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Transcriptional profiling identifies genes induced by hepatocyte-derived extracellular matrix in metastatic human colorectal cancer cell lines

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Abstract

The milieu of the liver, and in particular hepatocyte-derived extracellular matrix (hECM), is a critical factor regulating development of liver metastases of colorectal cancer (CRC) cells. The present study has investigated genes altered by hECM in CRC cells and particularly by heparan sulfate chains of hepatocyte proteoglycans. Gene profiling analysis shows that after 2 days on hECM, 226 genes are up-regulated more than 2-fold in strongly metastatic SM cells, including genes involved in growth arrest and apoptosis, signal transduction, cell migration, proliferation, communication and angiogenesis, with activation of the erbB signaling network and p53 effectors. Genes down-regulated by hECM include genes involved in lipogenesis and the S phase of the cell cycle. Further studies exploring the kinetics of gene expression after 4 and 7 days culture on hECM show induction of EGF family members and of stem cell markers. In particular, hECM, but not collagen, increases mRNA expression of HB-EGF and colon stem cell marker leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5). Expression of these genes is not induced by hECM depleted of the heparan sulfate chains of proteoglycans. Lastly, a specific cell population positive for cancer stem cell (CSC) markers LGR5, epCAM and CD133, but negative for CD44, appears after 7 days culture on hECM, a population which is reduced by 50 % in cells grown on heparan sulfated-depleted hECM. Collectively, the data suggest that hECM induces growth factors and receptors regulating proliferation of metastatic CRC in the liver and offers a growth advantage for specific populations expressing CSC markers.

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Abbreviations

AREG:

Amphiregulin

CCNG2:

Cyclin G2

CDH1:

E-cadherin

CRC:

Colorectal cancer

CSC:

Cancer stem cell

DSP:

Desmoplakin

EGFR:

EGF receptor

EMT:

Epithelial- to mesenchymal transition

epCAM:

Epithelial cell adhesion molecule

EREG:

Epiregulin

FCS:

Fetal calf serum

HB-EGF:

Heparin-binding EGF-like growth factor

HDM:

Hormonally defined medium

hECM:

Hepatocyte-derived extracellular matrix

LGR5:

Leucine-rich repeat-containing G-protein-coupled receptor 5

LPN2:

Lipocalin 2

MET:

Mesenchymal- to epithelial transition

Mod hECM:

Heparan sulfate-depleted hECM

MMP-10:

Metalopeptidase 10, stromelysin 2

SM:

KM12SM (strongly metastatic CRC cell line)

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Acknowledgments

This project was supported by an internal Grant from Tel Aviv Sourasky Medical Center to Dr. R. Kariv (courtesy of I. Ben-Dov) and by an internal grant from Tel Aviv University to Dr. R Kariv and Dr I. Zvibel.

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The authors declare that they have no conflicts of interest.

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Correspondence to Revital Kariv.

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Isabel Zvibel and Adam Wagner contributed equally to this work.

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Zvibel, I., Wagner, A., Pasmanik-Chor, M. et al. Transcriptional profiling identifies genes induced by hepatocyte-derived extracellular matrix in metastatic human colorectal cancer cell lines. Clin Exp Metastasis 30, 189–200 (2013). https://doi.org/10.1007/s10585-012-9527-8

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