RNA干扰靶向沉默FLIP基因对卵巢癌细胞A2780生物学特性的影响及生长抑制作用  

Effect of FLIP-siRNA-mediated gene silencing on bionomics and growth inhibition in the ovarian cancer cell line A2780

作　　者：王毓[1] 刘培淑[1] 毛洪鸾[1] 周黎光[1] 孙玉兰[2] 姜虹[3] 王荣[3] 

机构地区：[1]山东大学齐鲁医院妇产科,济南250012 [2]新泰市人民医院产科,山东泰安271200 [3]教育部和卫生部心血管重构与功能研究重点实验室,济南250012

出　　处：《山东大学学报（医学版）》2008年第2期154-158,共5页Journal of Shandong University:Health Sciences

摘　　要：目的探讨靶向FLIP基因的小干扰RNA(siRNA)对人卵巢癌细胞A2780生物学特性的影响及生长抑制作用。方法设计并体外化学合成FLIP序列特异性双链RNA,在脂质体(LipofectamineTM2000)介导下转染人卵巢癌细胞株A2780。采用半定量RT-PCR和Western blot法检测FLIP-siRNAs转染前后A2780细胞FLIP基因mRNA和蛋白表达的变化,并筛选出抑制作用最强的FLIP-siRNA。采用四甲基偶氮唑盐(MTT)法、Annexin-V-PI双染法流式细胞术(FCM)分别检测FLIP-siRNA转染对A2780细胞的生长抑制作用和对凋亡的影响。结果特异性FLIP-siRNAs片段能有效降低A2780细胞中FLIP的mRNA和蛋白水平(P<0.01),最大抑制率分别为77.4%和66.1%;转染FLIP-siRNA后,A2780细胞的生长活力明显下降(P<0.05),RNA干扰组的细胞凋亡也显著增加(P<0.05)。结论靶向FLIP基因的siRNAs可在转录和翻译水平抑制FLIP基因表达;并可抑制卵巢癌A2780细胞生长,促进其凋亡。Objective To investigate the effects of FLIP-siRNA-mediated gene silencing on bionomics and growth inhibition in the ovarian cancer cell line A2780. Methods Based on the sequence of FLIP mRNA, three siRNAs were designed and chemically synthesized. They were transfected into the ovarian cancer cell line A2780 with Lipofectamine^TM 2000. The FLIP mRNA and protein levels were determined by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot. The FLIP-siRNA having the most powerful inhibition was selected. Cell growth inhibition and apoptosis were determined by M＇IT assay and flow cytometry. Results Short siRNAs targeting FLIP down-regulated the mRNA and the protein level of the FLIP oncogene（ P 〈 0.01）, and the inhibition rate reached to 77.4% and 66.1%, respectively. The proliferation of the ovarian cancer cell line A2780 was inhibited（ P 〈 0.05） and the ratio of apoptesis significantly increased after transfection （ P 〈 0.05）. Conclusion FLIP-siRNAs can significantly down-regulate FLIP expression in transcriptional and translational levels, inhibit cell growth and induce cell apoptosis in the ovarian cancer cell line A2780.

关 键 词：RNA干扰 双链RNA 基因 FHP 卵巢肿瘤 

分 类 号：R737.31[医药卫生—肿瘤]