Skip to main content
SpringerLink
Log in

First continuous human pheochromocytoma cell line: KNA Biological, cytogenetic and molecular characterization of KNA cells

  • Published:
Journal of Neurocytology

Abstract

Pheochromocytomas are rare tumours, with an incidence of 1–2 per million which arise from chromaffin cells of the adrenal medulla. They occur sporadically or as part of dominantly inherited cancer syndromes like multiple endocrine neoplasia 2 (MEN2A and 2B) and others. Continuous cell lines, not available so far, are essential tools for studies in these tumours. A continuous cell line (KNA) was established from a sporadic pheochromocytoma of the right adrenal gland of a 73-year-old woman. The KNA cells grow as suspensions of spheroids and show the morphological and immunocytochemical characteristics of neuronal chromaffin cells, such as neuroendocrine granules, and positive reactions to chromogranin- and related peptide-, neuron specific enolase and vasoactive intestinal peptide antibodies. Neurite-like processes are formed after addition of nerve growth factor. Chromosomal analyses revealed a diploid (46,XX, n=50) to hypodiploid (43–45,XX, n=15) karyotype. In hypodiploid metaphases most frequently #19, #17, #21 and #22 were missing. Chromosome arms 1p and 4q showed apparently consistent interstitial deletions: 6q, 8q, 13q and 22q showed clonal interstitial deletions. The cell line shows a heterozygous sequence variant TGC (cysteine) to TGG (tryptophan) in codon 611 in exon 10 of the RET proto-oncogene. So far, PC-12, a rat adrenal pheochromocytoma, has been the only continuous pheochromocytoma cell line available. KNA represents the first report on a human continuous pheochromocytoma cell line, the first report of structural chromosome aberrations in pheochromocytomas and the first report of a RET mutation TGC to TGG in exon 10 of the RET proto-oncogene in a sporadic pheochromocytoma. © 1998 Chapman and Hall

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Andersen, L. B., Fountain, J. W., Gutmann, D. H., Tarle, S. A., Glover, T. W., Dracopoli, N. C., Housman, D. E. & Collins, F. S. (1993) Mutations in the neurofibromatosis 1 gene in sporadic malignant melanomas. Nature Genetics 3, 118–21.

    Google Scholar 

  • Arnold, A. (1996) RET mutation screening in sporadic pheochromocytoma. Journal of Clinical Endocrinology and Metabolism 81, 430 (letter).

    Google Scholar 

  • Behmel, A., Reichl, E., Pfragner, R. & Niederle, B. (1997) High incidence of gonosomal aneuploidy in lymphocytes of patients with sporadic, MEN1 and MEN2 associated neuroendocrine tumors: agerelated X chromosome instability versus genuine mosaicism. Medizinische Genetik 1, 44, 1–3.

    Google Scholar 

  • Benedum, U. M., Baeuerle, P. A., Koneckid, S., Frank, R., Rowell, J. & Mallet, J. (1986) The primary structure of bovine chromogranin A: a representative of a class of acidic secretory proteins common to a variety of peptidergic cells. EMBO Journal 5, 1495–502.

    Google Scholar 

  • Benedum, U. M., Lamouroux, A., Konecki, D. S., Rosa, P., Hille, A., Baeuerle, P. A., Frank, R., Lottspicgs, S., Mallet, J. & Huttner, W. B. (1987) The primary structure of human secretogranin I (chromogranin B): comparison with chromogranin A reveals homologous terminal domains and a large intervening variable region. EMBO Journal 6, 1203–11.

    Google Scholar 

  • Brower, M., Carney, D. N., Oie, H. K., Gazdar, A. F. & Minna, I. D. (1986) Growth of cell lines and clinical specimens of human non-small cell lung cancer in serum-free defined medium. Cancer Research 46, 798–806.

    Google Scholar 

  • Caillard, T., Opstal, W. Y., Scarcerieux, V., Billardon, C. & Rostene, W. (1995) Treatment of PC12 cells by nerve growth factor, dexamethasone, and forskolin. Effects on cell morphology and expression of neurotensin and tyrosin hydroxylase. Molecular Neurobiology 10, 105–14.

    Google Scholar 

  • Decker, H. J. H., Walter, T. A., Neumann, H. J. H. & Sandberg, G. A. (1988) Cytogenetics of familial pheochromocytoma: importance of trisomy 7 in tumor progression? Blut 57, 270.

    Google Scholar 

  • Dinges, H. P., Wirnsberger, G. & HÖfler, H. (1989) Immunocytochemistry in cytology. Comparative evaluation of different techniques. Analytical and Quantitative Cytology and Histology 11, 22–32.

    Google Scholar 

  • Eng, C., Mulligan, L. M., Smith, D. P., Healey, C. S., Frilling, A., Raue, F., Neumann, H. P., Ponder, M. A. & Ponder, B. A. J. (1995) Low frequency of germline mutations in the RET protooncogene in patients with apparently sporadic medullary thyroid carcinoma. Clinical Endocrinology 43, 123–7.

    Google Scholar 

  • Eng, C., Clayton, D., Schuffenecker, I., Lenoir, G., Cote, G., Gagel, R. F., Ploos van Amstel, H. K., Lips, C. J. M., Nishisho, I., Takai, S. C., Marsh, D. J., Robinson, B. G., Frank-Raue, K., Raue, F., Xue, F., Noll, W. W., Romei, C., Pacini, F., Fink, M., Niederle, B., Zadenius, J., Nordenskjold, M., Komminoth, P., Handy, G. H., Gharib, H., Thibodeau, S. N., Lacroix, A., Frilling, A., Ponder, B. A. J. & Mulligan, L. M. (1996) The relationship between specific RET proto-oncogene mutations and disease phenotype in Multiple Endocrine Neoplasia Type 2. Journal of American Medical Association 276, 1575–9.

    Google Scholar 

  • Eng, C., Mulligan, L. M., Healey, C. S., Houghton, C., Frilling, A., Raue, F., Thomas, G. A. & Ponder, B. A. J. (1996) Heterogeneous mutation of the RET proto-oncogene in subpopulations of medullary thyroid carcinoma. Cancer Research 56, 2167–70.

    Google Scholar 

  • Eng, C. & Mulligan, L. M. (1997) Mutations of the RET proto-oncogene in the Multiple Endocrine Neoplasia Type 2 syndromes, related sporadic tumours, and Hirschsprung disease. Human Mutation 9, 97–109.

    Google Scholar 

  • Evers, B. M., Rady, P. L., Stephen, K., Tyring, K., Sanchez, R. L., Rajamaran, S., Townsend, M., Jr. & Thompson, J. C. (1992) Amplification of the HER-2/neu proto-oncogene in human endocrine tumors. Surgery 112, 211–18.

    Google Scholar 

  • Geldorf, A. A. (1995) Nerve-growth-factor-dependent neurite outgrowth assay; a ressearch model for chemotherapy-induced neuropathy. Journal of Cancer Research and Clinical Oncology 121, 657–69.

    Google Scholar 

  • Greene, L. A. & Rein, G. (1977a) Release, storage and uptake of catecholamines by a clonal line of nerve growth factor responsive pheochromocytoma cells. Brain Research 129, 247–63.

    Google Scholar 

  • Greene, L. A. & Rein, G. (1977b) Synthesis, storage and release of acetylcholine by a noradrenergic pheochromocytoma cell line. Nature 268, 349.

    Google Scholar 

  • Greene, L. A. & Tischler, A. S. (1976) Establishment of a noradrenergic clonal line of rat pheochromocytoma cells which respond to nerve growth factor. Proceedings of the National Academy of Sciences USA 73, 2424–26.

    Google Scholar 

  • Gutmann, D. H., Cole, J. L., Stone, W. J., Ponder, B. A. J. & Collins, F. S. (1994) Loss of neurofibromin in adrenal gland tumors from patients with neurofibromatosis type I. Genes, Chromosomes & Cancer 10, 55–8.

    Google Scholar 

  • Gutmann, D. H., Geist, R. T., Kamala, R., Wallin, G. & Moley, J. F. (1993) Gene expression in pheochromocytomas from patients without NFI. Genes, Chromosomes & Cancer 13, 104–9.

    Google Scholar 

  • Hoch-Ligeti, C. & Camp, J. L. (1959) Catecholamine production in tissue culture of human adrenal medullary tumors and adrenal medulla. Proceedings of the Society for Experimental Biology and Medicine 102, 692–3.

    Google Scholar 

  • HÖfler, H., PÜtz, B., Ruhri, C., Wirnsberger, G., Klimpfinger, M. & Smolle, J. (1987) Simultaneous localization of calcitonin mRNA and peptide in a medullary thyroid carcinoma. Virchows Archiv 54, 144–51.

    Google Scholar 

  • Iyengar, S., Tallini, G., Sirugo, G., Bale, A. E. & Kidd, K. K. (1995) Mutation of the vHL Gene in individuals with sporadic and familial pheochromocytoma.American Journal of Human Genetics 57, Supplement A67,357.

    Google Scholar 

  • Johnson, M. & Johnson, B. (1984) Efficient synthesis of high specific 35S-labeled human b-globin pre-mRNA. Biotechniques 2, 156–162.

    Google Scholar 

  • Jordan, D. K., Patil, S. R., Divelbiss, J. E., Vemuganti, S., Headley, C., Waziri, H. & Gurll, J. (1989) Cytogenetic abnormalities in tumors of patients with von Hippel-Lindau disease. Cancer Genetics and Cytogenetics 42, 227–41.

    Google Scholar 

  • Khosla, S., Patel, V. M., Hay, I. D., Schaid, D. J., Grant, S., van Heerden, J. A. & Thibodeau, S. N. (1991) Loss of heterozygosity suggests multiple genetic alterations in pheochromocytomas and medullary thyroid carcinomas. Journal of Clinical Investigation 87, 1691–9.

    Google Scholar 

  • Lasfargues, E. Y. & Moore, D. H. (1971) A method for the continuous cultivation of mammary epithelium. In Vitro Cellular & Developmental Biology-Animal 7, 21–25.

    Google Scholar 

  • Luft, J. H. (1961) Improvements in epoxy resin embedding methods. Journal of Biophysical & Biochemical Cytology 9, 409–14.

    Google Scholar 

  • Mathew, C. G. P., Smith, B. A., Thorpe, K., Wong, Z., Royle, N. J., Jeffries, A. J. & Ponder, B. A. J. (1987) Deletion of genes on chromosome 1 in endocrine neoplasia. Nature 328, 524–26.

    Google Scholar 

  • Modigliani, E., Vasen, H. M., Raue, K., Dralle, H., Frilling, A., Gheri, G., Brandi, M. L., Limberg, E., Niederle, B., Forgas, L., Rosenberg-Bourgin, M., Calmettes, C. & The European Study Group (1995) Pheochromocytoma in multiple endocrine neoplasia type 2: European study. Journal of Internal Medicine 238, 363–7.

    Google Scholar 

  • Moley, J. F., Brother, M. B., Fong, C. T., White, P. S., Baylin, S. B., Nelkin, B., Wells, S. A. & Brodeur, G. M. (1992) Consistent association of 1p loss of heterozygosity with pheochromocytomas from patients with multiple endocrine neoplasia type 2 syndromes. Cancer Research 52, 770–4.

    Google Scholar 

  • Mulligan, L. M., Gardner, E., Smith, B. A., Mathew, C. G. & Ponder, B. A. J. (1993) Genetic events in tumour initiation and progression in multiple endocrine neoplasia type 2. Genes Chromosomes & Cancer 6, 166–77.

    Google Scholar 

  • Nagatsu, T. & Udenfriend, S. (1972) Photometric assay of dopamine β-hydroxylase. Clinical Chemistry 18, 980–983.

    Google Scholar 

  • Neumann, H. J. H., Schempp, W. & Wienker, T. F. (1988) High-resolution chromosome banding and fragile site studies in von Hippel-Lindau syndrome. Cancer Genetics Cytogenetics 31, 41–46.

    Google Scholar 

  • Pfragner, R., HÖfler, H., Behmel, A., Ingolic, E. & Walser, V. (1990) Establishment and characterization of continuous cell line MTC-SK derived from a human medullary thyroid carcinoma. Cancer Research 50, 4160–66.

    Google Scholar 

  • Pfragner, R., Sadjak, A. & Walser, V. (1984) The effect of nerve growth factor (NGF) on the catecholamine contents of two human pheochromocytomas in tisue culture. Experimental Pathology 26, 21–31.

    Google Scholar 

  • Pfragner, R. & Walser, V. (1980) Long-term tissue culture of human pheochromocytomas. Experimental Pathology 18, 423–9.

    Google Scholar 

  • Pfragner, R. Wirnsberger, G., Behmel, A., Wolf, G., Passath, A., Ingolic, E., Adamiker, D. & Schauenstein, K. (1993) New continuous cell line from human medullary thyroid carcinoma: SINJ. Phenotypic analysis and in vivo carcinogenesis. International Journal of Oncology 2, 831–36.

    Google Scholar 

  • Pfragner, R., Wirnsberger, G., Niederle, B., Behmel, A., Rinner, I., Mandl, A., Wawrina, F., Luo, J. S., Adamiker, D., HÖger, H., Ingolic, E. & Schauenstein, K. (1996) Establishment of a continuous cell line from a human carcinoid of the small intestine (KRJ-I): Characterization and effects of 5-azacytidine on proliferation. International Journal of Oncology 8, 513–520.

    Google Scholar 

  • Polinger, I. S. (1970) Separation of cell types in embryonic heart cell cultures. Experimental Cell Research 63, 78–82.

    Google Scholar 

  • porta, S., Epple, A., Kvetnansky, R., Leitner, G., Emsenhuber, W. & Radkohl, W. (1995) Temporary increase of plasma epinephrine affects stress response 24 hours later. Physiology & Behaviour 88, 223–8.

    Google Scholar 

  • REUBI, C. Waser, B., Khosla, S., Kvols, L., Geollner, J. R., Krenning, E. & Lamberts, S. (1992) In vitro and in vivo detection of somatostatin receptors in pheochromocytomas and paragangliomas. Journal of Clinical Endocrinology and Metabolism 74, 1082–89

    Google Scholar 

  • Reynolds, E. S. (1963) The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. Journal of Cell Biology 17, 208–12.

    Google Scholar 

  • Rodien, P., Jeunemaitre, X., Dumont, C., Beldjord, C. & Plouin, P. F. (1997) Genetic alterations of the RET proto-onogene in familial and sporadic pheochromocytomas. Hormone Research 47, 263–268.

    Google Scholar 

  • Ryan, U. S. & Hart, M. A. (1986) Electron microscopy of endothelial cells in culture I. Transmission microscopy. Journal of Tissue Culture Methods 10, 31–33.

    Google Scholar 

  • Ryan, U. S. & Hart, M. A. (1986) Electron microscopy II. Scanning electron microscopy and OTOTO impregnation method. Journal of Tissue Culture Methods 10, 35–36.

    Google Scholar 

  • Shine, E., Fujita, S., Takami, K., Kurahashi, H., Kurita, T., Kobayashi, T., Mon, T., Nishisho, I. & Takai, S. (1993) Deletion mapping of chromosome 1p and 22q in pheochromocytoma. Japanese Journal of Cancer Research 84, 402–8.

    Google Scholar 

  • Takai, S., Tataishi, H., Nishisho, I., Miki, T., Motomura, K., Miyauchi, A., Kato, M. & Ikeuchi, T. (1987) Loss of genes on chromosome 22 in medullary thyroid carcinoma and pheochromocytoma. Japanese Journal of Cancer Research 78, 894–98.

    Google Scholar 

  • Tanaka, N., Nishisho, I., Yamamoto, M., Miya, A., Shin, E., Karakawa, K., Fujita, S., Kobayashi, T., Rouleau, G. A., Mori, T. & Takai, S. (1992) Loss of heterozygosity on the long arm of chromosome 22 in pheochromocytoma. Genes, Chromosomes & Cancer 5, 399–403.

    Google Scholar 

  • The, I., Murthy, A. E., Hannigan, G. E., Jacoby, L. B., Menon, A. G., Gusella, J. F. & Bednards, A. (1993) Neurofibromatosis type 1 gene mutations in neuroblastoma. Nature Genetics 3, 62–66.

    Google Scholar 

  • Tischler, A. S., Dichter, M. A., Biales, B., Delellis, R. A. Wolfe, H. (1976) Neuronal properties of cultured human endocrine tumor cells of proposed neural crest origin. Science 192, 902–4.

    Google Scholar 

  • Tischler, A. S. & Greene, L. A. (1975) Nerve growth factor-induced process formation by cultured rat pheochromocytoma cells. Nature 258, 341–2.

    Google Scholar 

  • Tischler, A. S. & Greene, A. (1978) Morphologic and cytochemical properties of a clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Laboratory Investigation 39, 77–89.

    Google Scholar 

  • Wirnsberger, G. H., Becker, H., Ziervogel, F. & HÖfler, H. (1992) Diagnostic immunohistochemistry of neuroblastic tumors. American Journal of Surgical Pathology 16, 49–57.

    Google Scholar 

  • Zeiger, M. A., Berton, Z., Keiser, H., Linehan, W. M. & Gnarra, J. R. (1993) Loss of heterozygosity on the short arm of chromosome 3 in sporadic, von Hieppel-Lindau disease-associated, and familial pheochromocytoma. Genes, Chromosomes & Cancer 13, 151–6.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of General and Experimental Pathology, A-8010, Graz, Austria

    R. Pfragner, I. Rinner & S. Porta

  2. Department of Medical Biology and Human Genetics, A-8010, Graz, Austria

    A. Behmel

  3. Department of Internal Medicine, Medical School, University of Graz, Heinrichstrasse 31, A-8010, Graz, Austria

    G. Wirnsberger

  4. SCCRI, St. Anna Children’s Hospital, Vienna, Austria

    T. Henn

  5. Department of Surgery, Medical School, University of Vienna, Austria

    B. Niederle

  6. CRC Human Cancer Genetics Research Group, University of Cambridge, Box 238, Addenbrooke’s Hospital, UK

    D. P. Smith & B. A. J. Ponder

Authors
  1. R. Pfragner
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. Behmel
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. D. P. Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. B. A. J. Ponder
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. G. Wirnsberger
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. I. Rinner
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. S. Porta
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. T. Henn
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. B. Niederle
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pfragner, R., Behmel, A., Smith, D.P. et al. First continuous human pheochromocytoma cell line: KNA Biological, cytogenetic and molecular characterization of KNA cells. J Neurocytol 27, 175–186 (1998). https://doi.org/10.1023/A:1006959625068

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006959625068

Keywords

Search

Navigation