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Rapid estimation of the proliferating index of brain tumours

  • Laboratory Investigation
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Summary

A rapid and versatile method has been developed for estimating proliferating/labelling indices (enumerated S-phase cells) of brain tumours. Fragments of fresh, surgically excised tumour specimens are labelled in vitro with 10 µM bromodeoxyuridine (BrdU) at 37° for 1 hour in MEM + 10% FCS, washed in PBS, blotted dry and then imprinted on glass slides leaving a mirror image monolayer of cells of the impressed surface. The incorporated BrdU is then detected indirectly by immunoperoxidase staining after fixation.

No fewer than 1000 cells are counted under high power and the PI is expressed as the % of BrdU positive cells to the total number counted. The PI of the various tumours analysed ranged 2.54–13.08% for gliomas, 5.29 – 7.60% for metastases, 0 – 0.83% for benign meningiomas. Thirty one cases have been studied.

Results, obtainable within a few hours of tumour biopsy/resection, correlate favourably with published flowcytometry proliferating indices for the tumours analysed and thus validate the method as a comparable alternative to in vivo BrdU labelling.

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References

  1. Hoshino T, Wilson CB: Cell kinetic analyses of human malignant brain tumours (gliomas). Cancer 44: 956–962, 1979

    Google Scholar 

  2. Hoshino T, Kobayashi S, Townsend JJ, Wilson CB: A cell kinetic study on medulloblastomas. Cancer 55: 1711–1713, 1985

    Google Scholar 

  3. Hoshino T, Nagashima T, Murovic JA, Wilson CB, Edwards MSB, Gutin PH, Davis RL, Dearmond SJ: In situ cell kinetics studies on human neuroectodermal tumours with bromodeoxymidine labelling. J Neurosurg 64: 453–459, 1986

    Google Scholar 

  4. Gerdes J, Schwab U, Lempke H, Stein H: An immunohistochemical method for estimating cell growth fractions in rapid histological diagnosis during surgery. Int J Cancer 31: 13–20, 1983

    CAS  PubMed  Google Scholar 

  5. Landolt AM, Shibata T, Kleihues P: Growth rate of human pituitary adenomas. J Neurosurg 67: 803–806, 1987

    Google Scholar 

  6. Gerdes J, Lemke H, Baisch H, Wacker H, Schwab U, Stein H: Cell cycle analysis of a cell proliferation associated human nuclear antigen defined by the monoclonal antibody Ki67. J Immunol 133: 1710–1715, 1984

    Google Scholar 

  7. Lloyd RV, Wilson BS, Varani J, Gaur PK, Moline S, Makari JG: Immunocytochemical characterization of a monoclonal antibody that recognises mitosing cells. Am J Pathol 121:275–283, 1985

    Google Scholar 

  8. Robbins BA, Vega D de la, Ogata K, Tan EM, Nakamura RM: Immunohistochemical detection proliferating cell nuclear antigen in solid human malignancies. Arch Path Lab Med 111: 841–845, 1987

    Google Scholar 

  9. Gerdes J: An immunohistochemical method for estimating cell growth fractions in rapid histological diagnosis during surgery. Int J Cancer 35: 169–171, 1985

    Google Scholar 

  10. Persson H, Hennighausen L, Taub R, DeGrado W, Leder P: Antibodies to human c-myconcogene product: Evidence of an evolutionarily conserved protein induced during cell proliferation. Science 225: 687–694, 1984

    Google Scholar 

  11. Dippold WG, Jay G, Delves A, Khoury G, Old LJ: p53 transformation related protein: Detection by monoclonal antibody in mouse and human cells. Proc Nat Acad Sci USA 8: 1695–1699, 1981

    Google Scholar 

  12. Szybalski W: X-ray sensitization by halopyrimidine. Cancer Chemother Rep 58: 539–557, 1974

    Google Scholar 

  13. Gratzner HG: Monoclonal antibody to 5-bromo and 5-iododeoxyuridine: a new reagent for detection of DNA replication. Science 218: 474–475, 1982

    Google Scholar 

  14. Nagashima T, DeArmond SJ, Murovik JA, Hoshino T: Immunocytochemical demonstration of S-phase cells by antibromodeoxyuridine monoclonal antibody in human brain tumour tissues. Acta Neuropathol 67: 155–159, 1985

    Google Scholar 

  15. Nagashima T, Hoshino T, Cho KG, Senegor M, Waldman F, Nomura K: Comparison of bromodeoxyuridine labelling indices obtained from tissue sections and flow cytometry of brain tumours. J Neurosurg 68: 388–392, 1988

    Google Scholar 

  16. Chigasaki H: Studies on the DNA synthesis function of glial cells by means of 3H-thymidine microradiography. Brain Nerve 15: 767–781, 1963

    Google Scholar 

  17. Johnson HA, Haymaker WE, Rubini JR, Fliedner TM, Bond VP, Cronkite EP, Hughes WL: A radioautographic study of a human brain and glioblastoma multiforme after the invivo uptake of tritiated thymidine. Cancer 13: 636–642, 1960

    Google Scholar 

  18. Hoshino T, Barker M, Wilson CB, Boldrey EB, Fewer D: Cell kinetics of human gliomas. J Neurosurg 37: 15–26, 1972

    Google Scholar 

  19. Hoshino T, Wilson CB, Rosenblum ML, Barker M: Chemotherapeutic implication of growth fraction and cell cycle time in glioblastoma. J Neurosurg 43: 127–135, 1975

    Google Scholar 

  20. Van Dilla MA, Trujille TT, Mullanuy PE, Coulter JR: Cell microfluorometry: A method for rapid fluorescence measurement. Science 163: 1213–1214, 1969

    Google Scholar 

  21. Hitchcock E, Morris CS, Sotelo MG, Salmon M: A comparison of smear and imprint techniques for rapid diagnosis in neuro-oncology. Surg Neurol 26:176–182, 1986

    Google Scholar 

  22. Kyung CG, Hoshino T, Nagashima T, Murovic JA, Wilson CB: Prediction of tumour doubling time in recurrent meningiomas. J Neurosurg 65: 790–794, 1986

    Google Scholar 

  23. Hoshino T, Nagashima T, Cho K, Murovic J, Hodes J, Wilson C, Edwards M, Pitts L: S-phase fraction of human brain tumours insitu measured by uptake of bromdeoxyuridine. Int J Cancer 38: 369–374, 1986

    Google Scholar 

  24. Goz B: The effects of incorporation of 5-halogenated deoxyuridines into the DNA of eukaryotic cells. Pharmacol Rev 29:249–272, 1978

    Google Scholar 

  25. Ellwart J, Dormer P: Effect of 5-fluoro-2-deoxyuridine (FdUrd) on 5-bromo-2-deoxyuridine (BrdUrd) incorporation into DNA measured with a monoclonal BrdUrd antibody and by the BrdUrd/Hoechst quenching effect. Cytometry 6: 513–520, 1985

    Google Scholar 

  26. Yoshii Y, Narushima K, Tsuboi K, Maki Y, Sugiyama K: Tumour cord and growth in human brain tumours based on mathematical morphology. J Neuro-oncology 6: 119–128, 1988

    Google Scholar 

  27. Nishizaki T, Orita T, Saiki M, Furutani Y, Aoki H: Cell kinetics studies of human brain tumours by invitro labelling using anti-BUdR monoclonal antibody. J Neurosurg 69: 371–374, 1988

    Google Scholar 

  28. Morimura T, Kitz K, Budka H: In situ analysis of cell kinetics in human brain tumours. Acta Neuropathol 77: 276–282, 1989

    Google Scholar 

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Authors and Affiliations

  1. University Brain Tumour Laboratory, Department of Neurosurgery, University of Birmingham, Midland Centre for Neurosurgery and Neurology, B67 7JX, Smethwick, West Midlands, United Kingdom

    A. Detta & E. Hitchcock

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  1. A. Detta
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  2. E. Hitchcock
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Detta, A., Hitchcock, E. Rapid estimation of the proliferating index of brain tumours. J Neuro-Oncol 8, 245–253 (1990). https://doi.org/10.1007/BF00177357

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