Gesualdi Luisa, Berardini Marika, Scicchitano Bianca Maria, Castaldo Clotilde, Bizzarri Mariano, Filippini Antonio, Riccioli Anna, Schiraldi Chiara, Ferranti Francesca, Liguoro Domenico, Mancini Rita, Ricci Giulia, Catizone Angela
Section of Histology and Medical Embryology, Department of Anatomy, Histology, Forensic-Medicine and Orthopedics, "Sapienza" University of Rome, 00161 Rome, Italy.
Section of Histology, Department of Life Sciences and Public Health, Università Cattolica del Sacro Cuore, 00168 Rome, Italy.
Biomedicines. 2023 Jul 4;11(7):1894. doi: 10.3390/biomedicines11071894.
c-MET/hepatocyte growth factor (HGF) system deregulation is a well-known feature of malignancy in several solid tumors, and for this reason this system and its pathway have been considered as potential targets for therapeutic purposes. In previous manuscripts we reported c-MET/HGF expression and the role in testicular germ cell tumors (TGCTs) derived cell lines. We demonstrated the key role of c-Src and phosphatidylinositol 3-kinase (PI3K)/AKT adaptors in the HGF-dependent malignant behavior of the embryonal carcinoma cell line NT2D1, finding that the inhibition of these onco-adaptor proteins abrogates HGF triggered responses such as proliferation, migration, and invasion. Expanding on these previous studies, herein we investigated the role of mitogen-activated protein kinase (MAPK)/extracellular signal regulated kinase (ERK) pathways in the HGF-dependent and HGF-independent NT2D1 cells biological responses. To inhibit MAPK/ERK pathways we chose a pharmacological approach, by using U0126 inhibitor, and we analyzed cell proliferation, collective migration, and chemotaxis. The administration of U0126 together with HGF reverts the HGF-dependent activation of cell proliferation but, surprisingly, does not exert the same effect on NT2D1 cell migration. In addition, we found that the use of U0126 alone significantly promotes the acquisition of NT2D1 «migrating phenotype», while collective migration of NT2D1 cells was stimulated. Notably, the inhibition of ERK activation in the absence of HGF stimulation resulted in the activation of the AKT-mediated pathway, and this let us speculate that the paradoxical effects obtained by using U0126, which are the increase of collective migration and the acquisition of partial epithelium-mesenchyme transition (pEMT), are the result of compensatory pathways activation. These data highlight how the specific response to pathway inhibitors, should be investigated in depth before setting up therapy.
c-MET/肝细胞生长因子（HGF）系统失调是几种实体瘤恶性肿瘤的一个众所周知的特征，因此该系统及其信号通路被认为是潜在的治疗靶点。在之前的论文中，我们报道了c-MET/HGF在睾丸生殖细胞肿瘤（TGCTs）衍生细胞系中的表达及其作用。我们证明了c-Src和磷脂酰肌醇3激酶（PI3K）/AKT衔接蛋白在胚胎癌细胞系NT2D1的HGF依赖性恶性行为中的关键作用，发现抑制这些癌衔接蛋白可消除HGF触发的增殖、迁移和侵袭等反应。在此基础上，我们进一步研究了丝裂原活化蛋白激酶（MAPK）/细胞外信号调节激酶（ERK）信号通路在HGF依赖性和非依赖性NT2D1细胞生物学反应中的作用。为了抑制MAPK/ERK信号通路，我们选择了一种药理学方法，即使用U0126抑制剂，并分析细胞增殖、集体迁移和趋化性。U0126与HGF联合使用可逆转HGF依赖性的细胞增殖激活，但令人惊讶的是，对NT2D1细胞迁移没有同样的作用。此外，我们发现单独使用U0126可显著促进NT2D1“迁移表型”的获得，同时刺激NT2D1细胞的集体迁移。值得注意的是，在没有HGF刺激的情况下抑制ERK激活会导致AKT介导的信号通路激活，这让我们推测，使用U0126获得的矛盾效应，即集体迁移增加和部分上皮-间质转化（pEMT）的获得，是补偿性信号通路激活的结果。这些数据凸显了在制定治疗方案之前，应深入研究对信号通路抑制剂的特异性反应。
