敲低Plk1对人肝癌血管内皮细胞迁移影响研究  

Influence of knocking down Plk1 on migration of human hepatocellular carcinoma tumor-derived endothelial cells

作　　者：于慧杰[1] 吴晔[2] 李峰生[1] 李伟[1] 江其生[1] 

机构地区：[1]第二炮兵总医院中心实验室,北京100088 [2]中国人民解放军总医院血管外科,北京100853

出　　处：《中华肿瘤防治杂志》2015年第8期584-587,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81172130;81202151)

摘　　要：目的研究人Polo样激酶1(polo-like kinase 1,Plk1)在人肝癌血管内皮细胞(human hepatocellular carcinoma tumor-derived endothelial cell,TEC)迁移过程中的作用。方法用含有20%胎牛血清的RPMI1640培养基培养TEC细胞,合成Plk1和阴性对照小干扰片段。实验设置Plk1siRNA处理组和siRNA阴性组(对照组)。分别用Plk1siRNA和阴性对照siRNA转染TEC,采用蛋白质印迹法验证Plk1siRNA干扰能否有效降低TEC细胞的Plk1蛋白表达水平;采用Transwell细胞迁移实验和细胞划痕实验评估Plk1siRNA转染后TEC迁移能力的变化。结果蛋白质印迹法检测结果显示,Plk1siRNA转染后TEC细胞中Plk1蛋白表达降低。Transwell细胞迁移实验发现,Plk1siRNA处理组12h穿膜细胞数为130.9±23.0,低于siRNA阴性组的184.9±26.6,t=-5.943,P<0.001。细胞划痕实验显示,Plk1siRNA处理组细胞12h迁移距离为(17.5±7.0)像素,低于siRNA阴性组的(72.6±12.4)像素,t=-6.698,P=0.006;Plk1siRNA处理组细胞24h迁移距离为(70.9±6.9)像素,低于siRNA阴性组的(137.9±18.9)像素,t=-5.761,P=0.016。结论敲低Plk1能显著抑制人肝癌血管内皮细胞的迁移能力,提示Plk1可能在人肝癌血管内皮细胞转移中具有一定作用。OBJECTIVE To investigate the role of Polo-like Kinase l（Plkl） in migration of human hepatocellular carcinoma tumor-derived endothelial cells （TEC）. METHODS TEC cell line was cultured in RPMI1640 with 20% fetal bovine serum, siRNA segments targeting Plkl and negative control siRNA were synthesized. The PlklsiRNA group and siRNA negative control group were set in this experiment, siRNA segments targeting Plkl and negative control siRNA were transfected into TEC cells. Effect of PlklsiRNA on the downregulation of Plkl protein expression in TEC was evalu- ated by Western blot. After transfection, the changes in migration of TEC were measured by wound healing and Transwell migration assay. RESULTS Successful transfection was confirmed using Western blot. Plkl expression at the protein level in TEC was reduced by the PlklsiRNA segments. In transwell migration assay, the number of TEC crossing through chambers in PlklsiRNA group （130.9±23.0） was lower than that in the siRNA control group（184.9± 26.6） ,t= -5. 943,P〈0. 001. In wound healing assay, the migration distance of Plkl siRNA group was （17.5 ± 7.0） Pixel in 12 h, which was lower than that in the siRNA control group（72.6±12.4） ,t= -6. 698,P=0. 006; and the migration distance of Plkl siRNA group was （70.9±6.9） Pixel in 24 h,which was lower than that in the siRNA control group（137.9±18.9） ,t= -5. 761 ,P=0. 016. CONCLUSION Plkl knocking down by siRNA may inhibit the migration of TEC, suggests that Plkl might play an important role in the metas- tasis of human hepatocellular carcinoma tumor-derived endothelial cells.

关 键 词：血管内皮细胞 POLO样激酶1 RNA干扰 细胞迁移 肝肿瘤 

分 类 号：R735.7[医药卫生—肿瘤]