oct4-shrna慢病毒载体沉默结肠癌oct4基因 并诱导细胞凋亡的初步 .doc

Oct4-shRNA慢病毒载体沉默结肠癌Oct4基因 并诱导细胞凋亡的初步研究 文坤明1，张贵海2，陈旺盛1，钱江1，傅仲学1(1.400016重庆，重庆医科大学附属第一医院胃肠外科；2.563003遵义，遵义医学院附属医院肿瘤科) [Alpha 摘要：目的　探讨对癌细胞的抑制效率及凋亡的影响。方法　4对针对Oct4shRNA的，检测的表达Oct4-shRNA慢病毒载体Ｔ包装SW480细胞（内源验证），实时荧光定量和检测癌细胞内Oct4的表达；测定细胞凋亡。结果　Oct4-shRNA慢病毒载体Oct4的和蛋白表达；与对照组相比，细胞凋亡明显增加（）。结论　Oct4-shRNA可有效抑制Oct4表达，并显著增加细胞凋亡。 Oct4；慢病毒载体；细胞凋亡 [中图分类号]：R735.3+5 文献标识码：Ａ Oct4-shRNA lentiviral vectors silences colon cancer Oct4 gene and induces cell apoptosis Wen Kunming1, Zhang Guihai2, Chen Wangsheng1, Qian Jiang1, Fu Zhongxue1(1Department of Gastrointestinal Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400016,China;2 Department of Oncology, Affiliated Hospital of Zunyi Medical College, Zunyi 563003, China) [Abstract]: Objective To investigate the inhibition efficiency of Oct4 by lentiviral vector- mediated RNA interference and the effect on cell apoptosis in colon cancer cell line. Method Overexpression lentiviral vector plasmid of Oct4-GFP fusion and four Oct4-shRNA lentiviral vector plasmids was constructed, and each of the four Oct4-shRNA lentiviral vectors and Oct4-GFP overexpression plasmid were co-transfect respectively into 293T cells, then GFP protein expression in the cells was detected by western bolt（exogenous sieve target）. Through the previous experiment, it was determined which one of the four Oct4-shRNA lentiviral vectors was the best one in interference effect, then the Oct4-shRNA lentiviral vector was transfected into the packaging cells 293T, and at last the supernatant with virus was collected to transfect SW480 cells(endogenous authentication). Quantitative real-time fluorescent PCR and Western-blot were used to detect the expression of Oct4, and flow cytometry was used to detect cancer cell apoptosis. Result To filters out the best interference effect of Oct4-shRNA lentiviral vectors by exogenous screen target, after packaging by 293T cells, the Oct4-shRNA lentiviral vector was transfected into SW480 cell. RNAi gro