Increased migration of human mesenchymal stromal cells by autocrine motility factor (AMF) resulted in enhanced recruitment towards hepatocellular carcinoma.

Marcela Bolontrade,Guillermo Mazzolini,Flavia Piccioni,Osvaldo Podhajcer,Esteban Fiore,Juan Bayo,Mariana Malvicini,Manglio Rizzo,Jorge B Aquino,Mariana G Garcia,Estanislao Peixoto,Oscar Andriani,Laura Alaniz

doi:10.1371/journal.pone.0095171

Abstract

Background and AimsSeveral reports described the migration of human mesenchymal stromal cells (MSCs) towards tumor-released factors. Autocrine motility factor (AMF) is produced by several tumors including hepatocellular carcinoma (HCC). The aim of this study was to analyze AMF involvement on MSC migration towards human HCC.MethodsProduction of AMF by HCC tumors was evaluated by western analysis. The effects of AMF on MSCs from different sources (bone marrow, adipose tissue and perivascular cells from umbilical cord) were analyzed using in vitro migration assay; metalloproteinase 2 (MMP2) activity and expression of critical genes were studied by zymography and qRT-PCR, respectively. To assess AMF involvement on the in vivo MSC migration, noninvasive fluorescence imaging was performed. To test the effect of AMF-primed MSCs on tumor development, in vitro proliferation and spheroids growth and in vivo tumor volume were evaluated.ResultsAMF produced by HCC was found to induce migration of different MSCs in vitro and to enhance their MMP2 activity. Stimulation of MSCs with recombinant AMF (rAMF) also induced the in vitro adhesion to endothelial cells in coincidence with changes in the expression levels of MMP3, AMF receptor, caveolin-1, and -2 and GDI-2. Importantly, stimulation of MSCs with rAMF increased the in vivo migration of MSCs towards experimental HCC tumors. AMF-priming of MSCs did not induce a pro-tumorigenic effect on HCC cells neither in vivo nor in vitro.ConclusionAMF plays a role in MSC recruitment towards HCC. However, its ability to increase MSC migration to HCC for therapeutic purposes merits further evaluation.

Highlights

Hepatocellular carcinoma (HCC) is the sixth most common cancer worldwide and the third cause of cancer-related death [1]
A 55 KDa soluble Autocrine motility factor (AMF) form was detected in the conditioned medium of cell culture monolayers (CCM) and SC tumors (TCM) derived from HuH7 cell line and from HC-PT-5 hepatocellular carcinoma (HCC) primary cultures (Figure 1A)
Migration capacity of human mesenchymal stromal cells (MSCs) derived from bone marrow (BM-MSCs), perivascular umbilical cord (HUCPVCs) and adipose tissue (AT-MSCs) towards AMF was analyzed using a modified Boyden chamber assay

Summary

Introduction

Hepatocellular carcinoma (HCC) is the sixth most common cancer worldwide and the third cause of cancer-related death [1]. Curative therapies such as resection or liver transplantation improve patient survival [2]; these strategies can only be applied to a scarce minority of patients. Several reports described the use of MSCs as vehicles for therapeutic genes in cancer, exploiting the advantage of MSC homing into the tumor microenvironment [6]. Several reports described the migration of human mesenchymal stromal cells (MSCs) towards tumor-released factors. Autocrine motility factor (AMF) is produced by several tumors including hepatocellular carcinoma (HCC). The aim of this study was to analyze AMF involvement on MSC migration towards human HCC

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Conclusion