Clinical & Experimental Metastasis

, Volume 24, Issue 2, pp 121–130 | Cite as

Chemokine expression is associated with the accumulation of tumour associated macrophages (TAMs) and progression in human colorectal cancer

  • Charles Bailey
  • Rupert Negus
  • Alistair Morris
  • Paul Ziprin
  • Robert Goldin
  • Paola Allavena
  • David Peck
  • Ara Darzi
Research Paper

Abstract

Chemokines promote tumour progression by enhancing proliferation and modifying the immune response. The purpose of this study was to test the hypothesis that CCL2 monocyte chemotactic protein-1 (MCP-1) contributes to the progression of colorectal cancer by influencing the number and distribution of tumour associated macrophages (TAMs). Chemokine expression was assessed in human colorectal adenocarcinomas by ribonuclease protection assay (RPA). Colonic adenocarcinoma cell lines were used to assess chemokine production by enzyme linked immunosorbant assay (ELISA), and Boyden microchemotaxis assays were performed to determine cell line supernatant monocyte chemotactic activity. CCL2 production was assessed in paraffin embedded tumour samples by immunohistochemistry. Finally, the number of macrophages and their distribution was determined in the same colorectal adenocarcinomas and compared with CCL2 expression and tumour stage. Results showed that CCL2 produced by cell lines induced monocyte chemoattraction, the expression of this chemokine in solid cancers increased with tumour stage (P < 0.05) and immunohistochemistry localized production to tumour cells. Analysis of the macrophage infiltrate showed that the accumulation was significantly greater in tumours than controls (P < 0.005) and within tumours it was greatest in necrotic regions (median 44,600 per mm3). Macrophage accumulation increased with tumour stage and correlated with CCL2 expression (rs = 0.8). CXCL8 interleukin 8 (IL-8), a potent angiogenic factor and growth factor, was expressed in all tumours and cell lines. It is concluded that CCL2 induces the accumulation of tumour promoting TAMs in human colorectal cancer and represents a therapeutic target to modify the macrophage response and direct immune mediated therapy.

Keywords

Tumour associated macrophage (TAM) Leukocyte Cytokine Chemokine CCL2 (MCP-1) CXCL8 (IL-8) Colon cancer Rectal cancer 

Abbreviations

ATP

Adenosine triphosphate

DAB

Diaminobenzidine

ELISA

Enzyme linked immunosorbant assay

FBS

Fetal bovine serum

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase

IL-8

Interleukin 8

IP-10

Interferon inducible protein 10

Ltn

Lymphotactin

mAb

Monoclonal antibody

MCP-1

Monocyte chemotactic protein-1

MIP-1α

Macrophage inflammatory protein 1 alpha

MIP-1β

Macrophage inflammatory protein 1 beta

MMP-9

Matrix metalloproteinase-9

PBS

Phosphate buffered saline

RANTES

Regulated on activation normal T-cell expressed and secreted

RPA

Ribonuclease protection assay

SE

Standard error

TAM

Tumour associated macrophage

TBE

Tris-borate buffer

TNFα

Tumour necrosis factor α

UTP

Uridine-5′-triphosphate

VEGF

Vascular endothelial growth factor

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Copyright information

© Springer Science + Business Media B.V. 2007

Authors and Affiliations

  • Charles Bailey
    • 1
    • 2
  • Rupert Negus
    • 3
  • Alistair Morris
    • 1
  • Paul Ziprin
    • 1
  • Robert Goldin
    • 4
  • Paola Allavena
    • 5
  • David Peck
    • 1
  • Ara Darzi
    • 1
  1. 1.Department of Surgical Oncology and TechnologySt Mary’s Hospital, Imperial College of Science, Technology, and MedicineLondonUK
  2. 2.Department of Colorectal SurgeryLevel 2, John Radcliffe HospitalOxfordUK
  3. 3.Department of GastroenterologySt Mary’s HospitalLondonUK
  4. 4.Department of PathologySt Mary’s Hospital, Imperial College of Science, Technology, and MedicineLondonUK
  5. 5.Istituto Clinico HumanitasRozzano, MilanItaly

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