title: Carcinoembryonic antigen-stimulated THP-1 macrophages activate endothelial cells and increase cell-cell adhesion of colorectal cancer cells.
abstract: The liver is the most common site for metastasis by colorectal cancer, and numerous studies have shown a relationship between serum carcinoembryonic antigen (CEA) levels and metastasis to this site. CEA activates hepatic macrophages or Kupffer cells via binding to the CEA receptor (CEA-R), which results in the production of cytokines and the up-regulation of endothelial adhesion molecules, both of which are implicated in hepatic metastasis. Since tissue macrophages implicated in the metastatic process can often be difficult to isolate, the aim of this study was to develop an in vitro model system to study the complex mechanisms of CEA-induced macrophage activation and metastasis. Undifferentiated, human monocytic THP-1 (U-THP) cells were differentiated (D-THP) to macrophages by exposure to 200 ng/ml phorbol myristate acetate (PMA) for 18 h. Immunohistochemistry showed two CEA-R isoforms present in both U- and D-THP cells. The receptors were localized primarily to the nucleus in U-THP cells, while a significant cell-surface presence was observed following PMA-differentiation. Incubation of D-THP-1 cells with CEA resulted in a significant increase in tumor necrosis factor-alpha (TNF-alpha) release over 24 h compared to untreated D-THP-1 or U-THP controls confirming the functionality of these cell surface receptors. U-THP cells were unresponsive to CEA. Attachment of HT-29 cells to human umbilical vein endothelial cells significantly increased at 1 h after incubation with both recombinant TNF-alpha and conditioned media from CEA stimulated D-THP cells by six and eightfold, respectively. This study establishes an in vitro system utilizing a human macrophage cell line expressing functional CEA-Rs to study activation and signaling mechanisms of CEA that facilitate tumor cell attachment to activated endothelial cells. Utilization of this in vitro system may lead to a more complete understanding of the expression and function of CEA-R and facilitate the design of anti-CEA-R therapeutic modalities that may significantly diminish the metastatic potential of CEA overexpressing colorectal tumors.
authors.affiliation: Department of Surgery, Boston University School of Medicine, Boston, MA 02118, USA. astucchi@bu.edu
authors.forename: Cary B
authors.initials: CB
authors.lastname: Aarons
authors.forename: Olga
authors.initials: O
authors.lastname: Bajenova
authors.forename: Charles
authors.initials: C
authors.lastname: Andrews
authors.forename: Stanley
authors.initials: S
authors.lastname: Heydrick
authors.forename: Kristen N
authors.initials: KN
authors.lastname: Bushell
authors.forename: Karen L
authors.initials: KL
authors.lastname: Reed
authors.forename: Peter
authors.initials: P
authors.lastname: Thomas
authors.forename: James M
authors.initials: JM
authors.lastname: Becker
authors.forename: Arthur F
authors.initials: AF
authors.lastname: Stucchi
mesh_terms: D002272:Carcinoembryonic Antigen / Q000502:physiology*; D002448:Cell Adhesion; D002460:Cell Line; D015179:Colorectal Neoplasms / Q000473:pathology*; D019040:E-Selectin / Q000235:genetics; D042783:Endothelial Cells / Q000502:physiology*; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics; D008264:Macrophages / Q000502:physiology*; D011956:Receptors, Cell Surface / Q000502:physiology; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D002272:Carcinoembryonic Antigen; D019040:E-Selectin; D011956:Receptors, Cell Surface; D014409:Tumor Necrosis Factor-alpha; C065686:carcinoembryonic antigen binding protein, human; D018799:Intercellular Adhesion Molecule-1