RPS20 RNA干扰慢病毒的构建及其对CT26细胞生长的影响  

Construction of Lentivirus of RNA Interference for RPS20 and Its Effect on CT26 Cells Growth

作　　者：陈炜璇[1] 高小玲[1] 陶玉珠[1] 李茹柳[1] 郭文峰[1] 陈蔚文[1,2] 

机构地区：[1]广州中医药大学脾胃研究所,广州510405 [2]上海市高校中医内科学E-研究院/上海中医药大学,上海201203

出　　处：《中药新药与临床药理》2012年第3期258-263,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(90809001);广东省中医药局课题(2008432);上海市教育委员会E-研究院建设计划项目(E03008);广州中医药大学中医内科学特色重点学科建设项目

摘　　要：目的研究脾虚证差异表达基因——核糖体蛋白S20(RPS20)RNA干扰慢病毒转染对小鼠结肠癌细胞(CT26)细胞指数的影响,验证前期筛选的RNA干扰序列的有效性,为后续在小鼠体内以RNA干扰进行RPS20的功能鉴定提供参考。方法根据前期筛选的RPS20 RNA干扰有效序列,进行质粒载体的重组,包装RPS20干扰慢病毒并转染CT26细胞,采用xCELLigence细胞动态分析仪实时监测转染后细胞的细胞指数以反映细胞的生长情况。结果测序图谱和Blast比对结果表明重组慢病毒包装载体构建成功,慢病毒滴度为1.2×105TU/mL,RNA干扰慢病毒转染后CT26细胞指数的增长受到明显抑制,转染后30 h细胞指数抑制率达到82.24%。结论前期筛选的RPS20 RNA干扰序列可成功构建包装干扰慢病毒,转染后能有效抑制CT26细胞的生长,可为后续研究提供参考。Objective The effect on cell index of CT26 cells transfected by lentivirus of RNA interference for spleen- deficiency-related gene RPS20 was investigated to verify the validity of previous RNA interference sequence, thus to provide reference for identification of RPS20＇s function by RNA interference in mice. Methods We reconstructed the plasmid vector and produced lentivirus of RNA interference for RPS20 according to the effective RNA interference sequence for RPS20 previously selected. CT26 cells were transfected with the lentivirus and the cell index was moni- tored by xCELLigence real time cell analyzer, which reflected the growth condition of the cells. Results The results of sequencing test and Blast contrast showed the recombinant lentiviral vector was successfully constructed. The titer of the lentiviral stocks was 1.2x105 TU/mL. CT26 cells＇index was suppressed after transfeeted with lentivirus of RNA in- terference and its suppressive rate was 82.24 % 30 h after transfection. Conclusion The previous RNA interference sequence for RPS20 can be constructed into lentivirus successfully and effectively inhibits the growth of transfected CT26 cells, which can provide reference for further reseach.

关 键 词：RNA干扰 CT26细胞 慢病毒 细胞动态分析仪 

分 类 号：R285.5[医药卫生—中药学]