组织因子对肝癌细胞侵袭能力的影响

Effect of Tissue Factor on Invasive Ability of Hepatocellular Carcinoma

组织因子(tissue factor,TF)是机体外源性凝血途径的启动因子,发挥生理性止血的重要作用.近来研究表明,TF除凝血功能外尚与多种恶性肿瘤的血管生成,侵袭转移及预后密切相关.为了探讨TF对人类肝癌细胞的影响,将成功构建带有正义/反义TFcDNA的真核细胞表达质粒pcDNA3.1-TF(+)/(-)转染人肝癌细胞系HepG2,经药物筛选后获得稳定细胞克隆;应用RT-PCR和Western印迹检测内源性TF mRNA及蛋白质表达水平的变化;通过体外侵袭实验进一步分析对细胞侵袭能力所造成的影响.结果显示,转染pcDNA3.1-TF(+)质粒的细胞TF表达水平明显升高,相应的其侵袭能力明显增强,而转染pcDNA3.1-TF(-)质粒的细胞TF表达水平及体外侵袭能力显著下降.研究结果表明,TF可以增强人类肝癌细胞体外侵袭和转移能力,与肝癌的进展相关,可作为原发性肝癌治疗的一个新靶点进行研究.

Tissue factor （TF） is the initiator of the extrinsic blood coagulation cascade. The recent evidences indicate that TF is related to angiogenesis, invasion and prognosis of many malignant tumors. To explore the effect of TF on human hepatocareinoma, the eukaryotic expression plasmid pcDNA3.1-TF （ ＋ ）/（ - ） was transfected into human hepatocellular carcinoma cell line HepG2. After 4-6 weeks selected by Zeocin, the stable expression of sense or antisense TF cDNA cell clone was obtained. TF expression in HepG2 was determined by RT-PCR and Western blotting. Matrigel invasion assay was used to analysis the invasive ability of HepG2 cells in vitro. The results indicated that there was an elevated expression of TF in the cells transfected with pcDNA3.1-TF（ ＋ ） as compared with the cells without transfection, furthermore, the invasive ability of these cells increased. Whereas in HepG2 cells with pcDNA3.1-TF （ - ） transfected, both the expression of TF and the invasive ability were decreased. These results suggest that TF can enhance the invasion and metastasis ability of tumor cell in vitro. TF might play an important role in the progression of human hepatocellular carcinoma and it can be researched as a new therapeutic target of hepatocarcinoma in the future.