Hypoxia-Induced Adaptations of miRNomes and Proteomes in Melanoma Cells and Their Secreted Extracellular Vesicles.

Geoffroy Walbrecq,Gunnar Dittmar,Odile Lecha,Miriam R Fougeras,Demetra Philippidou,François Bernardin,Christiane Margue,Iris Behrmann,Anthoula Gaigneaux,Milène Tetsi Nomigni,Stephanie Kreis

doi:10.3390/cancers12030692

Geoffroy Walbrecq, Gunnar Dittmar + Show 9 more

Open Access

https://doi.org/10.3390/cancers12030692

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Journal: Cancers	Publication Date: Mar 14, 2020
Citations: 32	License type: CC BY 4.0

Affiliation: University of Luxembourg, Luxembourg Institute of Health

Abstract

Reduced levels of intratumoural oxygen are associated with hypoxia-induced pro-oncogenic events such as invasion, metabolic reprogramming, epithelial–mesenchymal transition, metastasis and resistance to therapy, all favouring cancer progression. Small extracellular vesicles (EV) shuttle various cargos (proteins, miRNAs, DNA and others). Tumour-derived EVs can be taken up by neighbouring or distant cells in the tumour microenvironment, thus facilitating intercellular communication. The quantity of extracellular vesicle secretion and their composition can vary with changing microenvironmental conditions and disease states. Here, we investigated in melanoma cells the influence of hypoxia on the content and number of secreted EVs. Whole miRNome and proteome profiling revealed distinct expression patterns in normoxic or hypoxic growth conditions. Apart from the well-known miR-210, we identified miR-1290 as a novel hypoxia-associated microRNA, which was highly abundant in hypoxic EVs. On the other hand, miR-23a-5p and -23b-5p were consistently downregulated in hypoxic conditions, while the protein levels of the miR-23a/b-5p-predicted target IPO11 were concomitantly upregulated. Furthermore, hypoxic melanoma EVs exhibit a signature consisting of six proteins (AKR7A2, DDX39B, EIF3C, FARSA, PRMT5, VARS), which were significantly associated with a poor prognosis for melanoma patients, indicating that proteins and/or miRNAs secreted by cancer cells may be exploited as biomarkers.

Highlights

Melanoma is the most aggressive form of skin cancer with an annual increase of 0.6% estimated among adults over 50 years [1]
In order to get insights into the various molecular messengers operating in a hypoxic microenvironment in melanoma, the proteomes and miRNomes of whole cell lysates (WCLs) and
We determined the quality, purity and number of isolated extracellular vesicles (EV). nEVs and hEVs were isolated by ultracentrifugation from cell supernatants of four melanoma cell lines A375, 501Mel, MelJuso and IPC298

Summary

Introduction

Melanoma is the most aggressive form of skin cancer with an annual increase of 0.6% estimated among adults over 50 years [1]. Melanoma develops from melanocytes, which originate from neural crest-derived cells. Primary transformed melanocytes first form a melanoma in situ. The melanoma in situ will “create” an immunosuppressive environment, which allows the tumour cells to invade the lower dermis as well as blood or lymphatic vessels paving the way for metastatic disease [2]. With increasing size of the primary or metastatic tumour, cells in the centre experience a Cancers 2020, 12, 692; doi:10.3390/cancers12030692 www.mdpi.com/journal/cancers. Hypoxia has been demonstrated to be an aggravating factor in tumour development facilitating metastasis [4], e.g., by promoting lymph node metastasis in melanoma [5]. Hypoxia induces distinct changes in the tumour microenvironment (TME)

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