MT1-MMP反义核酸抑制高转移人卵巢癌SW626细胞的侵袭效应  

Inhibitory Effect of MT1-MMP Antisense Nucleotide on Invasion of Human Highly Metastatic Ovarian Cancer Cell Line SW626

作　　者：吴明富[1] 廖国宁[1] 贾平[1] 奚玲[1] 卢运萍[1] 马丁[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院妇产科,湖北武汉430030

出　　处：《癌症》2004年第11期1263-1266,共4页Chinese Journal of Cancer

基　　金：国家杰出青年基金资助项目(No.30025017);国家重点基础研究发展规划项目(No.2002CB513107)~~

摘　　要：背景与目的:MT1-MMP(MMP-14)是新发现的一种膜型基质金属蛋白酶,研究表明MT1-MMP在肿瘤转移中发挥关键性作用。本研究应用反义核酸技术,观察了MT1-MMP反义核酸对高转移人卵巢癌SW626细胞体外生长和侵袭的抑制效应。方法:应用自行设计的引物,借助RT-PCR技术获得两端含不同限制酶位点的MT1-MMPcDNA片段,反向插入pcDNA3.1中,并应用脂质体介导的基因转染技术,将重组质粒导入SW626细胞中,应用MTT、Westernblot、明胶酶谱和体外侵袭实验等方法观察了SW626细胞转染前后,细胞生长、MT1-MMP蛋白表达、MMP-2和MMP-9酶活性及细胞体外侵袭能力等指标的变化。结果:成功构建了反义MT1-MMP真核表达载体(pMMP14as),将其转染入SW626细胞后,与空质粒转染组相比,MT1-MMP反义核酸转染组细胞MT1-MMP表达显著降低,抑制率为65.8%;MMP-2酶原的活化受到明显抑制;pMMP14as转染组的穿膜细胞百分数为(63.3±5.8)%,明显低于空质粒转染组(97.6±7.5)(P<0.05)%。结论:MT1-MMP反义核酸可明显抑制高转移SW626细胞体外生长和侵袭效应,提示MT1-MMP可作为人卵巢癌抗侵袭治疗的分子靶点。BACKGROUND &OBJECTIVE: Membrane type 1 matrix metalloproteinase (MT1 MMP/MMP 14) is a newly discovered enzyme, which plays a key role in tumor metastasis. This study was to observe inhibitory effect of MT1 MMP antisense nucleotide on proliferation and invasive potential of human highly metastatic ovarian carcinoma cell line SW626. METHODS: Reverse transcriptase polymerase chain reaction (RT PCR) was used to amplify MT1 MMP cDNA fragments with 2 different restriction sites at its 5' end. RT PCR products were cloned into plasmid pcDNA3.1 in antisense direction. The recombinant pMMP14as was transfected into SW626 cells. Changes of cell proliferation, MT1 MMP protein expression, activities of MMP 2 and MMP 9, and cell invasion ability were detected by MTT assay, Western blot, optimized gelatin zymography, and matrigel in vitro invasion assay, respectively. RESULTS: Antisense MT1 MMP eukaryotic expression vector pMMP14as was constructed successfully. After 48 h transfection with pMMP14as, proliferation of pMMP14as transfected SW626 cells was significantly lower than that of control cells. Compared with control cells, the expression of endogenous MT1 MMP protein in pMMP14as transfected cells was decreased with a inhibition rate of 65.8%. The activation of proMMP 2 was remarkably inhibited, and the mean invasive cell percentage was (63.3±5.8)%in pMMP14as transfected cells, which was far less than (97.6±7.5)%in control cells (P< 0.05). CONCLUSION: Both cell proliferation and invasive potential of SW626 cells wereinhibited effectively by antisense MT1 MMP, suggesting that MT1 MMP may be a proper molecular target of anti invasion therapy for human ovarian cancer.

关 键 词：MT1-MMP 卵巢肿瘤 侵袭转移 

分 类 号：R737.31[医药卫生—肿瘤]