层黏素受体靶向RNA干扰质粒载体的构建  

Study on the construction of LNR-targeting siRNA-expressing plasmid

作　　者：高占峰[1] 李天宇[2] 王曙光[3] 

机构地区：[1]内蒙古医学院附属医院普外科,内蒙呼和浩特010059 [2]北京世桥生物制药有限公司,北京100070 [3]第三军医大学西南医院全军肝胆外科研究所,重庆400038

出　　处：《临床和实验医学杂志》2012年第3期161-163,F0003,共4页Journal of Clinical and Experimental Medicine

基　　金：国家自然科学基金资助(30772109)

摘　　要：目的构建层黏素受体RNA干扰质粒,为探讨抑制层黏素受体过表达在胆管癌免疫逃逸中的作用奠定基础。方法 PCR法获得U6启动子序列以及产生siRNA的相应DNA模板序列,将其克隆入pGenesil-3获得RNA干扰载体pGenesil-shLNR。脂质体法将pGenesil-shLNR导入胆管癌细胞QBC939,免疫组化法检测对胆管癌细胞QBC939LNR表达的抑制情况。结果所构建的载体pGenesil-shLNR能够干扰QBC939细胞中LNR的表达,并且具有新霉素抗性,能够用G-418筛选稳定转化的细胞株。结论成功构建了抑制胆管癌细胞QBC939表达LNR的RNA干扰质粒载体。Objective To construct LNR-targeting siRNA-expressing plasmid,and to explore the significance of LNR inhibition in immune escape of cholangiocarcinoma.Methods U6 promoter cassette and siLNR sequence were obtained by PCR technique and cloned into pGenesil-3 to produce plasmid pGenesil-shLNR,which was transfected into QBC939 cells with limpofectin 2000,and LNR expression was detected by immunohistochemical method.Results The plasmid pGenesil-shLNR could efficiently reduce the expression of LNR and confer G-418 resistance on QBC939 cells.Conclusion The successful construction of siRNA expressing plasmid will facilitate the application of RNA interference technique and lay foundation for further studies on therapeutic effect of LNR inhibition in immune escape of cholangiocarcinoma.

关 键 词：层黏素受体 RNA干扰 质粒 载体构建 

分 类 号：R735.8[医药卫生—肿瘤]