K562细胞中Aurora A与端粒酶表达相关性

Relationship between Expression of Aurora A and Telomerase Activity in K562 Cell

目的探讨人白血病细胞K562中Aurora A与端粒酶表达的相关性,阐明Aurora A在人白血病发生发展中的作用机制。方法用RT-PCR方法克隆得到Aurora A全长基因,采用Fugene6转染试剂介导将其转染入K562细胞中表达,Western Blot检测细胞Aurora A和人类端粒酶反转录酶(hTERT)表达水平,端粒酶检测试剂盒Telomerase PCR ELISA检测细胞端粒酶活性,监测Aurora A表达升高的阳性细胞hTERT和端粒酶活性变化。将针对Aurora A mRNA的DNA核酶导入Aurora A高表达的K562细胞以抑制Aurora A表达,检测Aurora A表达下调后细胞hTERT和端粒酶活性变化。结果Aurora A全长基因转染K562细胞后,各阳性克隆Aurora A表达均上调,并伴hTERT及端粒酶活性增加。DNA核酶转染Aurora A高表达的K562后,能显著下调细胞Aurora A表达,hTERT和细胞端粒酶活性也同步下降。结论在人白血病中,Aurora A表达增加可引起细胞端粒酶活性的同步增加,可能是Aurora A导致白血病发生发展的重要原因。

Objective To explore the relationship between expression of Aurora A and telomerase activity in human leukemia cell,thus elucidate the role Aurora A plays in the carcinogenesis of human leukemia. Methods Full length Aurora A gene was cloned through reverse transeriptase -polymerase chain reaction（ RT- PCR） and transfected into K562 cell by Fugene 6. Western Blot was used to detect the expression level of Aurora A and human telomerase reverse transcriptase（ hTERT）, the telomerase PCR ELISA was used to detect the telomcrase activity of the cells. The variation of hTERT expression and telomerase activity were monitored in cells with increased Aurora A. Meanwhile, DNAzymes targeted to Aurora A mRNA were transfected into K562 cells with high Aurora A level to inhibit their Aurora A expression, the variation of hTERT level and telomerase activity were monitored in cells with down - regulated Aurora A. Results The full length Aurora A gene was transfected into K562 cell, the positive clones with high level of Aurora A over expressed hTERT and telomerase. When K562 - Aurora A was treated with DNAzymes targeted to Aurora A,the Aurora A level was down - regulated, the hTERT level and telomerase activity were reduced simultaneously. Conclusion Over expression of Aurora A activated telomerase expression in human leukemia cell, which may be one of the explanations why Aurora A can induce the carcinogenesis of leukemia.