组织因子／活性凝血7因子体外激活大肠癌LOVO细胞基质金属蛋白酶-7基因启动子  

TF/FViia complex activates the promoter of MMP-7 in LOVO cells of colon cancer in vitro

作　　者：张剑权[1] 万远廉[1] 刘玉村[1] 汪欣[1] 汤坚强[1] 吴涛[1] 朱静[1] 潘义生[1] 

机构地区：[1]北京大学第一医院普通外科,100034

出　　处：《中华实验外科杂志》2008年第3期279-281,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30471683）

摘　　要：目的观察组织因子（TF）／活性凝血7因子（FVIIa）复合物对大肠癌LOVO细胞系基质金属蛋白酶-7（MMP-7）基因启动子的激活。方法靶向组织因子基因的小干扰RNA（siRNA）构建及稳定转染获得TFRNAi LOVO细胞系；Western blot检测组织因子的干扰效率；NNP-7基因上游5′非翻译区DNA调节序列及缺失体克隆到pGL3-basic荧光素酶报告基因载体；双报告基因检测FVIIa因子存在时，MMfr7．luc1592、MMP7．luc978、MMP7．luc615瞬时转染LOVO细胞或TFRAi LOVO细胞启动子活性。结果Western blot灰度半定量显示TFRNAi LOVO细胞中组织因子蛋白表达水平较LOVO细胞下降41．7％；双报告基因检测FVIIa因子存在时MMP7．luc1592具有启动子活性并依赖组织因子，活性区域位于MMP-7基因转录起始点上游-978bp及-615bp之间。结论TF／FVIIa能直接激活大肠癌LOVO细胞系MMP-7基因启动子。Objective To investigate activation of the promoter of MMP-7 in LOVO cells of colon cancer induced by TF/FVIIa complex. Methods LOVO cells were transfeeted stably with RNAi plasmid targeting to tissue factor to get TFRNAi LOVO cells and the efficiency of interfering in TFRNAi LOVO cells was detected by using Western blot. The 5′ UTR of human MMP-7 gene and deletion constructed were cloned into the pGL3-basic luciferase reporter vector. Promotor activity of LOVO cells or TFRNAi LOVO cells transfected with MMP7. luc1592, MMP7. luc 978 or MMP7. luc 615 transiently exposured to FVIIa were analyzed by the Dual-Lucfferase Reporter Assay System. Results The protein expression level of TF gene was reduced by 41.7% in TFRNAi LOVO cells as compared with that in LOVO ceils. MMP7. luc1592 contained promoter activity and was dependent on TF in the presence of FVIIa, and its active region was located between -978bp and -615bp upstream of the transcription initiation site of the published MMP-7 gene. Conclusion TF/FVIIa complex can activate the promoter of MMP-7 in LOVO cells of colon cancer directly.

关 键 词：结肠癌 组织因子 基质金属蛋白酶-7 启动子 

分 类 号：R735.34[医药卫生—肿瘤] R735.2[医药卫生—临床医学]