组织因子途径抑制物2对胰腺癌细胞浸润能力的影响

Role of tissue factor pathway inhibitor-2 in pancreatic cancer cell invasion

目的:探讨TFPI-2表达与胰腺癌细胞体外和体内浸润能力之间的关系. 方法:脂质体介导人类TFPI-2真核表达载体pBos-Cite- neo/TFPI-2转染胰腺癌细胞系Panc-1,G418筛选阳性细胞,RT-PCR,Western blot技术检测转染前后胰腺癌细胞中TFPI-2 mRNA以及相应蛋白质的表达水平;利用Boyden小室检测转染前后胰腺癌细胞穿膜的细胞数,比较转染前后胰腺癌细胞的裸鼠成瘤大小,以此作为评价胰腺癌细胞体外和体内浸润能力的指标. 结果:转染成功的胰腺癌细胞证实有TFPI-2 mRNA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60.7±7.6,109.7±5.5和110.7±9.0,P<0.001);3 wk后裸鼠成瘤明显缩小(0.39 cm×0.36 cm,0.56 cm×0.50 cm和0.60 cm×0.52 cm,P<0.001),病理切片亦证实没有肌层浸润现象. 结论:TFPI-2表达可显著抑制胰腺癌细胞的体外和体内浸润能力,为进一步开展胰腺癌的基因治疗提供实验依据.

AIM: To elucidate the relation between TFPI-2 expression and pancreatic cancer cell invasion. METHODS: Human TFPI-2 expression vector pBos-Cite-neo/TFPI-2 was transfected into pancreatic cancer cell line Panc-1. After being selected by G418, transfected and nontransfected cells were screened for TFPI-2 mRNA and protein by reverse transcription-polymerase chain reaction and Western blot analysis, respectively. The number of transfected or nontransfected cells passing through membrane of Boyden chamber and the ability of tumori-genesis in nude mice was counted as the basis assessing invasive behaviors of the tumor cells. RESULTS: Expression of mRNA and protein of TFPI-2 was 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 non-expressing cells (60.7 ± 7.6,109.7 ± 5.5 or 110.7 ± 9.0, P<0.001); The size grown in nude mice was significantly smaller than its counterpart at 3 weeks (0.39 cm × 0.36 cm, 0.56 cm × 0.50 cm or 0.60 cm × 0.52 cm, P <0.001), which were also confirmed no apparent vertical invasion into the muscle layers by pathology. CONCLUSION: Expression of TFPI-2 may strongly inhibit the invasive ability of pancreatic tumor cells in vitro and in vivo, which provides an experimental basis for the treatment of human pancreatic cancer with gene-therapy.