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目的:探讨组织因子促进人类大肠癌细胞侵袭的机制,研究其与MMP 2和MMP 9的相关性。方法:获取转染正义/反义TFcDNA的人类大肠癌HT 29细胞系/LoVo细胞系裸鼠皮下种植瘤标本,应用Westernblot和RT PCR技术,检测转染组和对照组肿瘤组织中MMP 2和MMP 9的蛋白和mRNA表达水平;同时采用明胶酶谱法检测转染反义LoVo细胞系培养上清中的MMP 2和MMP 9活性的变化。结果:与对照组相比较,转染正义TFcDNA的HT 29细胞系成瘤标本TF表达升高,其所对应的MMP 9和MMP 2蛋白水平和mRNA水平表达也明显升高; 转染反义TFcDNA的LoVo细胞系成瘤标本中的TF,MMP 9和MMP 2蛋白和mRNA水平表达明显低于对照组,转染反义TFcDNA的LoVo细胞系培养上清液中的MMP 9和MMP 2活性较对照组低;MMP 9和MMP 2表达与组织因子表达具有相关性。结论:组织因子促进人类大肠癌细胞的侵袭,其部分机制可能与组织因子上调MMP 9和MMP 2的表达有关,组织因子可能是大肠癌细胞分泌MMPs的内在激动剂。
OBJECTIVE: To investigate the mechanism of tissue factor in promoting the invasion of human colorectal cancer cells, and to study its correlation with MMP-2 and MMP-9. Methods: Human colorectal cancer HT29 cell line / LoVo cell line transfected with sense / antisense TF cDNAs were subcutaneously implanted into nude mice. Western blot and RT-PCR were used to detect the expression of MMP-2 and MMP in tumor tissues of transfected and control groups 9 protein and mRNA expression level; while gelatin zymography assay transfected antisense LoVo cell line supernatant of MMP 2 and MMP 9 activity changes. Results: Compared with the control group, the expression of TF in tumor-bearing samples of HT29 cell line transfected with sense TFcDNA increased, and the corresponding MMP9 and MMP2 protein levels and mRNA levels were also significantly increased; antisense TF cDNA LoVo cell line tumor-bearing specimens of TF, MMP 9 and MMP 2 protein and mRNA levels were significantly lower than the control group transfected antisense TF cDNADNA LoVo cell line culture supernatant MMP 9 and MMP 2 activity than the control Group low; MMP 9 and MMP 2 expression and tissue factor expression has relevance. CONCLUSION: Tissue factor can promote the invasion of human colorectal cancer cells. Its mechanism may be related to up-regulation of MMP-9 and MMP-2 expression by tissue factor. Tissue factor may be an intrinsic agonist of MMPs secretion by colorectal cancer cells.