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目的研究组织因子(TF)表达对人类大肠癌细胞(HT-29)侵袭能力的影响。方法(1)利用构建有正义/反义TFcDNA的质粒pcDNA3·1/Zeo分别转染HT-29细胞;(2)提取细胞的总蛋白,采用Westernblot检测TF的蛋白表达水平;(3)细胞分别进行体外基质胶(Matrigel)侵袭实验,观察细胞侵袭能力的变化;(4)细胞分别接种裸鼠,观察肿瘤生长,并检测肿瘤内的微血管密度。结果(1)与未转染的HT-29细胞相比,转染了正义和反义TFcDNA的HT-29细胞的TF蛋白表达水平以及细胞的体外侵袭能力分别升高和降低;(2)与未转染的HT-29细胞相比,转染了正义和反义TFcDNA的HT-29细胞在裸鼠体内侵袭能力以及肿瘤微血管密度分别升高和降低。结论组织因子的表达能够增强人类大肠癌HT-29细胞的体外和体内侵袭能力。
Objective To investigate the effect of tissue factor (TF) on invasion of human colorectal cancer cells (HT-29). Methods (1) HT-29 cells were transfected with pcDNA3.1 / Zeo plasmid containing sense / antisense TF cDNA respectively. (2) The total protein of cells was extracted and the protein expression of TF was detected by Western blot. (3) (4) The cells were inoculated with nude mice to observe the tumor growth and to detect the density of microvessels in the tumor. Results (1) Compared with untransfected HT-29 cells, TF protein expression of HT-29 cells transfected with sense and antisense TF cDNAs increased and decreased in vitro; (2) HT-29 cells transfected with sense and antisense TF cDNAs increased and decreased the invasiveness and tumor microvessel density in vivo in untransfected HT-29 cells, respectively. Conclusion Tissue factor expression enhances the invasiveness of human colorectal cancer HT-29 cells in vitro and in vivo.