mitofusin-2对乳腺癌细胞RECK基因表达及MMPs活性的影响

目的 探讨线粒体融合素基因-2(mitofusin-2)对乳腺癌MCF-7细胞株中RECK表达及MMP-9,MMP-2活性的影响.方法 利用脂质体lipofectamine2000将构建的重组真核表达质粒pEGFPmfn2转染MCF-7细胞.RT-PCR检测细胞mfn2和RECK基因mRNA的转录水平;Weastem Blot法检测mfn2及RECK蛋白的表达;明胶酶谱试验检测转染前后MMP-9及MMP-2的活性.结果 转染pEGFP-mfn2质粒的MCF-7细胞可以稳定高表达Mfn2;RECK基因在转染空质粒组和未转染组细胞中无表达;转染mfn2基因后RECK基因mRNA转录及蛋白的表达显著升高,且MMP-9及MMP-2的活性显著降-低(P＜0.05).结论 mfn2基因可激活MCF-7细胞中RECK基因的表达,并显著抑制其MMP-9及MMP2的活性.该途径可能是mfn2基因抗肿瘤作用的新机制.

Effect of mitofusin-2 gene on RECK expression and the activity of MMP-9 and MMP-2 in breast carcinoma cells

Objective:To investigate the regulative effect of the mfn2 gene on RECK expression and the activity of MMP-9 and MMP-2 in breast carcinoma cell (MCF-7 cell line). Methods :The pEGFP-mfn2 was transfected to MCF-7 cells with lipofectamine2000, the mRNA expression of mfn2 and RECK was assessed with RT-PCR, Western analysis was used to detect the protein expression of the mfn2 and RECK gene;and MMP-9 and MMP-2 activity was analyzed using gelatin zymograph. Results:After transfection of pEGFP-mfn2, the stable expression of Mitofusin-2 gene was detected in MCF-7 cells, the expression of RECK mRNA level and protein were increased significantly, while the MMP-9 and and MMP-2 activity level was markedly lower in the transfected Mitofusin-2 gene group than in the control(P<0.05). Conclusions:The mfn2 gene induces the expression of the RECK gene and inhibits the activity of MMP-9 and MMP-2 in MCF-7 cells, which might be a new mechanism of its anti-tumor effect.