慢病毒介导的CBS基因沉默对人胶质瘤细胞增殖的影响  

Effect of lentivirus-mediated CBS gene silencing on proliferation of glioma cells

作　　者：闵建平[1] 龙彦伊 王海涛[1] 朱公建[1] 张永东[1] 周海红[1] 王涛[1] 郭红云[1] 苏海翔[1] Min Jianping;Long Yanyi;Wang Haitao;Zhu Gongjian;Zhang Yongdong;Zhou Haihong;Wang Tao;Guo Hongyun;Su Haixiang(Gansu Provincial Academic Institute for Medical Research/Gansu Provincial Cancer Hospital,Lanzhou 730050,China;Northwest Normal University,Lanzhou 730070,China)

机构地区：[1]甘肃省医学科学研究院/甘肃省肿瘤医院,甘肃兰州730050 [2]西北师范大学,甘肃兰州730070

出　　处：《甘肃医药》2023年第3期193-196,共4页Gansu Medical Journal

基　　金：甘肃省卫生行业科研项目(编号:GWGL2014-51、GSWSKY-2019-96);兰州市科技计划指导性项目(编号:2020-ZD-52)。

摘　　要：目的:探究催化产生内源性H2S的酶CBS在人胶质瘤细胞增殖中的作用。方法:以人脑胶质瘤U87细胞为实验对象,使用CBS基因shRNA慢病毒载体转染胶质瘤细胞CBS基因,利用0.5μg/mL嘌呤霉素筛选建立CBS-RNAi慢病毒的U87稳定细胞株,通过qPCR法对各组CBS基因的表达量检测,确定目标基因的沉默效果以及CBS基因沉默对于U87细胞的增殖能力的影响。结果:与空白对照组相比,CBS-RNAi 2组的CBS mRNA表达量明显降低,CBS-RNAi 1组和CBS-RNAi 3组的CBS mRNA表达量降低;证明慢病毒介导的RNAi可有效沉默人胶质瘤U87细胞的CBS基因,且CBS-RNAi组沉默人胶质瘤细胞CBS基因的效果最佳。结论:沉默CBS基因能够抑制人胶质瘤U87细胞的体外增殖,CBS基因或可成为临床治疗人脑胶质瘤的新靶点。Objective:This topic explores the role of enzyme CBS that catalyzes the generic H2S in the proliferation of glioma cells.Methods:Human brain glioma U87 cells are taking as the experimental material,the CBS gene shRNA lentivirus vectors are using for transferring glioma cells,0.5μg/mL puromycin is using for establishing CBS-RNAi lentivirus transferred U87 stable transgenitc strain.The expression levels of the CBS gene of each group is detected by qPCR method and meanwhile the CBS-RNAi silencing effect and its impact on the proliferation of U87 cells is observed.Results:Compare with the control group,mRNA expression level of the CBS-RNAi 2 group is significantly reduced while the mRNA expression level of CBS-RNAi 1 group and CBS-RNAi 3 group is also decreased;it proves that the CBS gene of glioma U87 cells can be effectively silenced by lentivirus mediated CBS-RANi,and the CBS-RNAi 2 group is the best designed compared with the other two groups.Conclusion:The proliferation of human brain glioma U87 cells can be inhibited by CBS gene silencing in vitro,and the CBS gene may become a new target for clinical treatment of human brain glioma.

关 键 词：胶质瘤 硫化氢 CBS 慢病毒 RNA干扰 

分 类 号：R739.41[医药卫生—肿瘤]