转化生长因子β1/细胞外信号调节激酶/基质金属蛋白酶通路对腺样囊性癌侵袭和迁移的影响

Experiment study on the role and mechanism of TGF-β1/ERK1/2/MMP-2 signaling pathways in the migration and invasion process of ACC

目的探讨转化生长因子β1/细胞外信号调节激酶/基质金属蛋白酶2(TGF-β1/ERK1/2/MMP-2)通路在腺样囊性癌(ACC)侵袭和迁移过程中的作用及机制。方法以腺样囊性癌ACC-2细胞株为研究对象。用转化生长因子β1(TGF-β1)以及ERK通路抑制剂U0126处理ACC-2细胞。MTT检测ACC-2细胞的增殖情况,Transwell实验检测细胞迁移、侵袭能力,Westernblot蛋白印迹检测ACC-2细胞中ERK1/2的活化及MMP-2的表达情况,实时荧光定量聚合酶链反应检测ACC-2细胞中MMP-2的mRNA的表达情况。结果 TGF-β1及U0126干预后,ACC-2细胞增殖能力无明显变化;TGF-β1刺激可增强ACC-2细胞迁移、侵袭能力,增加ACC-2细胞p-ERK1/2和MMP-2蛋白以及MMP-2mRNA的表达,而U0126阻断ERK磷酸化后,抑制了TGF-β1刺激的增强作用,ACC-2细胞的迁移、侵袭能力降低,MMP-2蛋白和mRNA的表达均下降。结论 TGF-β1/ERK1/2/MMP-2通路参与了人唾液腺ACC侵袭和迁移能力的调节。TGF-β1可通过上调ERK1/2,继而上调MMP-2,促进人唾液腺ACC细胞的侵袭和迁移能力,ERK1/2可能成为人唾液腺ACC侵袭防治的新靶点。

Objective To investigate the role of TGF-β1/ERK1/2/MMP-2 signaling pathways in the migration and invasion process of salivary adenoid cystic carcinoma cells.Methods ACC-2 cell line were used in this study.The proliferation of ACC-2 cell line was observed by MTT assay;cell migration and invasion ablity were examined by Transwell;the expression of MMP-2 in salivary adenoid cystic carcinoma cell lines was tested by Western blotting and Real-time PCR.The activity of ERK1/2 signaling pathway was measured by Western blotting.The effects of the ERK pathway specific inhibitors on TGF-β1-activated MMP-2 expression in ACC-2 cell lines were examined.Results TGF-β1 and U0126 could not enhance the proliferation of ACC-2 cells;TGF-β1 could enhance the migration and invasion abilities of them significantly.TGF-β1 enhanced expression of p-ERK1/2 and MMP-2 protein and MMP-2 mRNA in ACC-2 cells.When U0126 inhibited ERK1/2 pathway,the abilities of migration and invasion and production of MMP-2 stimulated by TGF-β1 were significantly inhibited in ACC-2 cells.Conclusions TGF-β1/ERK1/2/MMP-2 signaling pathways participate in the regulation of migration and invasion abilities in ACC.TGF-β1 can up-regulate ERK1/2,and then enhanced MMP-2 expression,promote the migration and invasion abilities of ACC.ERK1/2 may be turned to be a new target spot to prevent the invasion of ACC.