RNAi沉默Polo-like kinase-1基因表达对胰腺癌细胞增殖的影响  

Effects of polo-like kinase-1 gene silence by RNA interference on proliferation of human pancreatic cancer cells

作　　者：张尤历[1] 范钰[1] 吴莺[1] 张宇川[1] 张炜[1] 

机构地区：[1]江苏大学附属医院消化科,江苏省镇江市212001

出　　处：《世界华人消化杂志》2007年第33期3484-3488,共5页World Chinese Journal of Digestology

摘　　要：目的:探讨polo-like kinase-1(PLK1)基因在胰腺癌细胞中的作用.方法:采用PLK1小干扰核糖核酸分子(small interfering RNA,siRNA)转染人胰腺癌Mi- aPaCa-2细胞后,分别采用实时定量PCR和Western blot检测PLK1基因mRNA和蛋白表达水平,观察PLK1 siRNA转染对胰腺癌细胞体内外增殖的影响.于转染不同时间后收集细胞,分别采用琼脂糖凝胶电泳和TUNEL方法检测胰腺癌细胞凋亡情况.结果:胰腺癌MiaPaCa-2细胞经siRNA转染处理后,PLK1 mRNA和蛋白表达水平明显下降(P<0.05).PLK1基因siRNA可明显抑制癌细胞体外生长(P<0.05)和体内裸鼠模型增殖(P<0.05).细胞凋亡检测发现,DNA电泳出现明显的梯度图谱,且与浓度相关(r=0.836,P<0.05).TUNEL结果显示,转染组癌细胞凋亡指数明显增加,且呈时间和浓度依赖性(r= 0.875,P<0.05).结论:PLK1 siRNA转染可明显抑制胰腺癌细胞增殖,其机制可能与诱导细胞凋亡有关.AIM： To investigate the role of polo-like kinase-1 （PLK1） in human pancreatic cancer cells. METHODS： After MiaPaCa-2 pancreatic cancer cells were transfected with small interfering RNA （siRNA） against PLK1, real-time RT-PCR and Western blotting were used to examine PLK1 gene expression in all cancer cells. The proliferation and growth of cancer cells in vivo and in vitro were studied. Apoptosis of cancer cells was evaluated by terminal uridine deoxynucleotidyl nick end labeling （TUNEL） and aga- rose gel electrophoresis, respectively. RESULTS： Expression of PLK1 in MiaPaCa-2 cancer cells transfected with siRNA was downregulated significantly. Transfection of PLK1 siRNA resulted in significant inhibition of pancreatic cancer cells in vivo and in vitro. The results from TUNEL and DNA ladder analysis showed cancer cells exhibited marked apoptosis,in a time- and dose-dependent manner （r = 0.875, P 〈 0.05）. CONCLUSION： RNA interference PLK1 can inhibit proliferation through inducing apoptosis of human pancreatic cancer cells.

关 键 词：胰腺肿瘤 PLK1 RNA干扰 小干扰RNA增殖 凋亡 实时定量PCR 免疫印迹 

分 类 号：R735.9[医药卫生—肿瘤]