Mambalgin-2 Inhibits Growth, Migration, and Invasion of Metastatic Melanoma Cells by Targeting the Channels Containing an ASIC1a Subunit Whose Up-Regulation Correlates with Poor Survival Prognosis.

Maxim L Bychkov,Ekaterina N Lyukmanova,Artem V Kirichenko,Mikhail A Shulepko,Mikhail P Kirpichnikov,Irina N Mikhaylova

doi:10.3390/biomedicines9101324

Maxim L Bychkov, Ekaterina N Lyukmanova + Show 4 more

Open Access

https://doi.org/10.3390/biomedicines9101324

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Abstract

Melanoma is an aggressive cancer characterized by the acidification of the extracellular environment. Here, we showed for the first time that extracellular media acidification increases proliferation, migration, and invasion of patient-derived metastatic melanoma cells and up-regulates cell-surface expression of acid-sensitive channels containing the ASIC1a, α-ENaC, and γ-ENaC subunits. No influence of media acidification on these processes was found in normal keratinocytes. To control metastatic melanoma progression associated with the ASIC1a up-regulation, we proposed the ASIC1a inhibitor, -mambalgin-2 from Dendpoaspis polylepis venom. Recombinant analog of mambalgin-2 cancelled acidification-induced proliferation, migration, and invasion of metastatic melanoma cells, promoted apoptosis, and down-regulated cell-surface expression of prooncogenic factors CD44 and Frizzled 4 and phosphorylation of transcription factor SNAI. Confocal microscopy and affinity purification revealed that mambalgin-2 interacts with heterotrimeric ASIC1a/α-ENaC/γ-ENaC channels on the surface of metastatic melanoma cells. Using the mutant variant of mambalgin-2 with reduced activity toward ASIC1a, we confirmed that the principal molecular target of mambalgin-2 in melanoma cells is the ASIC1a subunit. Bioinformatic analysis confirmed up-regulation of the ASIC1 expression as a marker of poor survival prognosis for patients with metastatic melanoma. Thus, targeting ASIC1a by drugs such as mambalgin-2 could be a promising strategy for metastatic melanoma treatment.

Highlights

Melanoma is a very heterogeneous form of cancer, characterized by severe proliferation, invasion, and drug resistance [1]
We showed previously that apoptosis induction underlies the mambalgin-2 antiproliferative action in GBM cells [23], and propose here that the growth inhibition due to the apoptosis induction is common for the mambalgin-2 mediated ASIC1 inhibition in cancer cells
We showed that acidification of extracellular media drives proliferation, migration, and invasion of patient-derived metastatic melanoma cells, but not of normal keratinocytes

Summary

Introduction

Melanoma is a very heterogeneous form of cancer, characterized by severe proliferation, invasion, and drug resistance [1] All these processes are accompanied by induction of hypoxia and elevated extracellular acidification [2,3]. After adaptation to the acidic conditions, melanoma cells exhibit the upregulation of signaling pathways important for tissue remodeling, cell cycle progression, proliferation, and motility, even if the extracellular pH returns to physiological values [6,7]. Another facet of the melanoma extracellular media acidification is induction of the epithelial-mesenchymal transition (EMT). Acidification of the extracellular media may drive growth, migration, and invasion of melanoma cells; and targeting of the molecules, responsible for melanoma cell adaptation to the acidic conditions can be a perspective strategy for melanoma therapy

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