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Quantitation of Heme Oxygenase (HO-1) Copies in Human Tissues by Competitive RT/PCR

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Free Radical and Antioxidant Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 108))

Abstract

A variety of oxidative stress-inducing agents, such as metals, ultra-violet (UV) light, heme, and hemoglobin have been implicated in the pathogenesis of the inflammatory process. The cellular response to such agents involves the production of a number of soluble mediators including acute phase proteins, eicosanoids, and various cytokines.

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References

  1. Shibahara, S, Muller, R. M, and Taguchi, H (1987) Transcriptional control of rat heme oxygenase by heat shock J Biol. Chem. 262, 12,889–12,892

    PubMed  CAS  Google Scholar 

  2. Mitani, K, Fujita, H, Sassa, S., and Kappas, A (1989) Heat shock induction of heme oxygenase mRNA in human Hep3B hepatoma cells. Biochem Biophys Res Commun. 165, 437–441

    Article  PubMed  CAS  Google Scholar 

  3. Keyse, S M and Tyrrell, R. M. (1989) Heme oxygenase is the major 32-kDa stress protein induced in human skin fibroblasts by UVA radiation, hydrogen peroxide, and sodium arsenite Proc. Natl Acad Sci. USA 86, 99–103

    Article  PubMed  CAS  Google Scholar 

  4. Taketani, S, Kohno, H, Yoshinaga, T, and Tokunaga, R (1989) The human 32-kDa stress protein induced by exposure to arsenite and cadmium ions is heme oxygenase. FEBS Lett 245, 173–176

    Article  PubMed  CAS  Google Scholar 

  5. Abraham, N G, Lin, J. H-C, Schwartzman, M L., Levere, R D, and Shibahara, S (1988) The physiological significance of heme oxygenase. Int. J Biochem. 20, 543–558

    Article  PubMed  CAS  Google Scholar 

  6. Stocker, R, Yamamoto, Y, McDonagh, A F, Glazer, A N, and Ames, B N (1987) Bilirubin is an antioxidant of possible physiological importance. Science 235, 1043–1046

    Article  PubMed  CAS  Google Scholar 

  7. Nath, K A, Balla, G, Vercellotti, G. M, Balla, J, Jacob, H. S., Levitt, M. D., and Rosenberg, M E. (1992) Induction of heme oxygenase is a rapid, protective response in rhabdomyolysis in the rat. J. Clin. Invest. 90, 267–270.

    Article  PubMed  CAS  Google Scholar 

  8. Paller, M S and Jacob, H S (1994) Cytochrome P-450 mediates tissue-damaging hydroxyl radical formation during reoxygenation of the kidney Proc. Natl Acad. Sci USA 91, 7002–7006

    Article  PubMed  CAS  Google Scholar 

  9. Stocker, R. (1990) Induction of haem oxygenase as a defence against oxidative stress. Free Rad Res Comm. 9, 101–112

    Article  CAS  Google Scholar 

  10. McCoubrey, W K, Jr., Ewing, J. F., and Maines, M D (1992) Human heme oxygenase-2: characterization and expression of a full-length cDNA and evidence suggesting that the two HO-2 transcripts may differ by choice of a polyadenylation signal. Arch. Biochem. Biophys 295, 13–20

    Article  PubMed  CAS  Google Scholar 

  11. Shibahara, S, Yoshizawa, M, Suzuki, H, Takeda, K, Meguro, K, and Endo, K (1993) Functional analysis of cDNAs for two types of human heme oxygenase and evidence for their separate regulation J Biochem 113, 214–218

    PubMed  CAS  Google Scholar 

  12. Cantoni, L., Rossi, C, Rizzardini, M., Gadina, M., and Ghezzi, P. (1991) Interleukin-1 and tumour necrosis factor induce hepatic haem oxygenase Biochem J. 279, 891–894.

    PubMed  CAS  Google Scholar 

  13. Rizzardini, M, Terao, M, Falciani, F, and Cantoni, L (1993) Cytokine induction of haem oxygenase mRNA in mouse liver. Biochem. J. 290, 343–347.

    PubMed  CAS  Google Scholar 

  14. Lutton, J. D, da Silva, J-L, Moquattash, S, Brown, A C, Levere, R D, and Abraham, N. G (1992) Differential induction of heme oxygenase in the hepatocarcinoma cell line (Hep3b) by environmental agents. J. Cell Biol. 49, 259–265.

    CAS  Google Scholar 

  15. Alam, J. and Zhining, D (1992) Distal AP-1 binding sites mediate basal level enhancement and TPA induction of the mouse heme oxygenase-1 gene. J. Biol Chem. 267, 21,894–21,900

    PubMed  CAS  Google Scholar 

  16. Lavrovsky, Y., Schwartzman, M. L., Levere, R D, Kappas, A, and Abraham, N G. (1994) Identification of NFkB and AP-2 binding sites in the promoter region of the human heme oxygenase-1 gene. Proc. Natl. Acad. Sci USA 91, 5987–5991.

    Article  PubMed  CAS  Google Scholar 

  17. Becker-Andre, M. and Hahlbrock, K (1989) Absolute mRNA quantification using the polymerase chain reaction (PCR): a novel approach by a PCR aided transcript titration assay (PATTY) Nucleic Acids Res. 17, 9437–9446.

    Article  PubMed  CAS  Google Scholar 

  18. Wang, A M., Doyle, M V., and Mark, D.F. (1989) Quantitation of mRNA by the polymerase chain reaction Proc. Natl Acad. Sci. USA 86, 9717–9721.

    Article  PubMed  CAS  Google Scholar 

  19. Gilliland, G., Perrin, S, Blanchard, K., and Bunn, H. F. (1990) Analysis of cytokines mRNA and DNA: detection and quantitation by competitive polymerase chain reaction. Proc Natl. Acad. Sci USA 87, 2725–2729.

    Article  PubMed  CAS  Google Scholar 

  20. Cross, N. C. (1995) Quantitative PCR techniques and applications. Br J. Haematol. 89, 639–647

    Google Scholar 

  21. Murphy, L. D, Herzog, C. E., Rudick, J. B., Fojo, A. T, and Bates, S E (1990) Biochemistry 29, 10,351–10,356

    Article  PubMed  CAS  Google Scholar 

  22. Kutty, G., Hayden, B., Osawa, Y., Wiggert, B., Chader, G J, and Kutty R K. (1992) Heme oxygenase: expression in human retinal and modulation by stress agents in a human retinoblastoma cell model system. Curr. Eye Res. 11, 153–160

    Article  PubMed  CAS  Google Scholar 

  23. Yoshida, T., Biro, P, Cohen, T., Muller, R. M., and Shibahara, S. (1988) Human heme oxygenase cDNA and induction of its mRNA by hemin. Eur J Biochem. 171, 457–461.

    Article  PubMed  CAS  Google Scholar 

  24. Chomczynski, P. and Sacchi, N. (1987) Single-step method of RNA isolation by acid guanidium. Anal. Biochem. 162, 156–159.

    Article  PubMed  CAS  Google Scholar 

  25. Cross, N C, Feng, L, Chase, A., Bungey, J, Hughes, T P, and Goldman, J. M. (1993) Competitive polymerase chain reaction to estimate the number of BCR-ABL transcripts in chronic myeloid leukemia patients after bone marrow transplantation. Blood 82, 1929–1936.

    PubMed  CAS  Google Scholar 

  26. Frenoy, N., Chabli, A, Sol, O, Goldschmit, E., Lemonnier, M. P., Misset, J L, and Debuire, B. (1994) Application of a new protocol for nested PCR to the detection of minimal residual bcr/abl transcripts. Leukemia 8, 1411–1414

    PubMed  CAS  Google Scholar 

  27. Lion, T., Gaiger, A, Henn, T, Horth, E., Haas, O. A., Geissler K., and Gadner H (1995) Use of quantitative polymerase chain reaction to monitor residual disease in chronic myelogenous leukemia during treatment with interferon Leukemia 9, 1353–1360

    PubMed  CAS  Google Scholar 

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© 1998 Humana Press Inc., Totowa, NJ

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Abraham, N.G. (1998). Quantitation of Heme Oxygenase (HO-1) Copies in Human Tissues by Competitive RT/PCR. In: Armstrong, D. (eds) Free Radical and Antioxidant Protocols. Methods in Molecular Biology™, vol 108. Humana Press. https://doi.org/10.1385/0-89603-472-0:199

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  • DOI: https://doi.org/10.1385/0-89603-472-0:199

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-472-3

  • Online ISBN: 978-1-59259-254-8

  • eBook Packages: Springer Protocols

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