PTEN基因转染对人膀胱癌细胞系BIU87增殖及侵袭活性的影响  

Effects of PTEN Gene Transfection on Proliferation and Invasion of Human Bladder Cancer Cell Line BIU87

作　　者：张建华[1] 杨为民[1] 周四维[1] 陈忠[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院泌尿外科,湖北武汉430030

出　　处：《癌症》2006年第5期555-559,共5页Chinese Journal of Cancer

基　　金：武汉市科技攻关课题资助项目(No.20036004056)~~

摘　　要：背景与目的:PTEN(phosphataseandtensinhomologuedeletedfromchromosome10)是迄今为止发现的第一个具有磷酸酶活性的抑癌基因,在多种原发性恶性肿瘤和肿瘤细胞株中均存在有较高频率的缺失或突变,其失活与肿瘤的进程和预后相关。本实验研究外源性PTEN转染后,人膀胱癌细胞系BIU87增殖和侵袭活性的改变。方法:将携有PTEN基因的重组真核表达质粒pBp-PTEN转化大肠杆菌DH5α并扩增,抽提纯化质粒并进行酶切鉴定,pBp-PTEN体外转染BIU87细胞(pBp-PTEN-BIU87),筛选稳定转染的细胞并扩增培养,以转染了空质粒pBp的BIU87细胞(pBp-BIU87)和正常BIU87细胞为对照,用RT-PCR检测PTEN的表达情况,并用四甲基偶氮唑蓝比色法(MTT法)和细胞侵袭实验分别检测PTEN基因转染前、后BIU87细胞增殖和侵袭活性的变化。结果:pBp-PTEN的酶切鉴定证实含有目的基因PTEN;pBp-PTEN-BIU87细胞的RT-PCR产物经凝胶电泳有明显的阳性条带,而对照组则无;MTT发现,pBp-PTEN-BIU87细胞在第2、3和4天的吸光度(A值)明显低于两对照组(P<0.05),以正常BIU87细胞为对照,PTEN基因在第1、2、3、4天的细胞生长抑制率分别为4.27%、18.92%、19.54%、17.69%。细胞侵袭试验显示,各组细胞浸润穿透ECM膜的数目:pBp-PTEN-BIU8为39.3±7.7,BIU87和pBp-BIU87分别为48.1±13.2和48.9±11.0,pBp-PTEN-BIU87细胞明显少于两对照组(P<0.05)。结论:抑癌基因PTEN转染能降低膀胱癌细胞的增殖和侵袭活性。BACKGROUND ＆ OBJECTIVE： PTEN （phosphatase and tensin homologue deleted from chromosome 10） is the first antioncogene with phosphatase activity till now, and was found absent or mutational in many primary malignant tumors and cell lines. Its inactivation is correlated to tumor development and prognosis. This study was to investigate the effects of PTEN transfection on proliferation and invasion of human bladder cancer cell line BIU87. METHODS： A eukaryotic expression plasmid containing PTEN, pBp-PTEN, was introduced into E.coli DH5α and amplified. The plasmid was prepared and purified, and then identified by restrictive enzyme digestion. pBp-PTEN was transfected into BIU87 cells, and positive cell clones （pBp-PTEN-BIU87） were selected and amplified. Empty plasmid-transfected BIU87 cells （pBp-BIU87） and normal BIU87 cells were set as control. The expression of PTEN was detected by reverse transcription-polymerase chain reaction （RT-PCR）. Proliferation and invasion ability of BIU87 cells were measured before and after transfection by MTT assay and cell invasion assay. RESULTS： Plasmid pBp-PTEN containing PTEN was successfully constructed and transfected into BIU87 cells. After transfection, the inhibitory rates of cell growth at the first, second, third, and fourth days were 4.27%, 18.92%, 19.54%, and 17.69%, respectively. The penetrating cells were significantly less in pBp-PTEN-BIU87 group than in pBp-BIU87 and BIU87 groups （39.3± 7.7 vs. 48.1±13.2 and 48.9＋11.0, P〈0.05）. CONCLUSION： Transfection with PTEN might suppress proliferation and invasive ability of bladder cancer BIU87 cells.

关 键 词：PTEN基因 转染 膀胱肿瘤 BIU87细胞 增殖与侵袭 基因治疗 

分 类 号：R737.14[医药卫生—肿瘤]