Demonstration of iNOS-mRNA and iNOS in human monocytes stimulated with cancer cells in vitro
| Autor: | Jerzy Stachura, Marek Zembala, Maciej Siedlar, Irena Ruggiero, Jerzy Więckiewicz, Bozenna Mytar, Mariola Hyszko, Agnieszka Krzeszowiak, Jarosław Baran |
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| Rok vydání: | 1999 |
| Předmět: |
Cytotoxicity
Immunologic Immunology Nitric Oxide Synthase Type II Cell Communication In situ hybridization Adenocarcinoma Nitric Oxide Monocytes Flow cytometry Nitric oxide chemistry.chemical_compound Tumor Cells Cultured medicine Humans Immunology and Allergy Cytotoxic T cell RNA Messenger In Situ Hybridization biology medicine.diagnostic_test Reverse Transcriptase Polymerase Chain Reaction hemic and immune systems Cell Biology Flow Cytometry Molecular biology Coculture Techniques In vitro Nitric oxide synthase chemistry Cell culture Cancer cell biology.protein Nitric Oxide Synthase Colorectal Neoplasms |
| Zdroj: | Journal of Leukocyte Biology. 65:597-604 |
| ISSN: | 1938-3673 0741-5400 |
| DOI: | 10.1002/jlb.65.5.597 |
| Popis: | Synthesis and localization of inducible nitric oxide synthase mRNA (iNOS-mRNA) and iNOS protein in the cultures of human monocytes (Mφ) and colon carcinoma cell line (DeTa) that resulted in nitric oxide (NO) synthesis has been studied. The iNOS-mRNA was observed around the sixth hour of culture and peaked at the twelfth hour. The iNOS-mRNA, as determined by the in situ hybridization and iNOS protein, as detected by staining with specific anti-iNOS monoclonal antibodies, were observed preferentially in the cytoplasm of some Mφ, but not in cancer cells. Mφ cultured alone did not show detectable iNOS-mRNA expression and iNOS protein. Mφ sorted out from tumor cells after 8hof co-culture expressed iNOS protein and iNOS-mRNA, which were not detected in Mφ without previous contact with cancer cells. Prevention of NO synthesis by (L-N5-1-iminoethyl)-ornithine (L-NIO) partly inhibited Mφ cytotoxic activity against DeTa (NO-inducing cancer cell line) but not against the human pancreatic cancer (HPC-4) cell line that does not induce NO production in Mφ. This suggests that Mφ cytotoxic activity, at least in some cases, may be NO dependent. These observations provide further evidence that Mφ can be directly stimulated by cancer cells for de novo production of NO and suggest that iNOS occurring in the tumor-infiltrating macrophages may arise as a result of their interactions with tumor cells. However, because only some tumor cells are able to induce NO production in a small proportion of Mφ, its role in the anti-tumor response of the host is probably limited. J. Leukoc. Biol. 65: 597–604; 1999. |
| Databáze: | OpenAIRE |
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