敲除Ob-R基因后对人乳头状甲状腺癌表达的影响-济宁医学院学报.docVIP

RNAi介导敲除ObR基因后对人乳头状甲状腺癌细胞K1表达的影响* 王帅1,2，孔霞3 ，张国安4，侯森5，崔文5△ (1济南大学山东省医学科学院医学与生命科学学院，山东 济南 250062，2山东省医学科学院，山东 济南 250022；3济宁市第一人民医院药剂科，山东 济宁 272067；4济宁医学院基础学院，山东 济宁 272067；5济宁医学院法医学与医学检验学院，山东 济宁 272067。) 摘 要 目的 探讨用RNAi技术敲除ObR基因后对k1细胞的影响。方法 使用siRNA干扰技术瞬时转染构建立siRNA-ObR。实验设计空白对照组、阴性对照组和siRNA-ObR组。瞬时转染后，Real-time PCR和Western Blotting在mRNA水平和蛋白水平检测siRNA-ObR是否有效抑制K1细胞内靶基因的表达；MTT实验研究ObR沉默后K1细胞的增殖能力；Transwell实验检测ObR沉默后K1细胞的侵袭能力。结果 Real-time PCR实验结果显示，ObR基因沉默后K1细胞在mRNA水平上表达降低；Western Blotting实验结果显示，ObR基因沉默后K1细胞在蛋白水平上有靶向抑制作用。MTT实验结果显示ObR对K1细胞的增殖没有意义；Transwell实验结果显示ObR沉默后K1细胞侵袭能力下降。结论 ObR基因沉默后有效抑制人乳头状甲状腺癌K1细胞内靶基因的表达，降低其侵袭能力。 关键词 RNAi；基因沉默；siRNA；人乳头状甲状腺癌K1细胞；ObR基因 中图分类号:R736.1 文献标识码:A 文章编号： Effect on the expression of hunman papillary thyroid carcinoma K1 cells after RNAi mediated knockdown WANG shuai, ZHANG Guoan, HOU sen, KONG xia, CUI wen (University of Jinan Shandong Academy of Medical Sciences, Jinan 250022, China) Abstract: Objective To study the effect of RNAi-mediated OBR gene silencing on human papillary thyroid carcinoma cell K1. Method siRNA interference technology transient transfection was used to construction of siRNA-ObR. The three groups including control group, negative control group, siRNA-ObR group. After transfection, to detect whether siRNA-ObR inhibit the expression of target genes in mRNA level and protein level. MTT and Transwell respectively detect proliferation and invasion of K1 cells after OBR gene knockout. Result Real-time PCR results show that the level of mRNA expression decreased on K1 cells after ObR gene silencing. Western Blotting results show that the level of protein expression has targeted inhibition on K1 cells after ObR gene silencing. MTT results show that the silencing of ObR were no significance on the proliferation of K1 cells. Transwell results show that the silencing of ObR decreased K1 cells invasion. Conclusion OBR siRNA can effectively inhibit the expression of target gene and can reduce invasion ability on human