组织因子/活化因子Ⅶ复合物影响人卵巢癌细胞侵袭及转移能力的研究

目的　研究组织因子 /活化因子Ⅶ (TF/FⅦa)复合物对人卵巢癌细胞体外侵袭能力及体内转移能力的影响。方法　①采用分子克隆及基因转染技术构建高效表达TF的人卵巢癌细胞系A2 780 /TF ;②采用Boyden小室法计数FⅦa刺激后A2 780、A2 780 /TF穿透Matrigel迁移到PVPF膜背面的细胞数 ;③建立人卵巢癌细胞转移的实验动物模型 ,研究TF对人卵巢癌细胞在裸鼠体内形成转移灶的能力。结果　①经分子克隆技术构建得到pcDNA3 TFcDNA重组体 ;②稳定转染的A2 780 /TF细胞内TFmRNA水平显著增高 :转染细胞为 3.99± 0 .15 ,未转染细胞为 0 .97± 0 .2 3(P <0 .0 1) ;转染细胞表面TF表达也显著增高 :转染细胞中约 (48.5 6± 9.5 3) %细胞表面有TF抗原 ,而未转染细胞中仅 (2 .73± 1.15 ) % (P <0 .0 1) ;③A2 780 /TF细胞穿膜细胞数为 15 7.3± 19.2 ,经FⅦa刺激后穿膜细胞数显著增多 ,为 4 4 7.7± 39.4 (P <0 .0 1) ;与FⅦa和抗TF单抗共孵育的A2 780 /TF ,穿膜细胞数接近刺激前的基础水平 ;④种植A2 780细胞的裸鼠中 2 2 .2 %可见肺部转移灶 ,种植A2 780 /TF细胞的裸鼠中 88.9%发生肺转移 (P <0 .0 1)。结论　①将所构建的TF真核表达载体转入人卵巢癌细胞系 ,获得稳定、高效表达TF的人卵巢癌细胞系A2 780 /TF ,为研究TF

The effects of tissue factor/activated factor Ⅶ complex on the invasion and metastasis of human ovarian cancer

OBJECTIVE: To explore the role of tissue factor/activated factor VII (TF/FVIIa) complex in human ovarian cancer invasion and metastasis. METHODS: (1) Constructed an expression vector of TF, pcDNA3-TF and established a human ovarian cell line A2780/TF expressing high level TF by using molecular cloning and gene transfection techniques. (2) By Boyden chamber assay to count the numbers of A2780 and A2780/TF cells that penetrated the matrigel to the back of PVPF membrane after FVIIa stimulation. (3) BALB/c nude mice were used to establish experimental model of metastasis with A2780 or A2780/TF and the lung tissue sections were examined by microscopy for cancer metastasis. RESULTS: (1) Compared with their parental A2780 cells, A2780/TF cells expressed high level of TF mRNA (3.99 +/- 0.15 vs 0.97 +/- 0.23, P < 0.01) and TF antigen on cell surface [(48.56 +/- 9.53)% vs (2.73 +/- 1.15)%, P < 0.01]. (2) After stimulation, the A2780/TF cell number on the back of PVPF membrane increased from basal level 157.3 +/- 19.2 to 447.7 +/- 39.4 (P < 0.01), which could decreased to basal level when coincubated with anti-TF antibody. (3) Cancer metastasis was found in 22.2% of nude mice transplanted with A2780 cells, while in 88.9% of those transplanted with A2780/TF cells. CONCLUSION: TF could promote the invasion and metastasis of human ovarian cancer cells through TF/FVIIa pathway.