title: Effect of dominant negative transforming growth factor-beta receptor type II on cytotoxic activity of RAW 264.7, a murine macrophage cell line.
abstract: Transforming growth factor-beta (TGF-beta) is a potent suppressor of the immune system. In the present study, we investigated the effect of TGF-beta resistance on a murine macrophage cell line, RAW 264.7, by overexpressing a dominant negative TGF-beta receptor type II (TbetaRIIDN) construct. As expected, TbetaRIIDN-expressing RAW cells, designated as RAW-TbetaRIIDN, were resistant to TGF-beta signaling. When these cells were cocultured with the murine renal cell carcinoma cell line, Renca, a dramatic increase in apoptosis of Renca cells was observed. Simultaneously, elevated levels of inducible nitric oxide synthase (iNOS) and tumor necrosis factor-alpha (TNF-alpha) in association with IFN-gamma were detected in RAW-TbetaRIIDN cells. When the effects of TNF-alpha and iNOS were neutralized through the use of neutralizing antibody and N(G)-methyl-L-arginine, respectively, the enhanced cytotoxicity of TbetaRIIDN-RAW cells was partially reversed. Taken together, these results show that TGF-beta-resistant RAW 264.7 murine macrophage cells have increased cytotoxic activity that is in part mediated by iNOS and TNF-alpha.
authors.affiliation: Division of Urologic Oncology, The Cancer Institute of New Jersey, Robert Wood Johnson Medical School, 195 Little Albany Street, New Brunswick, NJ 08901, USA.
authors.forename: Geun Taek
authors.initials: GT
authors.lastname: Lee
authors.forename: Jun Hyuk
authors.initials: JH
authors.lastname: Hong
authors.forename: Cheol
authors.initials: C
authors.lastname: Kwak
authors.forename: Jaesung
authors.initials: J
authors.lastname: Woo
authors.forename: Victoria
authors.initials: V
authors.lastname: Liu
authors.forename: Chung
authors.initials: C
authors.lastname: Lee
authors.forename: Isaac Yi
authors.initials: IY
authors.lastname: Kim
mesh_terms: D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005799:Genes, Dominant; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases; D000077294:Receptor, Transforming Growth Factor-beta Type II; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics* / Q000502:physiology*; D015398:Signal Transduction; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D018125:Receptors, Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D052247:Nitric Oxide Synthase Type II; D017346:Protein Serine-Threonine Kinases; D000077294:Receptor, Transforming Growth Factor-beta Type II