组织因子表达与直肠癌肝转移的关系及其调控

Tissue factor expression in rectal cancer with liver metastasis and modulation of tissue factor expression in vitro

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 correlation of tissue factor (TF) expression between liver metastasis and prognosis in rectal cancer and study the modulation of tissue factor expression in HT 29 cells in vitro.Methods (1)TF expression of specimens from 40 rectal cancer and 6 benign rectal adenoma was studied by immunohistochemistry.Risk factors for hepatic metastasis and prognosis were analyzed with Logistic regression and multivariate statistics respectively. (2)HT 29 cells were incubated with VEGF (25 ng/ml).Western blot assay and cellular immunohistochemistry were applied to detect TF expression, and procoagulant activity assay for TF activity in different time course.Results (1) TF expression was positive in 20(50%) of 40 primary rectal cancer specimens, but negative in normal mucosa and rectal adenomas.(2)TF expression was a significant risk factor for hepatic metastasis (P=0 003) and prognosis (P=0 024).(3) TF expression was correlated to the occurrence of hepatic metastasis in both synchronous and metachronous hepatic metastasis group(P=0 002 and P=0 001 respectively).(4)Constitutive TF expression was observed on HT 29 cell in culture medium without serum.VEGF (25 ng/ml) rapidly increased TF expression of HT 29 cells, peaking at 6th hour of incubation and subsiding to normal in 24～32 hours Conclusions (1)TF expression is a possible indicator of both hepatic metastasis and prognosis for rectal cancer patients.(2)VEGF may be involved in the up regulation of TF expression in vitro