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. 1994 Jul;39(4):231–238. doi: 10.1007/BF01525986

Expression of theJE/MCP-1 gene suppresses metastatic potential in murine colon carcinoma cells

Suyun Huang 1, Rakesh K Singh 1, Keping Xie 1, Mordechai Gutman 1, Karen K Berry 1, Corazon D Bucana 1, Isaiah J Fidler 1, Menashe Bar-Eli 1,✉
PMCID: PMC11038689  PMID: 7954525

Abstract

The purpose of this study was to determine whether the expression of theJE/MCP-1 gene encoding for the monocyte chemottractant protein, MCP-1 (also known as monocyte chemotactic and activating factor MCAF, TDCF, and SMC-CF) can influence the metastatic properties of tumor cells. The highly metastatic murine colon carcinoma CT-26 cells, syngeneic to BALB/c mice that do not produce endogenous JE/MCP-1 protein, were transfected with a BCMGS-Neo expression vector (control) or a vector containing full-lengthJE cDNA. CT-26 parental cells, CT-26 Neo, and CT-26 JE/MCP-1-positive cells were injected into syngeneic or nude mice. The CT-26 JE/MCP-1-positive cells produced significantly fewer lung metastases. The decrease in incidence of metastasis was not due to the inability of the transfected cells to arrest in the lung vasculature or to differences in cell cycle time. CT-26 cells producing JE/MCP-1 were highly susceptible to lysis by syngeneic macrophages treated with subthreshold concentrations of lipopolysaccharide. In addition, culture supernatants of JE/MCP-1-expressing cells plus lipopolysaccharide synergistically activated tumoricidal properties in syngeneic macrophages. This activity was blocked by anti-JE/MCP-1 antibodies, indicating the involvement of the JE/MCP-1 molecule in this process. Moreover, purified JE/MCP-1 added to lipopolysaccharide-containing medium resulted in significant activation of macrophages against parental CT-26 cells. These data suggest that, in addition to its chemotactic properties, JE/MCP-1 can synergize with bacterial endotoxins to activate macrophages to become tumoricidal and, hence, could suppress metastasis.

Key words: Metastasis, Cytokines, Colon carcinoma

References

  • 1.Bar-Eli M, Gallily R. The effect of macrophage hydrolytic enzyme levels on the uptake and degradation of antigen and immune complexes. J Reticuloendothel Soc. 1975;18:317. [PubMed] [Google Scholar]
  • 2.Benomar A, Wang JM, Taraboletti G, Ghezzi P, Balotta C, Cianciolo GJ, Snyderman R, Dore JF, Mantovani A. Chemotactic factors and P15E-related chemotaxis inhibitor in human melanoma cell lines with different macrophage content and tumorigenicity in nude mice. J Immunol. 1987;136:2372. [PubMed] [Google Scholar]
  • 3.Bottazzi B, Polentarutti N, Balsari A, Boraschi D, Ghezzi P, Salmona M, Mantovani A. Chemotactic activity for mononuclear phagocytes of culture supernatants from murine and human tumor cells: evidence for a role in the regulation of macrophage content of neoplastic tissues. Int J Cancer. 1983;31:55. doi: 10.1002/ijc.2910310110. [DOI] [PubMed] [Google Scholar]
  • 4.Bottazzi B, Polentarutti N, Acero R, Balsari A, Boraschi D, Ghezzi P, Salmona M, Mantovani A. Regulation of macrophage content of neoplasms by chemoattractants. Science. 1983;220:210. doi: 10.1126/science.6828888. [DOI] [PubMed] [Google Scholar]
  • 5.Bottazzi B, Ghezzi P, Taraboletti G, Salmona M, Colombo N, Bonazzi C, Mangioni C, Mantovani A. Tumor-derived chemotactic factor(s) from human ovarian carcinoma: evidence for a role in the regulation of the macrophage content of neoplastic tissues. Int J Cancer. 1985;36:167. doi: 10.1002/ijc.2910360207. [DOI] [PubMed] [Google Scholar]
  • 6.Bottazzi B, Walter S, Govoni D, Collota F, Mantovani A. Monocyte chemotactic cytokine gene transfer modulates macrophage infiltration, growth and susceptibility to IL-2 therapy of a murine melanoma. J Immunol. 1992;148:1280. [PubMed] [Google Scholar]
  • 7.Bottazzi B, Colotta F, Sica A, Nobili N, Mantovani A. A chemoattractant expressed in human sarcoma cell (tumor-derived chemotactic factor, TCDF) is identical to monocyte chemoattractant protein-1/monocyte chemotactic and activating factor (MCP-1/MCAF) Int J Cancer. 1990;45:795. doi: 10.1002/ijc.2910450436. [DOI] [PubMed] [Google Scholar]
  • 8.Cochran BH, Reffel AC, Stiles CD. Molecular cloning of gene sequences regulated by platelet-derived growth factor. Cell. 1983;33:939. doi: 10.1016/0092-8674(83)90037-5. [DOI] [PubMed] [Google Scholar]
  • 9.Corbett TH, Griswold DP, Jr, Roberts BJ. Tumor induction relationships in development of transplantable cancers of the colon in mice for chemotherapy assays with a note on carcinogen structure. Cancer Res. 1975;23:2434. [PubMed] [Google Scholar]
  • 10.Drysdale BE, Agarwal S, Shin HS. Macrophage-mediated tumoricidal activity: mechanisms of activation and cytotoxicity. Prog Allergy. 1988;40:111. [PubMed] [Google Scholar]
  • 11.Esumi N, Fan D, Fidler IJ. Inhibition of murine melanoma experimental metastasis by recombinant desulfatohirudin, a highly specific thrombin inhibitor. Cancer Res. 1991;51:4549. [PubMed] [Google Scholar]
  • 12.Fidler IJ. Metastasis: quantitative analysis of distribution and fate of tumor emboli labeled with125I-5-iodo-2′-deoxyuridine. J Natl Cancer Inst. 1970;45:773. [PubMed] [Google Scholar]
  • 13.Fidler IJ. Macrophages and metastasis—a biological approach to cancer therapy: presidential address. Cancer Res. 1985;45:4714. [PubMed] [Google Scholar]
  • 14.Fidler IJ. Rationale and methods for the use of nude mice to study the biology and therapy of human cancer metastasis. Cancer Metastasis Rev. 1986;5:29. doi: 10.1007/BF00049529. [DOI] [PubMed] [Google Scholar]
  • 15.Fidler IJ. Critical factors in the biology of human cancer metastasis: twenty-eighth G.H.A. Clowes Memorial Award Lecture. Cancer Res. 1990;50:6130. [PubMed] [Google Scholar]
  • 16.Fidler IJ, Gersten DM, Kripke ML. The influence of immune status on the metastasis of three murine fibrosarcomas of different immunogenicities. Cancer Res. 1979;39:3816. [PubMed] [Google Scholar]
  • 17.Furutani Y, Nomura H, Notake M, Oyamada Y, Fukui T, Yamada M, Larsen CGG, Oppenheim JJ, Matsushima K. Cloning and sequencing of the cDNA for human monocyte chemotactic and activating factor (MCAF) Biochem Biophys Res Commun. 1989;159:249. doi: 10.1016/0006-291x(89)92430-3. [DOI] [PubMed] [Google Scholar]
  • 18.Gallily R, Bar-Eli M. Studies on the relationships between the immunogenicity and catabolism of antigens and their binding to the surface of macrophages. Eur J Immunol. 1976;6:789. doi: 10.1002/eji.1830061107. [DOI] [PubMed] [Google Scholar]
  • 19.Graves D, Jiang TYL, Williamson A, Valente AJ. Identification of monocyte chemotactic activity produced by malignant cells. Science. 1989;245:1490. doi: 10.1126/science.2781291. [DOI] [PubMed] [Google Scholar]
  • 20.Huang S, Paulauskis JD, Godleski JJ, Kobzik L. Expression of macrophages inflammatory protein-2 and KC mRNA in pulmonary inflammation. Am J Pathol. 1992;141:981. [PMC free article] [PubMed] [Google Scholar]
  • 21.Luca M, Hunt B, Bucana CD, Johnson JP, Fidler IJ, Bar-Eli M. Direct correlation between MUC18 expression and metastatic potential of human melanoma cells. Melanoma Res. 1993;3:35. doi: 10.1097/00008390-199304000-00006. [DOI] [PubMed] [Google Scholar]
  • 22.Mantovani A, Bottazzi B, Colotta F, Sozzani S, Ruco L. The origin and function of tumor associated macrophages. Immunol Today. 1992;13:265. doi: 10.1016/0167-5699(92)90008-U. [DOI] [PubMed] [Google Scholar]
  • 23.Matsushima K, Larsen CG, Dubois CG, Oppenheim JJ. Purification and characterization of a novel monocyte chemotactic protein and activating factor produced by a human myelo-monocytic cell line. J Exp Med. 1989;169:1485. doi: 10.1084/jem.169.4.1485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Oppenheim JJ, Zachariae COC, Mukaida N, Matsushima K. Properties of the novel proinflammatory supergene “intercrine” cytokine family. Annu Rev Immunol. 1991;9:617. doi: 10.1146/annurev.iy.09.040191.003153. [DOI] [PubMed] [Google Scholar]
  • 25.Raz A, Fogler WE, Fidler IJ. The effect of experimental condition on the expression of in vitro tumor cytotoxicity mediated by murine macrophages. Cancer Immunol Immunother. 1979;7:157. [Google Scholar]
  • 26.Rollins BJ, Sunday ME. Suppression of tumor formation in vivo by expression of the JE genes in malignant cells. Mol Cell Biol. 1991;11:3125. doi: 10.1128/mcb.11.6.3125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.Rollins BJ, Morrison ED, Stiles CD. Cloning and expression of JE, a gene inducible by platet-derived growth factor and whose product has cytokine-like properties. Proc Natl Acad Sci USA. 1988;85:3738. doi: 10.1073/pnas.85.11.3738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28.Saiki I, Fidler IJ. Synergistic activation of recombinant mouse gamma interferon and muramyl dipeptide of tumoricidal properties in mouse macrophages. J Immunol. 1979;135:604. [PubMed] [Google Scholar]
  • 29.Schackert H, Fidler IJ. Development of an animal model to study the biology of recurrent colorectal cancer originating from mesenteric lymph system metastasis. Int J Cancer. 1989;44:177. doi: 10.1002/ijc.2910440131. [DOI] [PubMed] [Google Scholar]
  • 30.Singh RK, Berry K, Matsushima N, Mukaida N, Fidler IJ. Synergism between human monocyte chemotactic and activating factor (MCAF) and bacterial products for activation of tumoricidal properties in murine macrophages. J Immunol. 1993;151:2786. [PubMed] [Google Scholar]
  • 31.Sone S, Fidler IJ. In situ activation of tumoricidal properties in rat alveolar macrophages and rejection of experimental lung metastases by intravenous injections ofNocardia rubra cell wall skeleton. Cancer Immunol Immunother. 1982;9:227. [Google Scholar]
  • 32.Van Damme J, Decock B, Lenaerts JP, Conings R, Bertini R, Mantovani A, Billiau A. Identification by sequence analysis of chemotactic factors for monocytes produced by normal transformed cells stimulated with virus, double stranded RNA or IL-1. Eur J Immunol. 1989;19:2367. doi: 10.1002/eji.1830191228. [DOI] [PubMed] [Google Scholar]
  • 33.Van Damme J, Decok B, Bertini R, Conings R, Leanerts JP, Put W, Opdenakker G, Mantovani A. Production and identification of natural monocyte chemotactic protein (MCP) from virally infected fibroblasts. Relationship with the JE gene product. Eur J Biochem. 1991;199:223. doi: 10.1111/j.1432-1033.1991.tb16113.x. [DOI] [PubMed] [Google Scholar]
  • 34.Walter S, Bottazi B, Govoni D, Colotta F, Mantovani A. Macrophage infiltration and growth of sarcoma clones expressing different amounts of monocyte chemotactic protein/JE. Int J Cancer. 1991;49:431. doi: 10.1002/ijc.2910490321. [DOI] [PubMed] [Google Scholar]
  • 35.Wang JM, Cianciolo GJ, Snyderman R, Mantovani A. Coexistence of a chemotactic factor and a retroviral P15E-related chemotaxis inhibitor in human tumor cell culture supernatants. J Immunol. 1986;137:2726. [PubMed] [Google Scholar]
  • 36.Widner U, Manogua KR, Cerami A, Sherry B. Genomic cloning and promoter analysis of macrophage inflammatory protein (MIP-2, MIP-1, and MIP-1-beta, members of the chemokine superfamily of proinflammatory cytokines. J Immunol. 1993;150:11. [PubMed] [Google Scholar]
  • 37.Xia YY, Feng LL, Yoshimura T, Wilson CB. LPS-induced MCP-1, IL-1-beta, and TNF-alpha messenger RNA expression in isolated erythrocyte-perfused rat kidney. Am J Physiol. 1993;264:5. doi: 10.1152/ajprenal.1993.264.5.F774. [DOI] [PubMed] [Google Scholar]
  • 38.Yoshimura T, Robinson EA, Tanaka S, Appella E, Kuratsu J-I, Leonard EJ. Purification and amino acid analysis of two glioma-derived monocyte chemoattractants. J Exp Med. 1989;169:1449. doi: 10.1084/jem.169.4.1449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 39.Yoshimura T, Yuhki N, Moore SK, Appella E, Lerman M, Leonard EJ. Human monocyte chemoattractant protein-1 (MCP-1): Full-length cDNA cloning, expression in mitogen-stimulated bloodmononuclear leukocytes, and sequence similarity to mouse competence gene JE. FEBS Lett. 1989;244:487. doi: 10.1016/0014-5793(89)80590-3. [DOI] [PubMed] [Google Scholar]
  • 40.Zachariae COC, Anderson AO, Thompson HL, Appella E, Mantovani A, Oppenheim JJ, Matsushima K. Properties of monocyte chemotactic and activating factor (MCAF) purified from a human fibrosarcoma cell line. J Exp Med. 1990;171:2177. doi: 10.1084/jem.171.6.2177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 41.Zhang RD, Price JE, Schackert G, Itoh K, Fidler IJ. Malignant potential of cells isolated from lymph nodes or brain metastases of melanoma patients and implications for prognosis. Cancer Res. 1991;51:2029. [PubMed] [Google Scholar]

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