Skip to main content
NCBI home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1990 Dec;62(6):925–931. doi: 10.1038/bjc.1990.411

Immunohistochemical detection of a hypoxia marker in spontaneous canine tumours.

J M Cline 1, D E Thrall 1, R L Page 1, A J Franko 1, J A Raleigh 1
PMCID: PMC1971560  PMID: 1701659

Abstract

An immunoperoxidase technique has been used to detect the in vivo binding of a 2-nitroimidazole hypoxia marker in histochemical sections of a variety of excised canine tumours. The binding occurred 10-12 cell diameters away from tumour blood vessels, consistent with the expected location of hypoxic cells in tissues in which oxygen concentration gradients are established by diffusion. Hypoxic fractions ranging from 4 to 13% have been estimated on the basis of morphometric analysis of multiple tumour sections. The binding of the marker was restricted to the cytoplasm of the cells. The marker appeared in regions adjacent to necrosis but also in regions free of necrosis. As in earlier autoradiography studies, binding was occasionally observed in cells adjacent to tumour blood vessels. Generally, binding to normal tissues was not observed. However, binding to smooth muscle cells surrounding arterioles in some sections of normal tissue and tumour tissue was observed.

Full text

PDF
925

Images in this article

926Figure 1
on p.926
927Figure 3
on p.927
927Figure 4
on p.927
927Figure 5
on p.927
928Figure 6
on p.928
928Figure 7
on p.928

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allalunis M. J., Chapman J. D., Turner A. R. Identification of a hypoxic population of bone marrow cells. Int J Radiat Oncol Biol Phys. 1983 Feb;9(2):227–232. doi: 10.1016/0360-3016(83)90104-9. [DOI] [PubMed] [Google Scholar]
  2. Awwad H. K., el Naggar M., Mocktar N., Barsoum M. Intercapillary distance measurement as an indicator of hypoxia in carcinoma of the cervix uteri. Int J Radiat Oncol Biol Phys. 1986 Aug;12(8):1329–1333. doi: 10.1016/0360-3016(86)90165-3. [DOI] [PubMed] [Google Scholar]
  3. Brown J. M. Evidence for acutely hypoxic cells in mouse tumours, and a possible mechanism of reoxygenation. Br J Radiol. 1979 Aug;52(620):650–656. doi: 10.1259/0007-1285-52-620-650. [DOI] [PubMed] [Google Scholar]
  4. Brunmark A., Cadenas E., Segura-Aguilar J., Lind C., Ernster L. DT-diaphorase-catalyzed two-electron reduction of various p-benzoquinone- and 1,4-naphthoquinone epoxides. Free Radic Biol Med. 1988;5(3):133–143. doi: 10.1016/0891-5849(88)90076-7. [DOI] [PubMed] [Google Scholar]
  5. Bryant D. W., McCalla D. R. Nitrofuran induced mutagenesis and error prone repair in Escherichia coli. Chem Biol Interact. 1980 Aug;31(2):151–166. doi: 10.1016/0009-2797(80)90002-2. [DOI] [PubMed] [Google Scholar]
  6. Bush R. S., Jenkin R. D., Allt W. E., Beale F. A., Bean H., Dembo A. J., Pringle J. F. Definitive evidence for hypoxic cells influencing cure in cancer therapy. Br J Cancer Suppl. 1978 Jun;3:302–306. [PMC free article] [PubMed] [Google Scholar]
  7. Chacon E., Morrow C. J., Leon A. A., Born J. L., Smith B. R. Regioselective formation of a misonidazole-glutathione conjugate as a function of pH during chemical reduction. Biochem Pharmacol. 1988 Jan 15;37(2):361–363. doi: 10.1016/0006-2952(88)90743-5. [DOI] [PubMed] [Google Scholar]
  8. Chapman J. D., Franko A. J., Sharplin J. A marker for hypoxic cells in tumours with potential clinical applicability. Br J Cancer. 1981 Apr;43(4):546–550. doi: 10.1038/bjc.1981.79. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chapman J. D. Hypoxic sensitizers--implications for radiation therapy. N Engl J Med. 1979 Dec 27;301(26):1429–1432. doi: 10.1056/NEJM197912273012606. [DOI] [PubMed] [Google Scholar]
  10. Cobb L. M., Hacker T., Nolan J. NAD(P)H nitroblue tetrazolium reductase levels in apparently normoxic tissues: a histochemical study correlating enzyme activity with binding of radiolabelled misonidazole. Br J Cancer. 1990 Apr;61(4):524–529. doi: 10.1038/bjc.1990.118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Creasey W. A., Thrall D. E. Pharmacokinetic and anti-tumor studies with the radiosensitizer misonidazole in dogs with spontaneous fibrosarcomas. Am J Vet Res. 1982 Jun;43(6):1015–1018. [PubMed] [Google Scholar]
  12. Franko A. J., Koch C. J. Binding of misonidazole to V79 spheroids and fragments of Dunning rat prostatic and human colon carcinomas in vitro: diffusion of oxygen and reactive metabolites. Int J Radiat Oncol Biol Phys. 1984 Aug;10(8):1333–1336. doi: 10.1016/0360-3016(84)90343-2. [DOI] [PubMed] [Google Scholar]
  13. Franko A. J., Koch C. J., Garrecht B. M., Sharplin J., Hughes D. Oxygen dependence of binding of misonidazole to rodent and human tumors in vitro. Cancer Res. 1987 Oct 15;47(20):5367–5376. [PubMed] [Google Scholar]
  14. Franko A. J. Misonidazole and other hypoxia markers: metabolism and applications. Int J Radiat Oncol Biol Phys. 1986 Jul;12(7):1195–1202. doi: 10.1016/0360-3016(86)90257-9. [DOI] [PubMed] [Google Scholar]
  15. Franko A. J., Raleigh J. A., Sutherland R. G., Soderlind K. J. Metabolic binding of misonidazole to mouse tissues. Comparison between labels on the ring and side chain, and the production of tritiated water. Biochem Pharmacol. 1989 Feb 15;38(4):665–670. doi: 10.1016/0006-2952(89)90213-x. [DOI] [PubMed] [Google Scholar]
  16. Franko A. J., Sutherland R. M. Oxygen diffusion distance and development of necrosis in multicell spheroids. Radiat Res. 1979 Sep;79(3):439–453. [PubMed] [Google Scholar]
  17. Garcia C. F., Weiss L. M., Lowder J., Komoroske C., Link M. P., Levy R., Warnke R. A. Quantitation and estimation of lymphocyte subsets in tissue sections. Comparison with flow cytometry. Am J Clin Pathol. 1987 Apr;87(4):470–477. doi: 10.1093/ajcp/87.4.470. [DOI] [PubMed] [Google Scholar]
  18. Gatenby R. A., Kessler H. B., Rosenblum J. S., Coia L. R., Moldofsky P. J., Hartz W. H., Broder G. J. Oxygen distribution in squamous cell carcinoma metastases and its relationship to outcome of radiation therapy. Int J Radiat Oncol Biol Phys. 1988 May;14(5):831–838. doi: 10.1016/0360-3016(88)90002-8. [DOI] [PubMed] [Google Scholar]
  19. Hendry J. H. Quantitation of the radiotherapeutic importance of naturally-hypoxic normal tissues from collated experiments with rodents using single doses. Int J Radiat Oncol Biol Phys. 1979 Jul;5(7):971–976. doi: 10.1016/0360-3016(79)90602-3. [DOI] [PubMed] [Google Scholar]
  20. Hirst D. G., Denekamp J., Hobson B. Proliferation kinetics of endothelial and tumour cells in three mouse mammary carcinomas. Cell Tissue Kinet. 1982 May;15(3):251–261. doi: 10.1111/j.1365-2184.1982.tb01044.x. [DOI] [PubMed] [Google Scholar]
  21. Jette D. C., Wiebe L. I., Chapman J. D. Synthesis and in vivo studies of the radiosensitizer 4-[82Br]bromomisonidazole. Int J Nucl Med Biol. 1983;10(4):205–210. doi: 10.1016/0047-0740(83)90080-3. [DOI] [PubMed] [Google Scholar]
  22. Kallman R. F. The phenomenon of reoxygenation and its implications for fractionated radiotherapy. Radiology. 1972 Oct;105(1):135–142. doi: 10.1148/105.1.135. [DOI] [PubMed] [Google Scholar]
  23. Kennedy K. A. Hypoxic cells as specific drug targets for chemotherapy. Anticancer Drug Des. 1987 Oct;2(2):181–194. [PubMed] [Google Scholar]
  24. Maxwell R. J., Workman P., Griffiths J. R. Demonstration of tumor-selective retention of fluorinated nitroimidazole probes by 19F magnetic resonance spectroscopy in vivo. Int J Radiat Oncol Biol Phys. 1989 Apr;16(4):925–929. doi: 10.1016/0360-3016(89)90888-2. [DOI] [PubMed] [Google Scholar]
  25. Miller G. G., Best M. W., Franko A. J., Koch C. J., Raleigh J. A. Quantitation of hypoxia in multicellular spheroids by video image analysis. Int J Radiat Oncol Biol Phys. 1989 Apr;16(4):949–952. doi: 10.1016/0360-3016(89)90893-6. [DOI] [PubMed] [Google Scholar]
  26. Owen L. N., Steel G. G. The growth and cell population kinetics of spontaneous tumours in domestic animals. Br J Cancer. 1969 Sep;23(3):493–509. doi: 10.1038/bjc.1969.62. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Popel A. S., Pittman R. N., Ellsworth M. L. Rate of oxygen loss from arterioles is an order of magnitude higher than expected. Am J Physiol. 1989 Mar;256(3 Pt 2):H921–H924. doi: 10.1152/ajpheart.1989.256.3.H921. [DOI] [PubMed] [Google Scholar]
  28. Raleigh J. A., Franko A. J., Koch C. J., Born J. L. Binding of misonidazole to hypoxic cells in monolayer and spheroid culture: evidence that a side-chain label is bound as efficiently as a ring label. Br J Cancer. 1985 Feb;51(2):229–235. doi: 10.1038/bjc.1985.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Raleigh J. A., Miller G. G., Franko A. J., Koch C. J., Fuciarelli A. F., Kelly D. A. Fluorescence immunohistochemical detection of hypoxic cells in spheroids and tumours. Br J Cancer. 1987 Oct;56(4):395–400. doi: 10.1038/bjc.1987.213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rasey J. S., Koh W. J., Grierson J. R., Grunbaum Z., Krohn K. A. Radiolabelled fluoromisonidazole as an imaging agent for tumor hypoxia. Int J Radiat Oncol Biol Phys. 1989 Nov;17(5):985–991. doi: 10.1016/0360-3016(89)90146-6. [DOI] [PubMed] [Google Scholar]
  31. Rauth A. M., McClelland R. A., Michaels H. B., Battistella R. The oxygen dependence of the reduction of nitroimidazoles in a radiolytic model system. Int J Radiat Oncol Biol Phys. 1984 Aug;10(8):1323–1326. doi: 10.1016/0360-3016(84)90341-9. [DOI] [PubMed] [Google Scholar]
  32. Smith B. R. Hypoxia-enhanced reduction and covalent binding of [2-3H]misonidazole in the perfused rat liver. Biochem Pharmacol. 1984 Apr 15;33(8):1379–1381. doi: 10.1016/0006-2952(84)90199-0. [DOI] [PubMed] [Google Scholar]
  33. THOMLINSON R. H., GRAY L. H. The histological structure of some human lung cancers and the possible implications for radiotherapy. Br J Cancer. 1955 Dec;9(4):539–549. doi: 10.1038/bjc.1955.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tracy J. W., Catto B. A., Webster L. T., Jr Reductive metabolism of niridazole by adult Schistosoma mansoni. Correlation with covalent drug binding to parasite macromolecules. Mol Pharmacol. 1983 Sep;24(2):291–299. [PubMed] [Google Scholar]
  35. Urtasun R. C., Koch C. J., Franko A. J., Raleigh J. A., Chapman J. D. A novel technique for measuring human tissue pO2 at the cellular level. Br J Cancer. 1986 Sep;54(3):453–457. doi: 10.1038/bjc.1986.197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Van Os-Corby D. J., Koch C. J., Chapman J. D. Is misonidazole binding to mouse tissues a measure of cellular pO2? Biochem Pharmacol. 1987 Oct 15;36(20):3487–3494. doi: 10.1016/0006-2952(87)90330-3. [DOI] [PubMed] [Google Scholar]
  37. Vanhoutte P. M. The endothelium--modulator of vascular smooth-muscle tone. N Engl J Med. 1988 Aug 25;319(8):512–513. doi: 10.1056/NEJM198808253190809. [DOI] [PubMed] [Google Scholar]
  38. Varghese A. J. Glutathione conjugates of misonidazole. Biochem Biophys Res Commun. 1983 May 16;112(3):1013–1020. doi: 10.1016/0006-291x(83)91719-9. [DOI] [PubMed] [Google Scholar]
  39. Varghese A. J., Whitmore G. F. Binding to cellular macromolecules as a possible mechanism for the cytotoxicity of misonidazole. Cancer Res. 1980 Jul;40(7):2165–2169. [PubMed] [Google Scholar]
  40. White R., Workman P., Owen L. The pharmacokinetics in mice and dogs of nitroimidazole radiosensitizers and chemosensitizers more lipophilic than misonidazole. Int J Radiat Oncol Biol Phys. 1982 Mar-Apr;8(3-4):473–476. doi: 10.1016/0360-3016(82)90664-2. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES