组织因子促进人类大肠癌细胞的侵袭及与基质金属蛋白酶类的相关性研究  

Role of tissue factor in the invasion of cultured human colon carcinoma cells and its correlation with Matrix metalloproteinases

作　　者：汤坚强[1] 万远廉[1] 刘玉村[1] 戎龙[1] 汪欣[1] 吴涛[1] 潘义生[1] 叶京明[1] 姚宏伟[2] 朱静[1] 

机构地区：[1]北京大学第一医院普通外科,北京100034 [2]北京大学第三医院普通外科

出　　处：《北京大学学报（医学版）》2005年第3期265-268,共4页Journal of Peking University:Health Sciences

基　　金：国家自然科学基金(30471683);教育部科学技术研究重点项目资助(02003 )~~

摘　　要：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 role of tissue factor expression in the invasive ability of human colon carcinoma cells and to analyze the correlation between tissue factor and MMP-2 and MMP-9. Methods: HT-29 cells with sense-TFcDNA transfection and LoVo cells with antisense-TFcDNA transfection were implanted subcutaneously into nude mice as well as controls. The expressions of MMP-9 and MMP-2 at the protein and mRNA levels were detected by Western blot and RT-PCR respectively; Gelatin zymography was used for assay of the MMP-9 and MMP-2 activiies in the supernatant cultured media of LoVo cells with antisense-TFcDNA transfection and controls. Results: The expressions of MMP-9 and MMP-2 at the protein and mRNA levels in the group of HT-29 cells with sense-TFcDNA were significantly increased with untransfected HT-29 cells.The expression in the group of LoVo cells with antisense- TFcDNA had a significant decrease than that of the controls.The MMP-9 and MMP-2 activities of LoVo cells with antisense-TFcDNA were weakened because of the lower expression of tissue factor by gelatin zymography. The expression of MMP-9 and MMP-2 were closely correlated to the expression of tissue factor. Conclusion: Tissue factor can increase the invasive ability of human colon carcinoma cells. The partial mechanism is that TF can upregulate the expressions of MMP-9 and MMP-2.Tissue factor may be an inner agonist in the secrection of MMPs in colon carcinoma cells.

关 键 词：组织因子 大肠癌细胞 基质金属蛋白酶类 相关性研究 人类 HT-29细胞系 mRNA水平 Western MMP-9活性 MMP-2蛋白 MMP-2表达 LoVo mRNA表达 转染反义 癌细胞侵袭 皮下种植瘤 PCR技术 明胶酶谱法 培养上清液 对照组 blot 肿瘤组织 

分 类 号：R541.4[医药卫生—心血管疾病] R735.34[医药卫生—内科学]