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MicroRNA-mediated NBS1 Gene Silence and Its Effects on Telomerase Activation in Hela Cells
Objective:To research the silence of NBS1 after transfection microRNA expressing eukaryotic recombinants and the changes of telomerase activation in teiomerase-positive cell line Hela.Methods:According to the sequence of NBS1 mRNA,the NBS1 pre-microRNA was designed and synthesized,then cloned into the GFP reporter pcDNA6.2-GW/EmGFP-miR vector and transfected into Hela cells.The integrity of the insert fragment was verified through colony PCR and sequencing analysis.The NBS1 gene expression of NBS1 microRNA recombinants was detected by Real-Time PCR and western blot.Telomerase activity in Hela cells was assayed by TRAP-PCR-EB.Results:Sequences of insert fragment in microRNA expressing recombinants were correct.The NBS1 gene expression was decreased,and the telomerase activation of Hela cell reduced.Conclusion:NBS1 microRNA inhibits NBS1 gene expression,and depresses telomerase activation of Hela cells.This confirms that there is relevance between NBS1 gene and telomerase activity.
作 者： CAO Sun-qiong REN Chang-shan 作者单位： The Second Lab,Tumor Institute,the First Clinical College,China Medical University,Shengyang 110001,China 刊 名： 中国癌症研究（英文版） ISTIC 英文刊名： CHINESE JOURNAL OF CANCER RESEARCH 年，卷(期)： 2008 20(3) 分类号： Q786 关键词： NBS1 gene RNA interfere MicroRNA Telomerase activation