PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of brjcancerBJC HomepageBJC Advance online publicationBJC Current IssueSubmitting an article to BJCWeb feeds
 
Br J Cancer. 1999 March; 79(7-8): 1025–1031.
PMCID: PMC2362228

The chemokine RANTES is secreted by human melanoma cells and is associated with enhanced tumour formation in nude mice

Abstract

Modulation of tumour cell growth by tumour-infiltrating leucocytes is of high importance for the biological behaviour of malignant neoplasms. In melanoma, tumour-associated macrophages (TAM) and tumour-infiltrating lymphocytes (TIL) are of particular interest as inhibitors or enhancers of cell growth. Recruitment of leucocytes from the peripheral blood into the tumour site is mediated predominantly by chemotaxins, particularly by the group of chemokines.

The aim of this study was to identify peptides released by human melanoma cells with monocyte chemotactic properties. To assure the presence of biologically active mediators, biochemical purification and biological characterization of peptides was based on a detection system dependent on bioactive, monocyte chemotactic activity in vitro. Cell culture supernatants of melanoma cells were fractioned by heparin–sepharose followed by preparative reversed-phase HPLC steps to enrich monocyte chemotactic activity in one single band on a sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) gel. These purified fractions were shown to react with RANTES-specific antibodies in an enzyme-linked immunosorbent assay (ELISA) as well as in Western blot analysis. Amino acid sequencing of the N-terminal protein fragment confirmed 100% homology to the RANTES protein. Further analysis showed that four out of eight melanoma cell lines constitutively expressed and secreted the β-chemokine RANTES as detected by ELISA. The amount of RANTES protein secreted (up to 50 ng ml−1) was about 5–50 times higher than interleukin 8 (IL-8), determined in the same supernatant samples. Tumour necrosis factor α (TNF-α), not, however, IL-2, interferon-γ (IFN-γ), or α-melanocyte-stimulating hormone (α-MSH) was able to up-regulate RANTES and interleukin 8 secretion. Furthermore, higher levels of RANTES secretion in vitro were associated with increased tumour formation upon S.C. injection of six human melanoma cell lines in nude mice. Our data provide evidence that a subset of melanoma cells express mRNA and secrete RANTES protein which may be partly responsible for the recruitment of monocytes, T-cells and dendritic cells into the tumours. However, transplantation experiments in nude mice suggest that effects of RANTES may also benefit tumour progression. Further studies are needed to dissect the underlying mechanisms. © 1999 Cancer Research Campaign

Keywords: melanoma, metastasis, RANTES, chemokine, human

Full Text

The Full Text of this article is available as a PDF (103K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Artuc M, Nürnberg W, Czarnetzki BM, Schadendorf D. Characterization of gene regulatory elements for selective gene expression in human melanoma cells. Biochem Biophys Res Commun. 1995 Aug 15;213(2):699–705. [PubMed]
  • Barnhill RL. Pathology and prognostic factors. Curr Opin Oncol. 1993 Mar;5(2):364–376. [PubMed]
  • Benomar A, Ming WJ, Taraboletti G, Ghezzi P, Balotta C, Cianciolo GJ, Snyderman R, Doré JF, Mantovani A. Chemotactic factor and P15E-related chemotaxis inhibitor in human melanoma cell lines with different macrophage content and tumorigenicity in nude mice. J Immunol. 1987 Apr 1;138(7):2372–2379. [PubMed]
  • Bottazzi B, Polentarutti N, Acero R, Balsari A, Boraschi D, Ghezzi P, Salmona M, Mantovani A. Regulation of the macrophage content of neoplasms by chemoattractants. Science. 1983 Apr 8;220(4593):210–212. [PubMed]
  • Bröcker EB, Zwadlo G, Holzmann B, Macher E, Sorg C. Inflammatory cell infiltrates in human melanoma at different stages of tumor progression. Int J Cancer. 1988 Apr 15;41(4):562–567. [PubMed]
  • Cornil I, Theodorescu D, Man S, Herlyn M, Jambrosic J, Kerbel RS. Fibroblast cell interactions with human melanoma cells affect tumor cell growth as a function of tumor progression. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6028–6032. [PubMed]
  • Degli-Esposti MA, Davis-Smith T, Din WS, Smolak PJ, Goodwin RG, Smith CA. Activation of the lymphotoxin beta receptor by cross-linking induces chemokine production and growth arrest in A375 melanoma cells. J Immunol. 1997 Feb 15;158(4):1756–1762. [PubMed]
  • Ferrone S. Melanoma, immune surveillance, and immunotherapy. J Clin Invest. 1994 Apr;93(4):1351–1352. [PMC free article] [PubMed]
  • Gläser R, Rass K, Seiter S, Hauschild A, Christophers E, Tilgen W. Detection of circulating melanoma cells by specific amplification of tyrosinase complementary DNA is not a reliable tumor marker in melanoma patients: a clinical two-center study. J Clin Oncol. 1997 Aug;15(8):2818–2825. [PubMed]
  • Herlyn M, Thurin J, Balaban G, Bennicelli JL, Herlyn D, Elder DE, Bondi E, Guerry D, Nowell P, Clark WH, et al. Characteristics of cultured human melanocytes isolated from different stages of tumor progression. Cancer Res. 1985 Nov;45(11 Pt 2):5670–5676. [PubMed]
  • Mattei S, Colombo MP, Melani C, Silvani A, Parmiani G, Herlyn M. Expression of cytokine/growth factors and their receptors in human melanoma and melanocytes. Int J Cancer. 1994 Mar 15;56(6):853–857. [PubMed]
  • Rigel DS, Friedman RJ, Kopf AW. The incidence of malignant melanoma in the United States: issues as we approach the 21st century. J Am Acad Dermatol. 1996 May;34(5 Pt 1):839–847. [PubMed]
  • Rivers JK. Melanoma. Lancet. 1996 Mar 23;347(9004):803–806. [PubMed]
  • Schadendorf D, Möller A, Algermissen B, Worm M, Sticherling M, Czarnetzki BM. IL-8 produced by human malignant melanoma cells in vitro is an essential autocrine growth factor. J Immunol. 1993 Sep 1;151(5):2667–2675. [PubMed]
  • Schall TJ, Bacon KB. Chemokines, leukocyte trafficking, and inflammation. Curr Opin Immunol. 1994 Dec;6(6):865–873. [PubMed]
  • Singh RK, Varney ML. Regulation of interleukin 8 expression in human malignant melanoma cells. Cancer Res. 1998 Apr 1;58(7):1532–1537. [PubMed]
  • Sozzani S, Sallusto F, Luini W, Zhou D, Piemonti L, Allavena P, Van Damme J, Valitutti S, Lanzavecchia A, Mantovani A. Migration of dendritic cells in response to formyl peptides, C5a, and a distinct set of chemokines. J Immunol. 1995 Oct 1;155(7):3292–3295. [PubMed]
  • Sticherling M, Schröder JM, Christophers E. Production and characterization of monoclonal antibodies against the novel neutrophil activating peptide NAP/IL-8. J Immunol. 1989 Sep 1;143(5):1628–1634. [PubMed]
  • Sticherling M, Küpper M, Koltrowitz F, Bornscheuer E, Kulke R, Klinger M, Wilhelm D, Kameyoshi Y, Christophers E, Schröder JM. Detection of the chemokine RANTES in cytokine-stimulated human dermal fibroblasts. J Invest Dermatol. 1995 Oct;105(4):585–591. [PubMed]
  • van Ravenswaay Claasen HH, Kluin PM, Fleuren GJ. Tumor infiltrating cells in human cancer. On the possible role of CD16+ macrophages in antitumor cytotoxicity. Lab Invest. 1992 Aug;67(2):166–174. [PubMed]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK