siRNA沉默COX-2基因对骨肉瘤MG-63细胞凋亡和自噬的影响  

Effect of COX-2 gene silencing by siRNA on apoptosis and autophagy of osteosarcoma MG-63 cells

作　　者：陈秀锦[1] 杨明宣 崔兆辉[1] 胡旭昌[1] 汪静[1] 安丽萍[1] 马靖琳[1] 王栓科[1] 

机构地区：[1]兰州大学第二医院骨科、甘肃省骨关节疾病研究重点实验室,甘肃兰州730000

出　　处：《肿瘤》2014年第5期425-429,共5页Tumor

基　　金：甘肃省自然科学基金资助项目(编号:1010RJZA124)

摘　　要：目的 :探讨小干扰RNA(small interfering RNA,siRNA)抑制环氧合酶-2(cyclooxygenase-2,COX-2)基因的表达对骨肉瘤MG-63细胞凋亡和自噬的影响及其可能的机制。方法 :将靶向COX-2基因的siRNA(COX-2siRNA)转染至骨肉瘤MG-63细胞后,FCM法检测其对细胞凋亡的影响,实时荧光定量PCR法检测细胞中COX-2、Bcl-2、微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)和Beclin 1 mRNA的表达,蛋白质印迹法检测COX-2、Bcl-2和Beclin 1蛋白的表达。结果 :COX-2 siRNA转染组MG-63细胞中COX-2 mRNA和蛋白的表达水平明显低于阴性对照组(转染control siRNA)和空白对照组(未转染的MG-63细胞)(P<0.05)。COX-2 siRNA转染组MG-63细胞凋亡率明显高于阴性对照组和空白对照组(P<0.05)。COX-2siRNA转染组MG-63细胞中抗凋亡因子Bcl-2 mRNA及其蛋白的表达水平下调(P<0.05),自噬相关因子LC3和Beclin 1 mRNA及Beclin 1蛋白的表达水平上调(P<0.05)。结论 :沉默COX-2基因可以促进骨肉瘤MG-63细胞的凋亡和自噬。Objective： To investigate the effect of cyclooxygenase-2 （COX-2） gene silencing by small interfering RNA （siRNA） on apoptosis and autophagy of osteosarcoma MG-63 cells and explore its possible mechanism, Methods： After COX-2 siRNA which targeting COX-2 gene was transfected into osteosarcoma MG-63 cells, the apoptosis was detected by flow cytometry （FCM）, the expression levels of COX-2, Bcl-2, microtubule-associated protein 1 light chain 3 （LC3） and Beclin 1 mRNAs were measured by real-time fluorogenic quantitative-PCR, and the expression levels of COX-2, Bcl-2 and Beclin 1 proteins were determined by Western blotting. Results： The expression levels of COX-2 mRNA and protein in MG-63 cells after transfection with COX-2 siRNA were lower than those in the negative control cells （MG-63 cells transfected with a control siRNA） and the blank control cells （MG-63 cells without any transfection） （P 〈 0.05）. The apoptosis rate of MG-63 cells after transfection with COX-2 siRNA was higher than that of the negative control cells and the blank control cells （P 〈 0.05）. The expression levels of anti-apoptosis factor Bcl-2 mRNA and protein in MG-63 cells after transfection with COX-2 siRNA were down-regulated （P 〈 0.05）. The expression levels of autophagy-related LC3 and Beclin 1 mRNAs and the Beclin 1 protein in MG-63 cells after transfection with COX-2 siRNA were up-regulated （P 〈 0.05）. Conclusion： Silencing of COX-2 gene may promote the apoptosis and autophagy of osteosarcoma MG-63 cells.

关 键 词：骨肉瘤 RNA干扰 细胞凋亡 自噬 细胞 MG-63 

分 类 号：R738.1[医药卫生—肿瘤]