ATM基因_____ATM基因；_RNA干扰；宫颈癌；转染

ATM基因siRNA真核表达载体的构建及其对转染宫颈癌细胞株ATM表达的抑制作用

http://www.100md.com 《第四军医大学学报》 2005年第5期

ATM基因,,ATM基因；,RNA干扰；宫颈癌；转染,1材料和方法,2结果,3讨论,【参考文献】

     Construction of eukaryotic expression plasmid expressing siRNA targeting ATM gene and its inhibitory effect on ATM expression in transfected human cervical carcinoma cells

    WEI Li1, XIN XiaoYan1, WANG Jian1, LEI YingFeng2, XUE XiaoPing2

    1Department of Obstetrics & Gynecology, Xijing Hospital, 2Department of Microbiology, School of Basic Medicine, Fourth Military Medical University, Xian 710033, China

    【Abstract】 AIM: To study inhibitory effect of siRNA on ATM expression in human cervical carcinoma cells and to determine the feasibility of RNAi application in genetherapy of cervical cancer. METHODS: Hairpin siRNA templates were designed based on ATM gene sequence and mRNA structure and was cloned into pSuppressor vector. Siha cells were transfected by pSuppressorATM with electroporation and the nontransfected cells and nonspecific siRNA transfected cells were used as controls. The inhibitory effect of ATM mRNA was detected by semiquantitative RTPCR and Westernblot, and the inhibitory effect of ATM protein expression was detected by FCM and immunofluorescency. RESULTS: Semiquantitative RTPCR, Westernblot, FCM and immunofluorescency all revealed a potent inhibition of the expression of ATM in the transfected Siha cells. CONCLUSION: The plasmid vector was successfully constructed. Transient transfection can significantly decrease the ATM level in transfected cells. ......

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