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Immunohistochemical determination of protein kinase C expression and proliferative activity in human brain tumors

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Summary

Protein kinase C (PKC), the major receptor for phorbol ester tumor promotors, is a phospholipid- and calcium-dependent phosphorylating enzyme which plays an important role in the intracellular signal transduction necessary for a variety of basic cellular functions including the control of cell proliferation. To determine the expression of PKC in human neurogenic tumors we investigated 121 tumors of the human nervous system by means of immunohistochemistry using the monoclonal antibody C5. The results were compared with immunohistochemical staining for intermediate filament proteins, desmoplakins, and the proliferation-associated nuclear antigen Ki-67. Besides strong staining of normal and reactive astrocytes, C5 immunoreactivity was consistently observed in tumor cells of all types of gliomas. However, the fraction of C5 positive tumor cells varied between the different tumor types with astrocytomas and subependymomas demonstrating the strongest immunoreactivity. In the other gliomas, especially those of higher malignancy, a considerable heterogeneity in C5 expression could be observed. There was a tendency for the percentage of C5 immunostained tumor cells being lower in high-grade gliomas compared to low-grade ones and comparison with Ki-67 staining frequently revealed an inverse relationship between proliferative activity and C5 immunoreactivity. Besides the gliomas we found 3 of 7 neurinomas and 6 of 18 meningiomas which were partially C5 positive. All other tumors investigated including medulloblastomas and metastatic carcinomas were C5 negative. Our results thus indicate that immunohistochemistry for PKC using the monoclonal antibody C5 could be an useful aid for histopathological tumor classification in neurooncology.

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References

  1. Berridge MJ (1987) Inositol triphosphate and diacylglycerol: two interacting second messengers. Annu Rev Biochem 56:159–193

    Google Scholar 

  2. Blumberg PM (1988) Protein kinase C as the receptor for the phorbol ester tumor promotors: Sixth Rhoads Memorial Award Lecture. Cancer Res 48:1–8

    Google Scholar 

  3. Brandt SJ, Niedel JE, Bell RM, Young WS (1987) Distinct patterns of expression of different protein kinase C mRNAs in rat tissues. Cell 49:57–63

    Google Scholar 

  4. Cordell JL, Falini B, Erber WN, Ghosh AK, Abdulaziz Z, Macdonald S, Pulford KAF, Stein H, Mason DY (1984) Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes). J Histochem Cytochem 32:219–229

    Google Scholar 

  5. Coussens L, Parker PJ, Rhee L, Yang-Feng TL, Chen E, Waterfield MD, Francke U, Ullrich A (1986) Multiple, distinct forms of bovine and human protein kinase C suggest diversity in cellular signaling pathways. Science 233:859–866

    Google Scholar 

  6. Deck JHN, Rubinstein LJ (1981) Glial fibrillary acidic protein in stromal cells of some capillary hemangioblastomas: significance and possible implications of an immunoperoxidase study. Acta Neuropathol (Berl) 54:173–181

    Google Scholar 

  7. Deckert M, Reifenberger G, Wechsler W (1989) Determination of the proliferative potential of human brain tumors by the monoclonal antibody Ki-67. J Cancer Res Clin Oncol 115:179–188

    Google Scholar 

  8. Fleischman LF, Chahwala SB, Cantley L (1986) Ras-transformed cells: altered levels of phosphatidylinositol-4,5-biphosphate and catabolites. Science 231:407–410

    Google Scholar 

  9. Gerdes J, Schwab U, Lemke H, Stein H (1983) Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation. Int J Cancer 31:13–20

    Google Scholar 

  10. Gerdes J, Lemke H, Baisch H, Wacker HH, Schwab U, Stein H (1984) Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67. J Immunol 133:1710–1715

    Google Scholar 

  11. Girard PR, Mazzai GJ, Wood JG, Kuo JF (1985) Polyclonal antibodies to phospholipid/Ca2+-dependent protein kinase and immunocytochemical localization of the enzyme in rat brain. Proc Natl Acad Sci USA 82:3030–3034

    Google Scholar 

  12. Guillem JG, O'Brien CA, Fitzer CJ, Forde KA, Logerfo P, Treat M, Weinstein IB (1987) Altered levels of protein kinase C and Ca2+-dependent protein kinases in human colon carcinomas. Cancer Res 47:2036–2039

    Google Scholar 

  13. Housey GM, Johnson MD, Hsiao WLW, O'Brian CA, Murphy JP, Kirschmeier P, Weinstein IB (1988) Overproduction of protein kinase C causes disordered growth control in rat fibroblasts. Cell 52:343–354

    Google Scholar 

  14. Hsu SM, Raine L, Fanger H (1981) The use of anti-avidin antibody and avidin-biotin-peroxidase-complex in immunoperoxidase techniques. Am J Clin Pathol 75:816–821

    Google Scholar 

  15. Huang K-P, Huang FL (1986) Immunochemical characterization of rat brain protein kinase C. J Biol Chem 261:14781–14787

    Google Scholar 

  16. Huang K-P, Nakabayashi H, Huang FL (1986) Isozymic forms of rat brain Ca2+-activated and phospholipid-dependent protein kinase. Proc Natl Acad Sci USA 83:8535–8539

    Google Scholar 

  17. Jaken S, Kiley SC (1987) Purification and characterization of three types of protein kinase C from rabbit brain cytosol. Proc Natl Acad Sci USA 84:4418–4422

    Google Scholar 

  18. Kitano T, Hashimoto T, Kikkawa U, Ase K, Saito N, Tanaka C, Ichimori Y, Tsukamoto K, Nishizuka Y (1987) Monoclonal antibodies against rat brain protein kinase C and their application to immunohistochemistry in nervous tissues. J Neurosci 7:1520–1525

    Google Scholar 

  19. Kleihues P, Kiessling M, Janzer RC (1987) Morphological markers in neuro-oncology. Curr Top Pathol 77:307–338

    Google Scholar 

  20. Kuo JF, Andersson RGG, Wise BC, Mackerlova L, Salomonsson I, Brackett NL, Katoh N, Shoji M, Wrenn RW (1980) Calcium-dependent protein kinase: widespread occurrence in various tissues and phyla of the animal kingdom and comparison of effects of phospholipid, calmodulin, and trifluoperazine. Proc Natl Acad Sci USA 77:7039–7043

    Google Scholar 

  21. Lacal JC, Fleming TP, Warren BS, Blumberg PM, Aronson SA (1987) Involvement of functional protein kinase C in the mitogenic response to the H-ras oncogene product. Mol Cell Biol 7:4146–4149

    Google Scholar 

  22. Libermann TA, Razon N, Bartal AD, Yarden Y, Schlessinger J, Soreq H (1984) Expression of epidermal growth factor receptors in human brain tumors. Cancer Res 44:753–760

    Google Scholar 

  23. Libermann TA, Nusbaum HR, Razon N, Kris R, Lax I, Soreq H, Whittles N, Waterfield MD, Ullrich A, Schlessinger J (1985) Amplification, enhanced expression and possible rearrangement of EGF receptor gene in primary human brain tumors of glial origin. Nature 313:144–147

    Google Scholar 

  24. Mochly-Rosen D, Basbaum AI, Koshland DE (1987) Distinct cellular and regional localization of immunoreactive protein kinase C in rat brain. Proc Natl Acad Sci USA 84:4660–4664

    Google Scholar 

  25. Neary JT, Norenberg LB, Norenberg MD (1986) Calcium-activated, phospholipid-dependent protein kinase and protein substrates in primary cultures of astrocytes. Brain Res 385:420–424

    Google Scholar 

  26. Nister M, Libermann TA, Betsholtz C, Pettersson M, Claesson-Welsh L, Heldin CH, Schlessinger J, Westermark B (1988) Expression of messenger RNAs for platelet-derived growth factor and transforming growth factor alpha and their receptors in human malignant glioma cell lines. Cancer Res 48:3910–3918

    Google Scholar 

  27. Nishizuka Y (1986) Studies and perspectives of protein kinase C. Science 233:305–312

    Google Scholar 

  28. Parker PJ, Coussens L, Totty N, Rhee L, Young S, Chen E, Stabel S, Waterfield MD, Ullrich A (1986) The complete primary structure of protein kinase C: the major phorbol ester receptor. Science 233:853–859

    Google Scholar 

  29. Pearce B, Morrow C, Murphy S (1986) Receptor-mediated inositol phospholipid hydrolysis in astrocytes. Eur J Pharmacol 121:231–243

    Google Scholar 

  30. Perentes E, Rubinstein LJ (1987) Recent applications of immunoperoxidase histochemistry in human neuro-oncology. An update. Arch Pathol Lab Med 111:796–812

    Google Scholar 

  31. Persons DA, Wilkison WO, Bell RM, Finn OJ (1988) Altered growth regulation and enhanced tumorigenicity of NIH 3T3 fibroblasts transfected with protein kinase C-I cDNA. Cell 52:447–458

    Google Scholar 

  32. Reifenberger G, Szymaś J, Wechsler W (1987) Differential expression of glial- and neuronal-associated antigens in human tumors of the central and peripheral nervous system. Acta Neuropathol (Berl) 74:105–123

    Google Scholar 

  33. Reifenberger G, Prior R, Deckert M, Wechsler W (1989) Epidermal growth factor receptor expression and growth fraction in human tumours of the nervous system. Virchows Arch [A] 414:147–155

    Google Scholar 

  34. Takai Y, Kishimoto A, Inoue M, Nishizuka Y (1977) Studies on a cyclic nucleotide-independent protein kinase and its proenzyme in mammalian tissues. J Biol Chem 252:7603–7609

    Google Scholar 

  35. Vartanian T, Szuchet S, Dawson G, Campagnoni AT (1986) Oligodendrocyte adhesion activates protein kinase C-mediated phosphorylation of myelin basic protein. Science 234:1395–1398

    Google Scholar 

  36. Winkler J, Roosen N, Reifenberger G, Wechsler W (1988) Dedifferentiation in recurrent astrocytoma: clinicopathological study of two cases. Clin Neuropathol 7:222 [abstr]

    Google Scholar 

  37. Wong AJ, Bigner SH, Bigner DD, Kinzler KW, Hamilton SR, Vogelstein B (1987) Increased expression of the EGF receptor gene in malignant gliomas is invariably associated with gene amplification. Proc Natl Acad Sci USA 84:6899–6903

    Google Scholar 

  38. Woodgett JR, Hunter T (1987) Immunological evidence for two physiological forms of protein kinase C. Mol Cell Biol 7:85–96

    Google Scholar 

  39. Worley PF, Baraban JM, Snyder SH (1986) Heterogenous localization of protein kinase C in rat brain: autoradiographic analysis of phorbol ester receptor binding. J Neurosci 6:199–207

    Google Scholar 

  40. Worley PF, Baraban JM, De Souza EB, Snyder SH (1986) Mapping second messenger systems in the brain: differential localizations of adenylate cyclase and protein kinase C. Proc Natl Acad Sci USA 83:4053–4057

    Google Scholar 

  41. Young S, Rothbard J, Parker PJ (1988) A monoclonal antibody recognizing the site of limited proteolysis of protein kinase C. Eur J Biochem 173:247–252

    Google Scholar 

  42. Zülch KJ (1979) Histological typing of tumours of the central nervous system. International histological classification of tumours. WHO Geneva

    Google Scholar 

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Authors and Affiliations

  1. Abteilung für Neuropathologie, Universität Düsseldorf, Moorenstrasse 5, D-4000, Düsseldorf 1, Federal Republic of Germany

    G. Reifenberger, M. Deckert & W. Wechsler

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  1. G. Reifenberger
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  2. M. Deckert
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  3. W. Wechsler
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Supported by the Deutsche Forschungsgemeinschaft, SFB 200

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Reifenberger, G., Deckert, M. & Wechsler, W. Immunohistochemical determination of protein kinase C expression and proliferative activity in human brain tumors. Acta Neuropathol 78, 166–175 (1989). https://doi.org/10.1007/BF00688205

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  • DOI: https://doi.org/10.1007/BF00688205

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