靶向保罗样激酶的小干扰RNA体内外抑制胃癌细胞生长的研究  

Study of Growth inhibition of gastric cancer cells by siRNA targeting polo like kinase 1 in vitro and vivo

作　　者：兰斌[1] 刘炳亚[1] 陈雪华[1] 瞿颖[1] 张晓青[1] 蔡劬[1] 戴起宝[2] 朱正纲[1] 

机构地区：[1]上海交通大学附属瑞金医院外科上海消化外科研究所,200025 [2]福建医科大学附属第一医院肿瘤外科

出　　处：《中华外科杂志》2006年第1期40-44,共5页Chinese Journal of Surgery

基　　金：国家"973"重点基础研究发展规划基金资助项目(2002CB713700)

摘　　要：目的观察敲除保罗样激酶(plk)1基因对胃癌细胞MKN45体内外生长的抑制作用,探讨plk1基因作为胃癌基因治疗靶点的可行性及有效性。方法应用RNA干扰技术(RNAi)抑制MKN45细胞plk1基因的表达,实时定量PCR(real-time quantitative PCR)及蛋白印迹(western blotting)检测plk1mRNA及蛋白质的表达变化,流式细胞仪检测MKN45细胞周期分布及凋亡率的变化,噻唑蓝(MTT)法检测MKN45细胞增殖速度的变化。裸鼠皮下移植plk1小干扰RNA(siRNA)处理的MKN45细胞,观察其成瘤性的改变。western blotting检测成瘤标本的plk1蛋白质的变化。结果MKN45细胞经plk1siRNA作用后,plk1mRNA及蛋白质水平在一定时间内均明显降低,更多的MKN45细胞聚集于G2/M期附近(P<0·05);经plk1siRNA作用的MKN45细胞在48及72h凋亡率高于对照组(P<0·05);生长速度亦明显缓于对照组(P<0·05);在裸鼠体内成瘤性明显减低(P<0·05);但各组体内成瘤标本的plk1蛋白表达无明显改变。结论靶向plk1的siRNA可以抑制胃癌MKN45细胞在体内外的生长速度,plk1有可能成为新的胃癌治疗靶点。Objective To observe the effect of polo like kinase 1 （plkl） gene depletion on the growth of gastric cancer cell line-MKN45 cells in vitro and vivo and discuss the feasibility and effectiveness of arranging plkl as gene therapeutic target for gastric cancer. Methods The plkl expression of MKN45 cells was inhibited by RNA interference （RNAi）. The plkl mRNA and protein level were measured by real- time quantitative PCR and western blotting, and the change of cell cycle distribution and apoptosis rate were detected by flow-cytometry, and the MKN45 cells proliferation was measured by MTT method. MKN45 cells treated with plkl siRNA were transplanted subcutanuously in nude mice and their tumorgenesis ability were observed, the plkl protein levels of the samples from nude mice in different groups were compared. Results After treatment with plkl siRNA, plkl mRNA and protein level decreased obviously in certain time, more MKN45 cells accumulated at G2/M （P 〈 0. 05）. Apoptosis rate of MKN45 cells treated with plkl siRNA was higher than that of control cells at 48 h and 72 h （ P 〈 0. 05 ）, and MKN45 cells proliferated slowerly than control groups（P 〈0. 05）, while the tumorgenesis ability obviously decreased, but the plkl protein levels of the samples from nude mice in different groups were not different. Conclusions siRNA targeting plkl can inhibit the proliferation of MKN45 cells in vitro and vivo. Plkl may be a novel therapeutic target for gastric cancer.

关 键 词：胃肿瘤 RNA 细胞周期 保罗样激酶 裸鼠 细胞生长 

分 类 号：R735.2[医药卫生—肿瘤]