Colorectal Surgery, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Braz J Med Biol Res. 2011 Jul;44(7):634-41. doi: 10.1590/s0100-879x2011007500070. Epub 2011 Jun 3.
The epithelial-mesenchymal transition (EMT) is involved in neoplastic metastasis, and the RON protein may be involved. In the present study, we determined the role and the mechanisms of action of RON in EMT in Madin-Darby canine kidney (MDCK) cells by Western blot and cell migration analysis. Activation of RON by macrophage stimulating protein (MSP) results in cell migration and initiates changes in the morphology of RON-cDNA-transfected MDCK cells. The absence of E-cadherin, the presence of vimentin and an increase in Snail were observed in RE7 cells, which were derived from MDCK cells transfected with wt-RON, compared with MDCK cells. Stimulation of RE7 cells with MSP resulted in increased migration (about 69% of the wounded areas were covered) as well as increased activation of extracellular signal-regulated kinase 1/2 (Erk1/2) and glycogen synthase kinase-3β (GSK-3β; the percent of the activation ratio was 143.6/599.8% and 512.4%, respectively), which could be inhibited with an individual chemical inhibitor PD98059 (50 μM) specific to MAPK/ERK kinase (the percent inhibition was 98.9 and 81.2%, respectively). Thus, the results indicated that RON protein could mediate EMT in MDCK cells via the Erk1/2 pathway. Furthermore, GSK-3β regulates the function of Snail in controlling EMT by this pathway.
上皮-间充质转化（EMT）参与肿瘤转移，RON 蛋白可能与之相关。在本研究中，我们通过 Western blot 和细胞迁移分析，确定了 RON 在 Madin-Darby 犬肾（MDCK）细胞 EMT 中的作用及其作用机制。巨噬细胞刺激蛋白（MSP）激活 RON 可导致细胞迁移，并引发 RON-cDNA 转染的 MDCK 细胞形态发生变化。与 MDCK 细胞相比，源自转染 wt-RON 的 MDCK 细胞的 RE7 细胞中观察到 E-钙黏蛋白缺失、波形蛋白存在以及 Snail 增加。MSP 刺激 RE7 细胞可导致迁移增加（约覆盖 69%的创面），同时细胞外信号调节激酶 1/2（Erk1/2）和糖原合酶激酶-3β（GSK-3β）的激活增加（激活率分别为 143.6/599.8%和 512.4%），这可被 MAPK/ERK 激酶的特异性化学抑制剂 PD98059（50 μM）抑制（抑制率分别为 98.9%和 81.2%）。因此，这些结果表明 RON 蛋白可通过 Erk1/2 通路介导 MDCK 细胞的 EMT。此外，GSK-3β 通过该通路调节 Snail 的功能以控制 EMT。
