Smad4基因对胰腺癌细胞E-cadherin、β-catenin表达及增殖的影响  

Effect of Gene Smad4 Transfection on Expression of E-cadherin and β-catenin and Proliferation in Human Pancreatic Cancer Cells

作　　者：刘庆宏[1] 于鸿[2] 黄俊星[3] 王朝夫[4] 施达仁[4] 

机构地区：[1]江苏省泰州市人民医院普外科,225300 [2]江苏省泰州市人民医院病理科,225300 [3]江苏省泰州市人民医院肿瘤科 [4]复旦大学附属肿瘤医院病理科,上海市200032

出　　处：《中华全科医学》2011年第12期1831-1832,1894,共3页Chinese Journal of General Practice

基　　金：江苏省医学重点人才基金资助项目(RC2007036);南通大学自然科学基金资助项目(10Z088)

摘　　要：目的探讨Smad4基因对人胰腺癌细胞BxPC3 E-cadherin、β-catenin表达及细胞增殖的影响,为试图运用Smad4对胰腺癌进行基因治疗提供实验依据。方法经脂质体介导将含有Smad4重组表达质粒转染人胰腺癌细胞Bx-PC3,用G418筛选及RT-PCR、Western blot鉴定;分别采用RT-PCR和Western blot检测转染阳性细胞克隆E-cadherin、β-catenin表达改变;又分别采用噻唑盐(MTT)比色法及5-溴-2-脱氧尿苷(BrdU)掺入法检测阳性细胞克隆的增殖活性。结果成功建立表达Smad4的阳性BxPC3克隆(S4-8,S4-23),并证实其E-cadherin、β-catenin mRNA及蛋白表达均明显升高。MTT法和BrdU掺入法显示,S4-8与S4-23细胞克隆增殖显著降低。结论 Smad4可增加胰腺癌细胞E-cadherin及β-catenin的表达并抑制其增殖,Smad4可能对胰腺癌具有基因治疗的效果。Objective To investigate the effect of Smad4 transfection on the proliferation and the expression of E-cadherin and β-catenin in human pancreatic cancer cell line BxPC-3 and to provide experimental foundation for Smad4 gene therapy for pancreatic cancer.Methods The Smad4 gene was transfected into BxPC-3 cells through lipofectin and positive clones were screened by G418.RT-PCR and Western blot were employed to identify mRNA and protein expressions of Smad4 in BxPC-3 cells,respectively.The expression of E-cadherin and β-catenin in positive clones was detected by RT-PCR and Western blot.Cell viability was assessed by MTT and 5＇-bromo-2＇-deoxyuridine（BrdU） incorporation assays.Results Positive clones S4-8 and S4-23 with Samd4 over-expression were successfully established.Compared with non-transfected control group,the expression of E-cadherin and β-catenin in S4-8 and S4-23 was increased significantly,and the proliferation level of S4-8 and S4-23 was decreased significantly.Conclusion Smad4 transfection can effectively enhance the expression of E-cadherin and β-catenin and inhibit proliferation in BxPC-3 cell which suggested a potential role for Smad4 in gene therapy of pancreatic cancer.

关 键 词：胰腺癌 SMAD4 上皮型钙黏蛋白 Β-连环蛋白 

分 类 号：R735.9[医药卫生—肿瘤] R73-3[医药卫生—临床医学]