BubR1基因表达对肝癌细胞对紫杉醇的敏感性的影响

Downregulation of BubR1 gene expression enhancesthe sensitivity of hepatocellular cells to paclitaxel

目的:探讨BubR1基因在紫杉醇治疗肿瘤中的作用。方法:构建针对BUBR1基因的特异性短发夹RNA载体,导入人肝癌细胞株HepG-2,转染48 h后,运用实时荧光定量PCR、Western印迹法评价RNA干扰效果;BubR1表达下调的肝癌细胞及对照组细胞用不同浓度(1、3、10、30、100、300、1000和3000 nmol/L)的紫杉醇处理,MTT法检测细胞增殖情况。结果:shRNA-BubR1可有效抑制BubR1的mRNA、蛋白水平以及内源性表达。MTT结果显示,BubR1表达下调的细胞在不同浓度紫杉醇作用下的增殖抑制率与对照组相比明显增加(P<0.05),并呈现剂量依赖性。结论:下调BubR1的表达能增强肝癌细胞对紫杉醇的敏感性。

Objective To investigate the role of BubR1 cin the therapeutic effects of paclitaxel ontumor. Methods P1asmid vector containing BubR1 specific shRNA was constructed and eleetroporated into humancervix cancer HepG-2 cells. RFQ-PCR, Western blotting method were used to evaluate the silencing em ciency of RNA interference after 48 h.HepG-2 cells with down-regulated BubR1 and control cells were treated with paclitaxe1 at different concentrations (1,3,10,30,l0o,30o,1 000 and 3 000 nmol/L).The proliferation of cells were measured by MTI1 assay.Results BubR1 specific shRNA effectively inhibited the mRNA and protein expression of BubR1 in HepG-2 cells and endogenous expression.The resultsof MTT indicated that the growth inhibition rate was obvi-ously increased in those cells with down-regulated BubR1 compared with control cells (P<0.05) and the growth inhibition effect was in a dose-dependent manner. Conclusion Down-regulation of BubR1geneexpression can enhance the sensitivity of cervical cancer cells to paclitaxe1.