反义组织因子cDNA转染对LoVo细胞基质金属蛋白酶-7及其抑制物-1表达的影响

Effect of antisense tissue factor cDNA transfection on the MMP-7/TIMP-1 expression in LoVo cells

目的探讨反义组织因子cDNA转染对LoVo细胞基质金属蛋白酶(MMP)-7及其抑制物(TIMP)-1表达的影响。方法利用构建有反义TFcDNA的质粒pcDNA3．1/Zeo转染LoVo细胞系,应用Western blot和逆转录-聚合酶链反应(RT-PCR)方法检测转染组和对照组细胞系中ProMMP-7/TIMP-1蛋白和mRNA表达水平。结果10%胎牛血清刺激组中,转染反义TFcDNA的LoVo细胞系中MMP-7 mRNA的表达水平明显低于空载体组和未转染细胞组(分别为0．07±0．02、0．25±0．05、0．24±0．06,P＜0．05),其ProMMP-7蛋白表达也低于对照组(分别为0．10±0．02、0．21±0．04、0．20±0．03,P＜0．05)；TIMP-1在三组细胞中的表达未见明显改变；MMP-7表达与各组细胞中的TF表达相关(r=0．938,P＜0．01)。结论MMP-7/TIMP-1表达的失平衡在组织因子促进人类大肠癌细胞侵袭转移的过程中发挥作用。

Objective To investigate the effect of antisense tissue factor expression cDNA transfection on the MMP-7/TIMP-1 expression in LoVo cells. Methods The eurokaryotic expression vectors pcDNA3.1/Zeo bearing antisense TFcDNA was transfected into LoVo cells. The expression of MMP-7 and TIMP-1 at the protein and mRNA levels was detected by Western Blot and RT-PCR respectively. Result The expression of ProMMP-7 at the protein and mRNA level in the group of LoVo cells with antisense-TFcDNA was significantly decreased as compared with that in pcDNA3.1/Zeo and untransfected LoVo cells. Relative expression of MMP-7 mRNA was 0.07 ± 0.02,0.25 ± 0.05 and 0.24 ± 0.06 respectively, and that of ProMMP-7 protein was 0.10 ± 0.02,0.21 ± 0.04 and 0.20 ± 0.03 respectively. The expression of TIMP-1 was not altered. The expression of MMP-7 was closely correlated to the ex- pression of tissue factor in LoVo cells （ r = 0. 938, P 〈 0.01 ）. Conclusion The imbalance of expression between MMP-7 and TIMP-1 plays a partial role in the mechanism by which tissue factor promotes the invasion and metastasis of human colon cancer.