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ERK Signaling Pathway Is Constitutively Active in NT2D1 Non-Seminoma Cells and Its Inhibition Impairs Basal and
Luisa Gesualdi1, Marika Berardini1, Bianca Maria Scicchitano2,3
1Section of Histology and Medical Embryology, Department of Anatomy, Histology, Forensic-Medicine and Orthopedics, "Sapienza" University of Rome, 00161 Rome, Italy.
Abstract:
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.
Insights
Deregulation of the c-MET/hepatocyte growth factor (HGF) pathway is implicated in cancer. This study reveals that inhibiting MAPK/ERK pathways in testicular germ cell tumors paradoxically enhances migration, suggesting complex compensatory mechanisms that warrant careful therapeutic consideration.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Deregulation of the c-MET/hepatocyte growth factor (HGF) system is common in various solid tumors, making it a therapeutic target.
- Previous research established the role of c-MET/HGF and associated signaling pathways (c-Src, PI3K/AKT) in testicular germ cell tumors (TGCTs), specifically the NT2D1 cell line.
Purpose of the Study:
- To investigate the involvement of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathways in both HGF-dependent and HGF-independent biological responses of NT2D1 cells.
- To analyze the effects of inhibiting MAPK/ERK pathways on cell proliferation, collective migration, and chemotaxis in TGCT models.
Main Methods:
- Pharmacological inhibition of MAPK/ERK pathways using the U0126 inhibitor.
- Assessment of cell proliferation, collective migration, and chemotaxis in NT2D1 cells under various conditions (HGF stimulation, U0126 treatment, or both).
- Analysis of downstream signaling pathways, including AKT activation, to understand compensatory mechanisms.
Main Results:
- U0126 treatment, combined with HGF, reversed HGF-dependent proliferation but did not affect cell migration.
- Unexpectedly, U0126 alone promoted NT2D1 cell migration and collective migration.
- Inhibition of ERK activation without HGF stimulation led to AKT pathway activation, suggesting compensatory mechanisms driving migration and partial epithelial-mesenchymal transition (pEMT).
Conclusions:
- The MAPK/ERK pathway plays a complex role in TGCT cell behavior, with inhibition leading to paradoxical pro-migratory effects.
- Compensatory pathway activation (e.g., AKT) following ERK inhibition can drive increased migration and pEMT.
- These findings underscore the necessity for in-depth investigation of cellular responses to pathway inhibitors before clinical application in cancer therapy.
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