomerase activation in HeLa cells. Methods By an eukaryotic expression plasmid of pcDNA3-hPOT1 constructed preliminarily, liposome mediated transient transfection of HeLa cells and hPOT1 overexpression. Cell cycles were blocked in metaphase by colchicina. Chromosome end-to-end fusions were analyzed by DAPI fluorescein stain. Telomerase PCR-ELISA kit was used to determine telomerase activations. Results After 5 days of the pcDNA3-hPOT1 recombinant plasmids transfected into HeLa cells, the probabilities of chromosome end-to-end fusions inereased. And there was no obvious effect on telomerase activation. Conclusions hPOT1 may induce chromosome end-to-end fusions.
出处
《中国老年学杂志》
CAS
CSCD
北大核心
2006年第8期1052-1054,共3页
Chinese Journal of Gerontology
基金
广东省自然科学基金项目(0401397)
关键词
人端粒保护蛋白
HELA细胞
端-端融合
端粒酶
Human protection of telomeres 1
HeLa cell line
End-to-end fusions
Telomerase