RNAi沉默Polo-like kinase-1基因表达对大肠癌细胞端粒酶活性的影响  

Effects of RNA interference polo-like kinase-1 gene on telomerase activity of human colon cancer cell

作　　者：倪国华[1] 范钰[2] 陈坚[3] 钱立平[3] 林庚金[3] 陈功星[4] 丁佳逸[4] 郑树[4] 

机构地区：[1]江苏省丹阳市人民医院肿瘤科,江苏丹阳212300 [2]江苏大学附属人民医院肿瘤研究所化疗科,江苏镇江212002 [3]复旦大学附属华山医院消化科,上海200040 [4]浙江大学肿瘤研究所,浙江杭州310009

出　　处：《复旦学报（医学版）》2009年第1期14-18,共5页Fudan University Journal of Medical Sciences

基　　金：镇江市科技计划基金项目(SH2006019);中国博士后科学基金项目(2003033547)

摘　　要：目的探讨polo-like kinase-1(PLK1)基因对大肠癌细胞增殖和端粒酶活性的影响。方法根据PLK1基因序列特点,设计并用化学方法合成小干扰核糖核酸分子(small interfering RNA,si RNA),转染人大肠癌SW480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。分别采用MTT法和TRAP-ELISA方法检测PLK1基因转染对大肠癌细胞增殖和端粒酶活性的影响。结果所设计的5个si RNA均能明显抑制大肠癌SW480细胞PLK1 mRNA水平,以P4效果最好。以P4转染处理大肠癌细胞后,PLK1 mRNA水平和蛋白水平明显下调,且呈浓度和时间依赖性。MTT和TRAP-ELISA方法检测发现,P4si RNA转染组细胞增殖和端粒酶活性明显受到抑制,且呈浓度和时间依赖性(P<0.05,P<0.05)。结论PLK1基因对大肠癌细胞增殖具有重要的调控作用;以PLK1 si RNA转染处理大肠癌细胞,可明显抑制大肠癌细胞的恶性增殖,其机制可能与抑制端粒酶活性有关。Objective To investigate the role of polo-like kinase-1 （PLK1） on proliferation and telomerase activity of human colon cancer cells. Methods After SW480 colon cancer cell were transfected with small interfering RNA （siRNA） against PLK1, the real time RT-PCR and Western blot assay were used to examine PLK1 gene expression in all cancer cells. The proliferation and telomerase activity of cancer cells were determined by MTT and telomeric repeat amplification protocol enzyme-linked immunoadsordent assay （TRAP-ELISA） ,respectively. Results Expression of PLK1 of SW480 cancer cell transfected with siRNA down-regulated significantly. Transfection of PLK1 siRNA resulted significantly in inhibition of colon cancer cell in vitro. The results from TRAP showed that cancer ceils exhibited marked apoptosis, in time-and dose- dependent pattern. Conclusions RNA interference PLK1 could inhibit proliferation through inducing suppressing telomerase activity of human colon cancer cell.

关 键 词：大肠肿瘤 polo—like kinase-1 RNA干扰 小干扰RNA 端粒酶 

分 类 号：R735.34[医药卫生—肿瘤]