组织因子途径抑制物2表达抑制卵巢癌细胞浸润能力的实验研究

Experimental study of tissue factor pathway inhibitor-2 expression inhibiting the invasive potential of human ovarian tumor cells

目的 本研究旨在探讨组织因子途径抑制物2(tissue factor pathway inhibitor-2,TFPI-2)表达对卵巢癌细胞体外、体内浸润能力的影响。方法 脂质体介导人类TFPI-2真核表达载体pBos-Cite-neo／TFPI-2转染卵巢癌细胞系A2780,用G418筛选阳性细胞,经逆转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR),Western blot技术检测转染前后卵巢癌细胞中TFPI-2 mRNA以及相应蛋白质表达情况;利用Boyden小室比较转染前后卵巢癌细胞穿膜的细胞数,通过接种BALB／c裸鼠,观察转染前后卵巢癌细胞在裸小鼠体内浸润转移范围的变化。结果 转染成功的卵巢癌细胞证实有TF-PI-2 mRNA和相应蛋白质的表达;转染TFPI-2的细胞体外浸润能力显著下降(59.3±6.5 vs 109.7±5.5,P<0.01),在裸小鼠体内浸润范围也明显缩小。结论TFPI-2表达可显著抑制卵巢癌细胞体外、体内的浸润能力,为进一步开展卵巢癌基因治疗提供实验依据。

Objective Human tissue factor pathway inhibitor-2 (TFPI-2) is a new serine protease inhibitor, which inhibits plas min, trypsin, matrix metalloproteinase (MMPs), rather than urokinase, tissue-type plasminogen activators and thrombin. Previous studies have shown that the production of TFPI-2 is downregulated during the invasion and metastasis of various cancers. The aim of this study was to explore the role of TFPI-2 expression in ovarian tumor invasion in vitro and in vivo. Methods Human TFPI-2 expression vector pBos-Cite-neo/TFPI-2 was transfected into ovarian tumor cells line A2780. After the transfected cells were selected by G418, transfected and nontransfected cells were screened for TFPI-2 mRNA and protein by reverse tran scription-polymerase chain reaction and western blot analysis, respectively. The number of transfected or nontransfected cells passing through membrane of Boyden chamber was counted as the basis assessing tumors cells invasive behaviors in vitro; the extent of invasion in vivo were evaluated by the subcutaneous injection of ovarian tumor cells to nude mice and followed by the pathological examination. Results Expression of mRNA and protein of TFPI-2 were confirmed in transfected cells. The number of TFPI-2-expiessing cells to traverse a Matrigel-coated membrane was obviously decreased compared with that of nonexpressing cells (59. 3±6. 5 vs 109. 7±5. 5, P<0. 01) ;and the extent of the invasion into the nude mice was significantly reduced. Conclusion TFPI-2 expression may obviously inhibit the invasion ability of ovarian tumor cells in vitro and in vivo , this provides an experimental basis for curing human ovarian tumor with gene-therapy.