Chemokine CXCL12 activates dual CXCR4 and CXCR7-mediated signaling pathways in pancreatic cancer cells

Eileen L. Heinrich, Wendy Lee, Jianming Lu, Andrew M. Lowy, Joseph Kim

Research output: Contribution to journalArticlepeer-review

99 Scopus citations

Abstract

Background: Previously assumed to be a select ligand for chemokine receptor CXCR4, chemokine CXCL12 is now known to activate both CXCR4 and CXCR7. However, very little is known about the co-expression of these receptors in cancer cells.Methods: We used immunohistochemistry to determine the extent of co-expression in pancreatic cancer tissue samples and immunoblotting to verify expression in pancreatic cancer cell lines. In cell culture studies, siRNA was used to knock down expression of CXCR4, CXCR7, K-Ras and β-arrestin -2 prior to stimulating the cells with CXCL12. Activation of the mitogen-activated protein kinase pathway (MAPK) was assessed using both a Raf-pull down assay and western blotting. The involvement of the receptors in CXCL12-mediated increases in cell proliferation was examined via an ATP-based proliferation assay.Results: First, we discovered frequent CXCR4/CXCR7 co-expression in human pancreatic cancer tissues and cell lines. Next, we observed consistent increases in ERK1/2 phosphorylation after exposure to CXCL12 or CXCL11, a CXCR7 agonist, in pancreatic cancer cell lines co-expressing CXCR4/CXCR7. To better characterize the receptor-mediated pathway(s), we knocked down CXCR4 or CXCR7, exposed the cells to CXCL12 and examined subsequent effects on ERK1/2. We observed that CXCR7 mediates the CXCL12-driven increase in ERK1/2 phosphorylation. Knockdown of CXCR4 expression however, decreased levels of K-Ras activity. Conversely, KRAS knockdown greatly reduced CXCL12-mediated increases in ERK1/2 phosphorylation. We then evaluated the role of β-arrestin-2, a protein directly recruited by chemokine receptors. We observed that β-arrestin-2 knockdown also inhibited increases in ERK1/2 phosphorylation mediated by both CXCR4 and CXCR7. Finally, we investigated the mechanism for CXCL12-enhanced cell proliferation and found that either receptor can modulate cell proliferation.Conclusions: In summary, our data demonstrate that CXCR4 and CXCR7 are frequently co-expressed in human pancreatic cancer tissues and cell lines. We show that β-arrestin-2 and K-Ras dependent pathways coordinate the transduction of CXCL12 signals. Our results suggest that the development of therapies based on inhibiting CXCL12 signaling to halt the growth of pancreatic cancer should be focused at the ligand level in order to account for the contributions of both receptors to this signaling pathway.

Original languageEnglish
Article number68
JournalJournal of Translational Medicine
Volume10
Issue number1
DOIs
StatePublished - Apr 2 2012

Bibliographical note

Funding Information:
We appreciate the laboratory work of Xiaoming Shen, Ph.D. This work utilized City of Hope’s Pathology Core, which is supported in part by Cancer Center Support Grant NCI P30 CA33572. This study was also supported by NIH CA134637 (JK) and American Cancer Society RSG-11-070-01-TBE (JK). The contents are solely the responsibility of the authors and do not represent the official views of the NIH or ACS.

Keywords

  • CXCL12
  • CXCR4
  • CXCR7
  • MAPK
  • Pancreatic cancer

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology (all)

Fingerprint

Dive into the research topics of 'Chemokine CXCL12 activates dual CXCR4 and CXCR7-mediated signaling pathways in pancreatic cancer cells'. Together they form a unique fingerprint.

Cite this