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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 cDNA transfection on the expression of matrix metalloproteinase (MMP) -7 and its inhibitor (TIMP) -1 in LoVo cells. Methods The LoVo cell line was transfected with pcDNA3.1 / Zeo vector constructed by antisense TF cDNA. The expression of ProMMP-7 in transfected and control cells was detected by Western blot and reverse transcription-polymerase chain reaction (RT-PCR) / TIMP-1 protein and mRNA expression levels. Results The expression of MMP-7 mRNA in LoVo cell line transfected with antisense TF cDNA was significantly lower in 10% fetal bovine serum-stimulated group than that in empty vector group and non-transfected cell group (0.07 ± 0.02, 0.25 ± 0.05,0.24 ± 0.06, P <0.05), and the expression of ProMMP-7 protein was also lower than the control group (0.10 ± 0.02,0.21 ± 0 .04,0.20 ± 0.03, P <0.05). There was no significant change in the expression of TIMP-1 in three groups of cells. The expression of MMP-7 was correlated with the expression of TF in each group (r = 0 .938, P <0.01). Conclusion The imbalance of MMP-7 / TIMP-1 expression plays a role in the invasion and metastasis of human colorectal cancer cells by tissue factor.