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目的探讨组织因子(TF)在原发直肠癌中的表达意义,初步分析血管内皮生长因子(VEGF)对人直肠癌细胞(HT-29)TF表达的调控作用。方法(1)应用免疫组织化学染色法研究40例原发直肠癌组织、6例直肠良性腺瘤组织TF表达特点,分析其与转移和预后的关系;(2)以VEGF(25ng/ml)孵育HT-29细胞,分别应用免疫印记法、细胞免疫组织化学染色法和促凝血活性分析法检测不同时间点HT-29细胞TF的表达水平及活性。结果(1)40例原发直肠癌中20例(50%)TF表达阳性,正常黏膜和直肠良性腺瘤无TF表达,多因素回归分析表明,TF是影响原发直肠癌预后的因素之一(P=0.024);(2)直肠癌同时性和异时性肝转移组TF的表达均与肝转移显著相关(P=0.002和0.001);(3)Logistic回归分析表明,TF为直肠癌肝转移的影响因素(P=0.003);(4)静止状态下HT-29细胞存在TF基础表达,VEGF(25ng/ml)可诱导HT-29细胞TF表达增强,6h达高峰,刺激效应可维持24～32h。结论(1)TF可能参与原发直肠癌肝转移的生物学过程,其阳性表达可能作为高危患者术后肝转移的预测指标应用于临床;(2)TF是影响原发直肠癌预后的因素之一;(3)VEGF可能参与直肠癌细胞组织因子表达的上调过程。
Objective To investigate the expression of tissue factor (TF) in primary rectal cancer and to investigate the regulatory effect of vascular endothelial growth factor (VEGF) on TF expression in human rectal cancer cell line HT-29. Methods (1) The expression of TF in 40 cases of primary rectal cancer and 6 cases of rectal benign adenoma was studied by immunohistochemical staining. The relationship between metastasis and prognosis was analyzed. (2) Incubation with VEGF (25ng / ml) HT-29 cells were detected by Western blotting, immunohistochemical staining and procoagulant activity assay. The expression and activity of TF in HT-29 cells were detected at different time points. Results (1) TF expression was positive in 20 cases (50%) of 40 primary rectal cancers and no TF expression in normal mucosa and rectum benign adenoma. Multivariate regression analysis showed that TF was one of the factors affecting the prognosis of primary rectal cancer (P = 0.024). (2) The expression of TF in synchronous and heterotopic liver metastasis of rectal cancer patients was significantly correlated with liver metastasis (P = 0.002 and 0.001). (3) Logistic regression analysis showed that TF was (4) The basic TF expression was found in HT-29 cells at rest and VEGF (25ng / ml) increased the expression of TF in HT-29 cells, reaching the peak at 6h and the stimulatory effect could be maintained at 24 ~ 32h. Conclusions (1) TF may be involved in the biological process of primary hepatic metastasis of rectal cancer, and its positive expression may be used as a predictor of postoperative liver metastasis in high-risk patients. (2) TF is a factor affecting the prognosis of primary rectal cancer One; (3) VEGF may be involved in the up-regulation of tissue factor expression in rectal cancer cells.