Abstract
Pheochromocytomas are catecholamine-producing tumors of the adult adrenal medulla. They are rare in humans and most other species but common in laboratory rats. However, the relevance of rat pheochromocytomas as a model for their human counterparts is uncertain. Previous studies of spontaneous and drug-induced rat pheochromocytomas and the PC12 pheochromocytoma cell line suggested a distinctive noradrenergic phenotype, possibly reflecting origin from a progenitor not present in the adult human adrenal. In this study, we studied 31 pheochromocytomas derived from test and control male and female rats in toxicologic studies for expression of the epinephrine-synthesizing enzyme phenylethanolamine-N-methyltransferase (PNMT) and the receptor tyrosine kinase Ret. PNMT, which defines adrenergic chromaffin cells, is frequently expressed in human pheochromocytomas, often in tumors that also overexpress RET. We also tested for the expression of the cell cycle checkpoint protein p27Kip1, which recently was reported absent in pheochromocytomas from a strain of rats with a hereditary mixed multiple endocrine neoplasia (MEN)-like syndrome. Using immunoblots, we demonstrated PNMT expression in almost 50% of the 31 tumors, although often at lower levels than in normal rat adrenal medulla. The majority of tumors overexpressed Ret. There was no apparent correlation between PNMT and Ret. However, in this study, PNMT expression was strongly associated with tumors arising in female rats, while overexpression of Ret did not show a sex predilection. Robust expression of p27Kip1 was seen in all tumors from the toxicologic studies and also in a small sample of pheochromocytomas from Long–Evans rats, which were reported to have a mixed MEN-like syndrome in the 1980s. The present results show that rat pheochromocytomas have greater phenotypic diversity than previously believed and greater similarity to their human counterparts with respect to these two important markers. Loss of p27Kip1 does not appear to account for the high frequency of pheochromocytomas in commonly utilized rat strains.
Similar content being viewed by others
References
Dahia PL. Evolving concepts in pheochromocytoma and paraganglioma. Curr Opin Oncol 18:1–8, 2006.
Brouwers FM, Glasker S, Nave AF, et al. Proteomic profiling of von Hippel–Lindau syndrome and multiple endocrine neoplasia type 2 pheochromocytomas reveals different expression of chromogranin B. Endocr Relat Cancer 14:463–71, 2007.
Eisenhofer G, Huynh TT, Pacak K, et al. Distinct gene expression profiles in norepinephrine- and epinephrine-producing hereditary and sporadic pheochromocytomas: activation of hypoxia-driven angiogenic pathways in von Hippel–Lindau syndrome. Endocr Relat Cancer 11:897–911, 2004.
Tischler AS. Molecular and cellular biology of pheochromocytomas and extra-adrenal paragangliomas. Endocr Pathol 17:321–8, 2006.
Wada M, Asai N, Tsuzuki T, et al. Detection of ret homodimers in MEN 2A-associated pheochromocytomas. Biochem Biophys Res Commun 218:606–9, 1996.
Takaya K, Yoshimasa T, Arai H, et al. The RET proto-oncogene in sporadic pheochromocytomas. Intern Med. 35:449–52, 1996.
Tischler AS, Powers JF, Alroy J. Animal models of pheochromocytoma. Histol Histopathol 19:883–95, 2004.
Eranko O. Nodular hyperplasia and increase of noradrenaline content in the adrenal medulla of nicotine-treated rats. Acta Pathol Microbiol Scand 36:210–8, 1955.
Tischler AS, Coupland RE. Changes in structure and function of the adrenal medulla. In: Mohr U, Dungworth CC, Capen CC, eds. Pathobiology of the aging rat. vol 2. Washington, DC: ILSI, pp. 244–68, 1994.
Tischler AS, DeLellis RA, Perlman RL, et al. Spontaneous proliferative lesions of the adrenal medulla in aging Long–Evans rats. Comparison to PC12 cells, small granule-containing cells, and human adrenal medullary hyperplasia. Lab Invest 53:486–98, 1985.
Greene LA, Tischler AS. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci USA 73:2424–8, 1976.
Powers JF, Schelling K, Brachold JM, et al. High-level expression of receptor tyrosine kinase ret and responsiveness to ret-activating ligands in pheochromocytoma cell lines from neurofibromatosis knockout mice. Mol Cell Neurosci 20:382–9, 2002.
Elkahloun AG, Powers JF, Nyska A, Eisenhofer G, Tischler AS. Gene expression profiling of rat pheochromocytoma. Ann NY Acad Sci 1073:290–9, 2006.
Wong D. Why is the adrenal adrenergic? Endocr Pathol 14:25–36, 2003.
Pellegata NS, Quintanilla-Martinez L, Siggelkow H, et al. Germ-line mutations in p27Kip1 cause a multiple endocrine neoplasia syndrome in rats and humans. Proc Natl Acad Sci USA 103:15558–63, 2006.
Lee AK, DeLellis RA, Blount M, Nunnemacher G, Wolfe HJ. Pituitary proliferative lesions in aging male Long–Evans rats. A model of mixed multiple endocrine neoplasia syndrome. Lab Invest 47:595–602, 1982.
Powers JF, Brachold JM, Ehsani SA, Tischler AS. Up-regulation of ret by reserpine in the adult rat adrenal medulla. Neuroscience 132:605–12, 2005.
Powers JF, Evinger MJ, Tsokas P, et al. Pheochromocytoma cell lines from heterozygous neurofibromatosis knockout mice. Cell Tissue Res 302:309–20, 2000.
Le Hir H, Charlet-Berguerand N, Gimenez-Roqueplo A, et al. Relative expression of the RET9 and RET51 isoforms in human pheochromocytomas. Oncology 58:311–8, 2000.
Tsui-Pierchala BA, Ahrens RC, Crowder RJ, Milbrandt J, Johnson EM Jr. The long and short isoforms of Ret function as independent signaling complexes. J Biol Chem 277:34618–25, 2002.
Reznik G, Germann P-G. Ganglioneuroma, adrenal, rat. In: Jones TC, Capen CC, Mohr U, eds. Endocrine system. 2nd ed. Berlin: Springer, pp. 427–32, 1996.
Tischler AS, Powers JF, Pignatello M, Tsokas P, Downing JC, McClain RM. Vitamin D3-induced proliferative lesions in the rat adrenal medulla. Toxicol Sci 51:9–18, 1999.
Warren S, Chute RN. Pheochromocytoma. Cancer 29:327–31, 1972.
Pachnis V, Mankoo B, Costantini F. Expression of the c-ret proto-oncogene during mouse embryogenesis. Development 119:1005–17, 1993.
Tischler AS, Greene LA. Phenotypic plasticity of pheochromocytoma and normal adrenal medullary cells. Adv Biochem Psychopharmacol 25:61–8, 1980.
Olson L. Fluorescence histochemical evidence for axonal growth and secretion from transplanted adrenal medullary tissue. Histochemie 22:1–7, 1970.
Marek L, Levresse V, Amura C, et al. Multiple signaling conduits regulate global differentiation-specific gene expression in PC12 cells. J Cell Physiol 201:459–69, 2004.
Acknowledgments
The authors thank Dr. Robert Maronpot from the NIEHS and Dr. Mel Hamlin from the NTP Archives for their support in providing the samples for this investigation. This research was supported by NIH grants R01 CA48107 and R01 NS37685 (to AST).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Powers, J.F., Picard, K.L., Nyska, A. et al. Adrenergic Differentiation and Ret Expression in Rat Pheochromocytomas. Endocr Pathol 19, 9–16 (2008). https://doi.org/10.1007/s12022-008-9019-1
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12022-008-9019-1