DAPK1基因转染对高转移性肺癌细胞PGCl_3的影响  

Effect of DAPK1 Gene Transfection on High metastasis Non-small Lung Cancer Cell PGCl_3

作　　者：张海涛[1] 朱振宇[1] 冯哲玲[1] 李秀英[1] 李民友[1] 马涧泉[1] 梁念慈[2] 

机构地区：[1]中山大学中山医学院生化教研室,广东广州510089 [2]广东医学院生化教研室,广东湛江524023

出　　处：《癌症》2004年第5期497-501,共5页Chinese Journal of Cancer

基　　金：国家教委留学回国人员启动基金资助项目犤2000犦(No.479)~~

摘　　要：背景与目的:死亡相关蛋白激酶DAPK1失活是影响非小细胞肺癌患者生存的独立因子,过表达的DAPK1可以诱导肿瘤细胞凋亡,抑制肿瘤转移,但抑制转移的机制尚未完全清楚。本文探讨DAPK1基因抑制高转移性肺癌PGCl3细胞生长及转移的可能机制。方法:利用基因重组技术构建含DAPK1基因开放读码框(ORF)的真核表达载体pcDNA3.1-DAPK1,用脂质体LipofectAMINE2000介导转染PGCl3细胞系,检测转染后PGCl3细胞的生长曲线、体外软琼脂克隆形成率、体外侵袭、运动和粘附能力的变化,同时检测了胶原酶活性,p53,bcl-2基因表达的变化。结果:转染DAPK1基因的细胞生长比空白组及pcDNA3.1转染组减缓;体外软琼脂克隆形成率下降P<0.05;pcDNA3.1-DAPK1转染组体外侵袭能力是空白组的68.5%,而pcDNA3.1转染组是空白组的88.0%;pcDNA3.1-DAPK1转染组运动能力是空白组的87.3%,pcDNA3.1转染组是空白组的95.7%;pcDNA3.1-DAPK1转染组粘附能力是空白组的62.7%,pcDNA3.1转染组是空白组的91.2%;各组的胶原酶变化不明显;pcDNA3.1-DAPK1转染组p53基因表达升高,bcl-2基因下调。结论:DAPK1基因的过表达可以在一定程度上逆转PGCl3细胞的恶性表型,使PGCl3细胞生长受抑制,体外侵袭、运动和粘附能力下降,p53基因表达的上调及bcl-2基因表达下调。BACKGROUND &OBJECTIVE: Loss of activity of death associated protein kinase 1 (DAPK1) may be an independent factor affecting survival of non small cell lung cancer patients. DAPK1 over expression can induce cell apoptosis and inhibit tumor cell metastasis. However, the mechanism of DAPK1 inhibiting metastasis was unclear yet. This study was designed to investigate the mechanism of DAPK1 affecting PGCl3 cellsgrowth and metastasis. METHODS: Open read frame (ORF) of DAPK1 gene was recombined into eukaryotic express vector pcDNA3.1. The PGCl3 cell line, a high metastasis lung tumor cell line, was transfected with pcDNA3.1 DAPK1 by lipofectamine2000. The growth curve of PGCl3 cells, colony formation assays, invasive, migration and adhesion ability were evaluated. Gelatinase secretion of PGCl3 cells treated with DAPK1 and expression of p53 and bcl 2 gene in PGCl3 cells were determined. RESULTS: PGCl3 cells of pcDNA3.1 DAPK1 transfected group grew slower than blank group and pcDNA3.1 transfected group. The numbers of colony formation of pcDNA3.1 DAPK1 transfected group reduced in comparison with blank group and pcDNA3.1 transfected group (P< 0.05). Invasive ability of pcDNA3.1 DAPK1 transfected group was 68.5%to blank group, and pcDNA3.1 transfected group was 88%to blank group. Migration ability of pcDNA3.1 DAPK1 transfected group was 87.3%to blank group and pcDNA3.1 transfected group was 95.7%to blank group. Adhesion ability of pcDNA3.1 DAPK1 transfected group was 62.7%to blank group, and pcDNA3.1 transfected group was 91.2%to blank group. Changes in PGCl3 cells secreting gelatinase have not been observed in different groups. Expression of p53 gene was upregulated in pcDNA3.1 DAPK1 transfected group, while expression of bcl 2 gene was downregulated. CONCLUSION: DAPK1 gene over expression could suppress PGCl3 cells malignant phenotype, inhibit PGCl3 cells growth, invasive, migration and adhesion ability, upregulate p53 gene and downregulate bcl 2 gene. All of these changes may be the mechanism

关 键 词：DAPKl基因转染 高转移性肺癌细胞 PGCL3 影响因素 

分 类 号：R734.2[医药卫生—肿瘤]