Endothelin A receptor drives invadopodia function and cell motility through the β-arrestin/PDZ-RhoGEF pathway in ovarian carcinoma

• Published in: Oncogene Vol. 35; no. 26; pp. 3432 - 3442
• Main Authors: Semprucci, E, Tocci, P, Cianfrocca, R, Sestito, R, Caprara, V, Veglione, M, Castro, V Di, Spadaro, F, Ferrandina, G, Bagnato, A, Rosanò, L
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 30.06.2016; Nature Publishing Group
• Subjects: 38; 42/35; 45; 45/29; 59; 631/67; 631/67/1517/1709; 64; 64/60; 96; Actin; Actin Depolymerizing Factors - metabolism; Actins - metabolism; Adaptor Proteins, Vesicular Transport - metabolism; Animals; Apoptosis; Arrestin; beta-Arrestins - genetics; beta-Arrestins - metabolism; Cell Biology; Cell Line, Tumor; Cell Movement - genetics; Cell Movement - physiology; Cell receptors; Cellular signal transduction; Cofilin; Cortactin - metabolism; Development and progression; Endothelin 1; Endothelin ETA receptors; Female; Genetic aspects; Health aspects; Human Genetics; Humans; Immunoblotting; Internal Medicine; Invasiveness; Kinases; Lim Kinases - metabolism; Matrix metalloproteinase; Matrix Metalloproteinase 14 - metabolism; Medicine; Medicine & Public Health; Metalloproteinase; Metastases; Metastasis; Mice, Nude; Motility; Oncology; original-article; Ovarian cancer; Ovarian carcinoma; Ovarian Neoplasms - genetics; Ovarian Neoplasms - metabolism; Ovarian Neoplasms - pathology; Phosphorylation; Podosomes - genetics; Podosomes - metabolism; Podosomes - physiology; Postsynaptic density; Properties; Receptor, Endothelin A - genetics; Receptor, Endothelin A - metabolism; Reverse Transcriptase Polymerase Chain Reaction; rho GTP-Binding Proteins - metabolism; Rho Guanine Nucleotide Exchange Factors - genetics; Rho Guanine Nucleotide Exchange Factors - metabolism; rho-Associated Kinases - metabolism; RhoA protein; rhoC GTP-Binding Protein; RNA Interference; Signal transduction; Signal Transduction - genetics; Transplantation, Heterologous; Tumors
• ISSN: 0950-9232; 1476-5594
• DOI: 10.1038/onc.2015.403

The endothelin-1 (ET-1)/ET A receptor (ET A R) signalling pathway is a well-established driver of epithelial ovarian cancer (EOC) progression. One key process promoted by ET-1 is tumor cell invasion, which requires the scaffolding functions of β-arrestin-1 (β-arr1) downstream of the receptor; however, the potential role of ET-1 in inducing invadopodia, which are crucial for cellular invasion and tumor metastasis, is completely unknown. We describe here that ET-1/ET A R, through β-arr1, activates RhoA and RhoC GTPase and downstream ROCK (Rho-associated coiled coil-forming kinase) kinase activity, promoting actin-based dynamic remodelling and enhanced cell invasion. This is accomplished by the direct interaction of β-arr1 with PDZ-RhoGEF ( p ostsynaptic density protein 95/ d isc- l arge/ z onula occludens-RhoGEF). Interestingly, ET A R-mediated invasive properties are related to the regulation of invadopodia, as evaluated by colocalization of actin with cortactin, as well as with TKS5 and MT1-MMP (membrane type 1-matrix metalloproteinase) with areas of matrix degradation, and activation of cofilin pathway, which is crucial for regulating invadopodia activity. Depletion of PDZ-RhoGEF, or β-arr1, or RhoC, as well as the treatment with the dual ET-1 receptor antagonist macitentan, significantly impairs invadopodia function, MMP activity and invasion, demonstrating that β-arr1/PDZ-RhoGEF interaction mediates ET A R-driven ROCK-LIMK-cofilin pathway through the control of RhoC activity. In vivo , macitentan is able to inhibit metastatic dissemination and cofilin phosphorylation. Collectively, our data unveil a noncanonical activation of the RhoC/ROCK pathway through the β-arr1/PDZ-RhoGEF complex as a regulator of ET A R-induced motility and metastasis, establishing ET-1 axis as a novel regulator of invadopodia protrusions through the RhoC/ROCK/LIMK/cofilin pathway during the initial steps of EOC invasion.