RNAi干扰Shp2基因对K562细胞增殖的抑制效应  

The Inhibitory Effect of RNAi Shp2 Gene on the Proliferation of Chronic Myeloid Leukemia K562 Cells

作　　者：王灿蔚[1] 陶崑[1] 齐杰玉[1] 邓一平[2] 

机构地区：[1]重庆医科大学基础医学院免疫学教研室重庆医科大学分子医学与肿瘤研究中心,重庆40016 [2]重庆医科大学实验中心,重庆40016

出　　处：《中国细胞生物学学报》2014年第1期75-80,共6页Chinese Journal of Cell Biology

基　　金：重庆市科委(批准号:CSTC:2010BB5369)资助的课题~~

摘　　要：通过RNA干扰技术沉默蛋白酪氨酸磷酸酶Shp2基因,构建重组质粒,采用实时荧光定量PCR(Real-time PCR)法、Western blot、MTT法、流式细胞术(FCM)分别检测转染后K562细胞中bcr/abl融合基因、bcr/abl融合蛋白的表达水平、细胞生长增殖变化及细胞凋亡率,探索该基因的沉默表达对K562细胞的抑制作用。结果表明,该实验成功构建出能明显下调Shp2基因及其蛋白表达的重组质粒,转染K562细胞后,其bcr/abl融合基因及融合蛋白水平均明显降低、K562细胞增殖活力被抑制(P<0.05)、细胞凋亡水平上升(P<0.05)。与对照组相比,其差异具有统计学意义。提示,重组质粒可显著降低bcr/abl基因及蛋白的表达,抑制K562细胞的生物学效应,表明在细胞水平沉默Shp2有可能成为治疗慢性粒细胞白血病的有效靶点。We established low-expressed Shp2 recombinant plasmids containing siRNA fragment specific for Shp2 gene. Real-time PCR and Western blot assays were used to detect the mRNA and protein level of bcr/abl gene. The proliferation of cells was detected by MTT assay, and the apoptosis was detected by Flow Cytometry （FCM）. The results demonstrated that the recombinant plasmids were successfully constructed and significantly inhibited the expression of Shp2 gene. Compared with the two control groups, the mRNA and protein level of bcr/abl were significantly decreased; the proliferation ability was inhibited significantly （P〈0.05） and the rate of apoptosis cells was increased significantly （P〈0.05）. The biological effect of K562 cells may be significantly inhibited by down-regulation of Shp2, indicating that Shp2 may play a biological role as a tumor suppressor gene.

关 键 词：慢性粒细胞白血病(CML) BCR ABL Shp2 K562细胞 

分 类 号：Q78[生物学—分子生物学]