核转染仪辅助转染肝素酶基因siRNA有效性的实验研究  

Efficacy study of transfection of siRNA of Heparanase gene by nuclear transfetion machine

作　　者：张彩虹[1,2] 董宏伟 肖文华[1] 董伟伟[1] 赵慧霞[1] 段昕妤[1] 李秋文[1] 郝怡鑫[1] 王如良[1] 朱建华[1] 叶明[1] 

机构地区：[1]解放军总医院第一附属医院肿瘤一科 [2]青海大学医学院免疫教研室 [3]66055部队医院

出　　处：《现代仪器与医疗》2013年第5期1-4,共4页Modern Instruments & Medical Treatment

基　　金：国家自然科学基金(编号:30873037)

摘　　要：目的研究核转染仪转染肝素酶基因siRNA的有效性。方法采用核转染仪辅助转染技术和常规脂质体转染技术转染针对肝素酶基因启动子区和编码区设计siRNA,并通过实时定量RT-PCR,Western blot等技术检测2种转染技术转染后,肝癌细胞SMCC-7721在48h、72h和96h时肝素酶基因表达情况。结果 2种转染技术在转染后48h,从mRNA和蛋白水平上均能成功干扰掉肝素酶基因;但在转染后72h,脂质体转染组基因很快恢复表达,而核转染仪转染组基因仍保持沉默;在96h后,2组的肝素酶基因均恢复表达。结论核转染仪可有效转染肝素酶基因siRNA,并使肝癌细胞SMCC-7721的肝素酶基因表达下调,且维持较长时间。Objective To study the effects of transfection of siRNA of heparanase（HPA） gene by nuclear transfection machine and lipofection in liver cancer cell SMMC7721.Methods Several of the siRNA covered promoter region and coding region of HPA gene respectly were designed and synthesed,and then transfected into the hepatoma cells SMMC-7721 by neuclear trancfection.HPA gene expression in both mRNA and protein were detected by RT-PCR,Western blot techniques.Results Both nuclear transfection and lipofection of siRNA of HPA gene can almost completely silence HPA gene at 48h after siRNA transfection.The expression of HPA gene was recovered at 72h after lipofection,but HPA still keep silencing in nuclear transfection group.However,the expressions of HPA gene in both groups were recovered at 96h after gene transfection.Conclusion Nuclear transfection with nuclear transfection machine can effectively silence HPA gene.

关 键 词：核转染仪 肝素酶基因 肝癌细胞 

分 类 号：R735[医药卫生—肿瘤]