Allelic loss of chromosomes 8 and 19 in MENX-associated rat pheochromocytoma
Alena Shyla
Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Institute of Radiation Biology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorGabriele Hölzlwimmer
Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorJulia Calzada-Wack
Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorKarin Bink
Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorOleg Tischenko
Institute of Radiation Protection, Medical Physics Group, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorMarie-Noëlle Guilly
CEA, DSV, IRCM, SREIT, Laboratoire de Cancérologie Expérimentale, BP6, France
Search for more papers by this authorSylvie Chevillard
CEA, DSV, IRCM, SREIT, Laboratoire de Cancérologie Expérimentale, BP6, France
Search for more papers by this authorElenore Samson
Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorJochen Graw
Institute of Developmental Genetics, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorMichael J. Atkinson
Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Institute of Radiation Biology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorCorresponding Author
Natalia S. Pellegata
Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Fax: +49-89-3187-3360.
Helmholtz Zentrum München-German Research Center for Environment and Health, Institute of Pathology, Ingolstädter Landstraβe 1, D-85764 Neuherberg, GermanySearch for more papers by this authorAlena Shyla
Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Institute of Radiation Biology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorGabriele Hölzlwimmer
Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorJulia Calzada-Wack
Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorKarin Bink
Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorOleg Tischenko
Institute of Radiation Protection, Medical Physics Group, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorMarie-Noëlle Guilly
CEA, DSV, IRCM, SREIT, Laboratoire de Cancérologie Expérimentale, BP6, France
Search for more papers by this authorSylvie Chevillard
CEA, DSV, IRCM, SREIT, Laboratoire de Cancérologie Expérimentale, BP6, France
Search for more papers by this authorElenore Samson
Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorJochen Graw
Institute of Developmental Genetics, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorMichael J. Atkinson
Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Institute of Radiation Biology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Search for more papers by this authorCorresponding Author
Natalia S. Pellegata
Institute of Pathology, Helmholtz Zentrum München-German Research Center for Environment and Health, Neuherberg, Germany
Fax: +49-89-3187-3360.
Helmholtz Zentrum München-German Research Center for Environment and Health, Institute of Pathology, Ingolstädter Landstraβe 1, D-85764 Neuherberg, GermanySearch for more papers by this authorAbstract
Pheochromocytomas are neoplasias of neural crest origin that arise from the chromaffin cells of the adrenal medulla. Pheochromocytomas arise with complete penetrance in rats homozygous for a germ-line frameshift mutation of Cdkn1b, encoding the cell cycle inhibitor p27KIP1 (MENX syndrome). We performed a genome-wide scan for allelic imbalance comparing 20 rat pheochromocytoma DNAs with normal rat DNA to better understand the pathobiology of the tumors and to correlate the findings with human pheochromocytoma. We identified allelic imbalance (AI) at candidate regions on rat chromosomes 8 and 19. Interestingly, the regions often lost in rat tumors are syntenic to regions involved in human pheochromocytomas. Fluorescence in situ hybridization analysis further validated the AI data. Sdhd and Rassf1a were analyzed in detail as they map to regions of AI on chromosome 8 and their homologues are implicated in human pheochromocytoma: we found no genetic mutations nor decreased expression. We also analyzed additional candidate genes, that is, rat homologues of genes predisposing to human pheochromocytoma and known tumor-suppressor genes, but we found no AI. In contrast, we observed frequent overexpression of Cdkn2a and Cdkn2c, encoding the cell cycle inhibitors p16INK4a and p18INK4c, respectively. The relative small number of allelic changes we found in rat pheochromocytoma might be related to their nonmalignant status and losses at chromosomes 8 and 19 are events that precede malignancy. Because of the high concordance of affected loci between rat and human tumors, studies of the MENX-associated pheochromocytomas should facilitate the identification of novel candidate genes implicated in their human counterpart.
Supporting Information
Additional Supporting Information may be found in the online version of this article.
| Filename | Description |
|---|---|
| IJC_24925_sm_suppfig1.tif6 MB | Supporting Figure 1 Real-time RT-PCR shows no change in the expression of Sdhd and Rassf1a in adrenal glands of MENX-affected and control rats. (a) The average level of Sdhd mRNA in normal medullary tissue of adrenal glands of wild-type animals (n = 4) was arbitrarily set to 1 (Controls). Values are the mean of two experiments ± standard deviation. “MENX-PCs” corresponds to the mean expression value of Sdhd in all the various pheochromocytoma samples. There is no significant difference in relative amount of mRNA between the analysed pheochromocytoma cases and the control group (p = 0.33; two-sided t-test). (b) The average level of Rassf1a mRNA in pheochromocytoma samples and normal medullary tissue was estimated as reported in (a). There is no significant difference in relative amount of mRNA between the analysed pheochromocytoma cases and the control group (p = 0.79; two-sided t-test). Cases PC13 and PC20 marked in red both have an allelic imbalance on chromosome 8 where Sdhd maps to 8q23 and Rassf1a maps to 8q32. |
| IJC_24925_sm_suppfig2.tif503.6 KB | Supporting Figure 2 Real-time RT-PCR shows significant increased expression of the tyrosine hydroxylase (Th) gene in macrodissected adrenomedullary tissue (med) compared to adrenocortical tissue (cor). The level of Th mRNA in normal adrenomedullary tissue of wild-type animals (n=5; med 1-5) is shown in blue and in a pooled mRNA derived from 3 independent adrenocortical tissue samples is shown in red (cor). Values are the mean of two experiments ± standard deviation. |
| IJC_24925_sm_suppinfo.doc23.5 KB | Supporting Information. |
| IJC_24925_sm_supptables.doc8.8 MB | Supporting Tables. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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