Arp2/3复合物表达沉默对胶质瘤细胞侵袭和迁移的影响  

Effect of silencing Arp2/3 complex expression on invasion and migration of glioma cells

作　　者：刘志峰[1] 杨学军[1] 刘彬[1] 陈聪[1] 任炳成[1] 王垒垒[1] 赵恺[1] 于圣平[1] 明浩朗[1] 朱蒙[1] 

机构地区：[1]天津医科大学总医院神经外科,天津300052

出　　处：《中国神经精神疾病杂志》2013年第3期129-134,共6页Chinese Journal of Nervous and Mental Diseases

基　　金：国家自然科学基金项目(编号:81272782)资助

摘　　要：目的研究RNA干扰技术沉默Arp2/3复合物后对胶质瘤细胞片状伪足以及运动能力的影响。方法脂质体介导Arp3-siRNA转染体外常规培养的U251胶质瘤细胞,同时设siRNA-NC(阴性对照组,n=3)组和siR-NA-N(空白对照组,n=3)组。RT-PCR和western blot方法验证三组中Arp3 mRNA和蛋白的表达情况。免疫荧光检测不同处理组的Arp2/3复合物定位以及细胞形态变化。Transwell细胞侵袭实验和划痕实验检测三组细胞的运动能力。结果三组细胞的Arp3 mRNA相对表达水平分别为:19.33%±2.08%、97.33%±2.38%和96.67%±2.52%(P<0.01);三组细胞蛋白相对表达量分别为20.00%±3.00%、85.33%±3.21%和84.33%±4.04%(P<0.01)。与siRNA-N组比较,siRNA-Arp3组胶质瘤细胞Arp3 mRNA和蛋白的表达水平下调分别达80%和76%。免疫荧光结果显示siRNA-Arp3组细胞较siRNA-NC组和siRNA-N组细胞片状伪足明显变小甚至消失。Transwell细胞侵袭实验和划痕实验显示siRNA-Arp3组胶质瘤细胞运动能力较siRNA-N组分别下降达69%(P<0.01)和68%(P<0.01)。结论 Arp2/3复合物通过影响片状伪足的形成,从而在胶质瘤细胞的运动过程中发挥重要作用,提示Arp2/3复合物可作为治疗脑胶质瘤的一个候选靶点。Objective To investigate the inhibitory effect of Arp2/3 complex silenced by siRNA on lamellipodia and motility capability of human glioma cells in vitro. Methods The Arp3-siRNA and non-sipencing SiRNA were transfeeted into human glioma cells U251 using oligofectamine, respectively. Human glioma ceils were then divided into siRNA siR- NA- negative control treated group （siRNA-NC） and non-silencing siRNA treated groups （siRNA-N）. RT-PCR and west- ern blotting assay were used to detect the mRNA and protein expression of Arp3. Immunofluoreseenee was employed to ex- amine the location of Arp2/3 complex in glioma cells and the morphological changes of glioma cells. Transwell invasion as- say and the wound-healing assay were used to determine the invasion and migration of U251 glioma cells. Results The mRNA expression levels of Arp3 in siRNA, siRNA-NC and siRNA-N groups were 19.33% ± 2.08%, 97.33% ±2.38%, and 96.67% ± 2.52%, respectively （P〈 0.01）. The protein expression levels in siRNA, siRNA-NC and siRNA-N groups were 20.00% ± 3.00%, 85.33% ± 3.21%, and 84.33%±4.04%, respectively （P〈0.01）. The mRNA and protein expres- sion levels of Arp3 in Arp3-siRNA treated U251 cells was significantly inhibited up to 80% and 76% compared with that in siRNA-N treated group. The lamellipodia became small or disappeared in siRNA treated U251 glioma cells whereas were kept normal in siRNA-N treated group and siRNA-NC treated group. The invasiveness and migration capability of glioma cells in the Arp3-siRNA treated group were decreased up to 69% （P〈0.01） and 68% （P〈0.01） compared with the siRNA-N treated group. Conclusions The Arp2/3 complex plays an important role in the spread of glioma cells through the lamellipodia, which suggests that Arp2/3 complex might be a good candidate for anticancer strategy to prevent the inva- sion and migration of glioma cells.

关 键 词：胶质瘤 片状伪足Arp2/3复合物 RNA干扰 

分 类 号：R739.4[医药卫生—肿瘤]