摘要
背景与目的:组织因子途径抑制物2(tissuefactorpathwayinhibitor-2,TFPI-2)是一种新近发现的丝氨酸蛋白酶抑制物,可抑制包括纤溶酶、胰蛋白酶、基质金属蛋白酶在内的多种蛋白酶,而对尿激酶、组织型纤溶酶原激活剂以及凝血酶没有抑制作用。研究发现在多种肿瘤发生、发展过程中,TFPI-2常表达减少。本研究旨在探讨TFPI-2表达与卵巢癌细胞迁徙、浸润之间的关系。方法:脂质体介导人类TFPI-2真核表达载体pBos-Cite-neo/TFPI-2转染卵巢癌细胞株A2780,用G418筛选阳性细胞,经逆转录聚合酶链反应(reversetranscription-polymerasechainreaction,RT-PCR),Westernblot技术检测转染前后卵巢癌细胞中TFPI-2mRNA以及相应蛋白质表达水平;利用Boyden小室检测转染前后卵巢癌细胞穿膜的细胞数,此细胞数作为评价卵巢癌细胞迁徙、浸润能力的指标。结果:转染成功的卵巢癌细胞证实有TFPI-2mRNA和相应蛋白质的表达;与未转染的细胞相比,转染TFPI-2的细胞体外浸润能力显著下降(穿膜细胞数为59.3±6.5vs109.7±5.5,P<0.01);而迁徙能力无明显变化(穿膜细胞数为114.7±8.6vs127.3±7.1,P>0.05)。结论:TFPI-2表达可显著抑制卵巢癌细胞的浸润,为进一步开展肿瘤基因治疗提供实验依据。
BACKGROUND &OBJECTIVE: Human tissue factor pathway inhibitor 2 (TFPI 2) is a newly discovered serine protease inhibitor, which inhibits plasmin, trypsin, matrix metalloproteinase (MMPs), but not urokinase, tissue type plasminogen activators and thrombin. Earlier studies have shown that the production of TFPI 2 is downregulated during the progression of various cancers. The aim of this study was to elucidate the relationship between TFPI 2 expression and ovarian tumor migration and invasion. METHODS: Human TFPI 2 expression vector pBos Cite neo/TFPI 2 was transfected into ovarian tumor cells line A2780. After the transfected cells were screened by G418, transfected and nontransfected cells were examined for TFPI 2 mRNA and protein by reverse transcription polymerase chain reaction (RT PCR) and Western blot analysis, respectively. The number of transfected or nontransfected cells passing through membrane of Boyden chamber was counted as the basis assessing tumor cells migratory and invasive behaviors. RESULTS: Expression of mRNA and protein of TFPI 2 were confirmed in transfected cells. In invasion assay, the number of TFPI 2 expressing cells to traverse a Matrigel coated membrane was obviously decreased compared with that of nonexpressing cells (59.3±6.5 versus 109.7±5.5,P< 0.01); while in migration assay, no significant difference was observed between transfected and nontransfected cells (114.7±8.6 versus 127.3±7.1,P >0.05). CONCLUSION: Expression of TFPI 2 may strongly inhibit the invasive ability of ovarian tumor cells in vitro, but has no effect on the migratory ability, which provides an experimental basis for treating human ovarian tumor with gene therapy.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2003年第10期1038-1041,共4页
Chinese Journal of Cancer
