Pin1siRNA对宫颈癌细胞SiHa增殖和凋亡的影响Impact of Pin1 inhibition by siRNA on proliferation and apoptosis in human cervical cancer cell line SiHa
钱颖;贺晓琪;蔡惠兰;董卫红;王泽华;
摘要(Abstract)：
目的:通过小干扰RNA(siRNA)载体介导抑制肽基脯氨酰同分异构酶(Pin1)表达,探讨Pin1对宫颈癌细胞SiHa增殖与凋亡的影响。方法:构建pGPU6/GFP/Neo-Pin1 siRNA表达载体,脂质体介导将其转染至宫颈癌细胞SiHa中;实验设3组:实验组(SiHa/pGPU6-Pin1 siRNA组)、阴性对照组(SiHa/pGPU6-con组)和空白对照组(SiHa)。采用RT-PCR、Western blot法检测Pin1的表达;MTT法检测细胞增殖活性;原位末端标记法(TUNEL)和流式细胞术(FCM)检测细胞凋亡。结果:转染SiHa/pGPU6-Pin1siRNA的SiHa细胞中,Pin1 mRNA和蛋白表达分别下调68.1%和54.8%,细胞增殖显著抑制。TUNEL显示典型细胞凋亡特征,SiHa/pGPU6-Pin1 siRNA组的凋亡指数(AI)为(29.6±14.58)%,显著高于SiHa/pGPU6-con组[(8.44±5.65)%]和空白对照组[(5.34±4.47)%](P<0.05)。SiHa/pGPU6-Pin1 siRNA组的细胞凋亡率为(30.95±16.47)%,显著高于SiHa/pGPU6-con组[(6.18±6.10)%]和空白对照组[(5.95±4.99)%](P<0.05)。结论:RNA干扰技术能特异高效的抑制Pin1基因的表达,Pin1基因的下调能抑制宫颈癌细胞增殖并促进细胞凋亡,提示Pin1可能成为治疗宫颈癌的新靶点。
关键词(KeyWords)： RNA干扰;肽基脯氨酰顺反异构酶;宫颈癌细胞;增殖;凋亡
基金项目(Foundation): 国家自然科学基金资助项目(No:81072134);; 上海市浦东新区卫生系统优秀青年医学人才培养计划(No:PWRq2010-05);; 同济大学青年优秀人才培养计划(No:2011KJ008)
作者(Authors): 钱颖;贺晓琪;蔡惠兰;董卫红;王泽华;
DOI: 10.13283/j.cnki.xdfckjz.2013.10.006
参考文献(References)：
- [1]Theuerkorn M,Fischer G,Schiene-Fischer C.Prolyl cis/trans isomerase signalling pathways in cancer[J].Curr Opin Pharmacol,2011,11(4):281-287
- [2]Liou YC,Zhou XZ,Lu KP.Prolyl isomerase Pin1 as a molecular switch to determine the fate of phosphoproteins[J].Trends Biochem Sci,2011,36(10):501-514
- [3]Lee TH,Pastorino L,Lu KP.Peptidyl-prolyl cis-trans isomerase Pin1 in ageing,cancer and Alzheimer disease[J].Expert Rev Mol Med,2011,13:e21
- [4]Lu KP,Suizu F,Zhou XZ,et al.Targeting carcinogenesis:a role for the prolyl isomerase Pin1?Mol Carcinog[J].2006,45(6):397-402
- [5]Aagaard L,Rossi JJ.RNAi therapeutics:principles,prospects and challenges[J].Adv Drug Deliv Rev,2007,59(2-3):75-86
- [6]Lu KP,Hanes SD,Hunter T.A human peptidyl-prolyl isomerase essential for regulation of mitosis[J].Nature,1996,380(6574):544-547
- [7]Ryo A,Liou YC,Lu KP,et al.Prolyl isomerase Pin1:a catalyst for oncogenesis and a potential therapeutic target in cancer[J].J Cell Sci,2003,116(Pt 5):773-783
- [8]Bao L,Kimzey A,Sauter G,Prevalent overexpression of prolyl isomerase Pin1 in human cancers[J].Am J Pathol,2004,May;164(5):1727-1737
- [9]Ayala G,Wang D,Wulf G,et al.The prolyl isomerase Pin1 is a novel prognostic marker in human prostate cancer[J].Cancer Res,2003,63(19):6244-6251
- [10]Ryo A,Uemura H,Ishiguro H,et al.Stable suppression of tumorigenicity by Pin1-targeted RNA interference in prostate cancer[J].Clin Cancer Res,2005,11(20):7523-7531
- [11]Lu KP,Finn G,Lee TH,et al.Prolyl cis-trans isomerization as a molecular timer[J].Nat Chem Biol,2007,3(10):619-629
- [12]Lu KP,Zhou XZ.The prolyl isomerase PIN1:a pivotal new twist in phosphorylation signalling and disease[J].Nat Rev Mol Cell Biol,2007,8(11):904-916