MACC1调节肝星状细胞表达MMP-2、MMP-9对胃癌细胞迁移侵袭能力的影响

Effect of expressions of MMP-2 and MMP-9 regulated hepatic stellate cells by MACC1 on ability of migration and invasion of gastric carcinoma cells

目的探讨结肠癌相关转移基因(metastasis-associated in colon cancer-1,MACC1)对肝星状细胞(hepatic stellate cell,HSC)表达基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)和基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)的调节作用,及对胃癌细胞迁移和侵袭能力的影响。方法利用携带MACC1基因的重组慢病毒颗粒(LV-MACC1-GFP)、空载体慢病毒颗粒(LVGFP)感染HSC分别作为实验组、阴性对照组,未转染处理的HSC作为空白对照组,以上三组细胞按照常规培养,应用RT-PCR和Western blotting技术检测细胞中MMP-2、MMP-9 m RNA和蛋白表达。以上三组细胞分别与胃癌细胞MKN45体外共培养,Transwell实验观察三组不同培养条件下对胃癌细胞MKN45迁移侵袭能力的影响。结果与阴性对照组和空白对照组比较,实验组HSC MMP-2、MMP-9 m RNA和蛋白表达水平均明显上调(P<0.01)。Transwell实验中与实验组共培养的胃癌细胞MKN45迁移和侵袭能力较其他两组明显升高(P<0.01)。结论 MACC1可能通过调节HSC MMP-2、MMP-9的表达,促进胃癌细胞的迁移和侵袭。

Objective To investigate the regulatory effect of hepatic stellate cell （HSC） on expressions of MMP-2 and MMP-9 by MACC1 and effects on ability of migration and invasion of gastric carcinoma cells. Methods The recom- binant lentiviral particles of MACC1 （LV-MACC1-GFP） and lentiviral particles of empty vector （LV-GFP） were used to infect HSC, named overexpression group and vector group, non-transfeeted HSC was as blank control group. Cells in three groups were cultured in accordance with routine, the expressions of MMP-2 and MMP-9 in human HSC were detec- ted by Western blotting and RT-PCR. Cells in three groups were co-cultured with MKN45 gastric carcinoma cells in vitro, the capabilities of migration and invasion were measured by Transwell assay. Results Compared with the vector group and blank control group, the expressions of MMP-2 and MMP-9 mRNA and protein in overexpression group were significantly increased （P 〈 0.01 ）. Transwell test indicated that the capabilities of migration and invasion of MKN45 co- cultured with overexpression group were significantly higher than the other two groups （P 〈 0. 01 ）. Conclusion MACC1 may regulate the expressions of MMP-2 and MMP-9 in HSC, and enhance the ability of migration and invasion of gastric carcinoma cells.