Tiam-1反义基因抗肿瘤侵袭转移作用的研究  

Effect of Tiam-1 Antisense Oligodeoxynucleotides (ASODNS)on Antimetasta-sis of Tumor

作　　者：李兆忠[1] 张玲[1] 毛海婷[1] 王芸[1] 李登华[1] 崔树龄[1] 

机构地区：[1]山东省医学科学院基础医学研究所,济南250062

出　　处：《中国肿瘤生物治疗杂志》2002年第4期269-271,共3页Chinese Journal of Cancer Biotherapy

基　　金：山东省自然科学基金(Y99C06)

摘　　要：目的:从基因—蛋白—细胞效应多层次水平,观察Tiam-1反义基因下调肿瘤细胞内转移相关基因Tiam-1 mRNA和调节细胞骨架结构的Rho蛋白表达水平,探讨遏制肿瘤侵袭转移的作用和机制。方法:采用反义核酸技术合成Tiam-1反义寡核苷酸,硫代化修饰与脂质体形成复合物(Tiam-1 ASODNS-Lf),转导入人高转移巨细胞肺腺癌PG细胞。采用RT-PCR技术检测Tiam-1的表达水平,流式细胞术检测Rho蛋白表达水平,Matrigel体外转移模型检测PG细胞的侵袭转移力。结果:Tiam-1 ASODNS-Lf抑制PG细胞Tiam-1 mRNA表达水平,其相对表达值从0.86下降为0.38;显著抑制PG细胞内Tiam-1介导的信号传递相关Rho蛋白表达水平,其表达率从27.96％降至16.86％(P<0.001)。显著抑制穿过人工基底膜的侵袭转移PC细胞数,从945±40下降为649±35(P<0.05)。结论:Tiam-1反义基因下调PG细胞中侵袭诱导基因mRNA表达及其介导的信号传递蛋白Rho的表达,从而遏制其侵袭和转移能力。Objective:Tiam-1 antisense gene was used to downregulate Tiam-1 mRNA and protein expression, thus tumor metastasis could be suppressed. Methods; In this study Tiam-1 mRNA was examined by RT-PCR. The expression of Rho protein was examined by flow cytometry. Basement membrane was used to examine the effect of Tiam-1 antisense gene on metastatic behavior of PG cells. Results; The relative expression level of Tiam-1 mRNA was reduced after exposure to Tiam-1 ASODNS-Lf. Only 72 hours of exposure to Tiam-1 ASODNS, cell metastasis of treated group was inhibitted significantly. The invasive cell numbers through matrigel reduced from 945±40 to 649 ±35 ( P < 0. 05 ). Tiam-1 ASODNS-Lf significantly inhibitted Rho protein expression ratio from 27. 86% to 16. 86% (P < 0. 001) , which related to signal transduction pathway. Conclusions: Antisense gene inhibits invasion and metastasis of tumor cells through a cascade reaction of gene-protein-cell effect and provides the theoretical and experimental bases for clinical therapy and study of antimetastasis.

关 键 词：反义寡核苷酸 Tiam-1基因 RHO蛋白 肿瘤转移 

分 类 号：R73－37[医药卫生—肿瘤]