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Karin Flicker,
Peter Ulz,
Harald Höger,
Petra Zeitlhofer,
Oskar A Haas, Annemarie Behmel,
Wolfgang Buchinger,
Christian Scheuba,
Bruno Niederle,
Roswitha Pfragner,
Michael R Speicher
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ABSTRACT: Hereditary and sporadic medullary thyroid carcinoma (MTC) are closely associated with RET proto-oncogene mutations. However, the role of additional changes in the tumor genomes remains unclear. Our objective was the identification of chromosomal regions involved in MTC tumorigenesis and to assess their significance by using MTC-derived cell lines. We used array-CGH (comparative genomic hybridization) to map chromosomal imbalances in 52 primary tumors and ten metastases. Eleven tumors (11/52, 21%) were hereditary and 41 (41/52, 79%) were sporadic. Among the latter, 15 tumors (15/41, 37%) harbored RET mutations. Furthermore, we characterized five MTC cell lines in detail and evaluated the tumorigenicity by severe combined immunodeficiency (SCID)-mouse experiments. Most MTCs had only few copy number changes, and losses of chromosomes 1p, 4q, 19p and 22q were observed most frequently. The number of chromosomal aberrations increased in metastases. Twenty-three percent (12/52) of the primary tumors did not even show any chromosomal gains and losses. We injected three cell lines (two of these were without chromosomal changes and pathogenic RET mutations) into immune deficient SCID mice, and in each case, we observed rapid tumor growth at the injection sites. Our data suggest that MTCs--in contrast to most other tumor entities--do not acquire a multitude of genomic imbalances. SCID mouse experiments performed with chromosomally normal cell lines and without RET mutations suggest that presently unknown submicroscopic genomic changes are sufficient in MTC tumorigenesis.
International Journal of Cancer 10/2011; 131(2):E66-73. · 5.44 Impact Factor
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Roswitha Pfragner, Annemarie Behmel,
Harald Höger,
Alfred Beham,
Elisabeth Ingolic,
Ingeborg Stelzer,
Bernhard Svejda,
Victor Aguiriano Moser,
Anna Christina Obenauf,
Veronika Siegl,
Oskar Haas,
Bruno Niederle
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ABSTRACT: Carcinoids are rare tumors derived from enterochromaffin (EC) cells of the embryonic neural crest. They have malignant potential and their incidence is steadily increasing. The only curative treatment option is surgery. We have focused on cultivation of human neuroendocrine tumors (NET) as relevant models for the study of potential therapy. Only a few cell lines from human carcinoids have been established so far, among them our earlier KRJ-I cell line from a human ileal carcinoid. The reason for the poor success in establishing carcinoid cell lines is due to the small amount of tissue available and the low mitotic activity in primary cultures. We have successfully established three continuously growing cell lines from tissue obtained from a metastatic human carcinoid of the terminal ileum (midgut carcinoid): P-STS was derived from the primary tumor, L-STS from a lymph node metastasis and H-STS from a hepatic metastasis. Immunocytochemistry proved the maintenance of characteristic neuroendocrine properties. Electron microscopy confirmed the presence of neuroendocrine granules. The three cell lines were tumorigenous in SCID-mice. Cytogenetic analyses revealed clonal tetraploidy, inversion and deletion in chromosome 18q, and non-clonal numerical and structural aberrations. Array CGH did not show notable imbalances. Mutation screening of P-STS excluded a MEN1-gene-associated genetic predisposition with high probability. The novel cell lines P-STS, L-STS and H-STS may be useful in vitro and in vivo models for further studies of biological characteristics and the development of new therapeutic agents.
Anticancer research 07/2009; 29(6):1951-61. · 1.73 Impact Factor
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01/2005: pages 373 - 403; , ISBN: 9780471722786
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ABSTRACT: A unilateral, apparently sporadic pheochromocytoma was removed from the right adrenal of a 73-yr-old Caucasian woman. At the time of surgery, germline DNA from the patient was not available. However, a continuous cell line (KNA) established from the tumor showed a heterozygous sequence variant TGC (cysteine) to TGG (tryptophan) in exon 10, codon 611 of the RET proto-oncogene. Subsequent genetic testing of the patient and her offspring revealed the same base-change in herself, one daughter, one son, and the only grandson, confirming hereditary disease classified as MEN2A-2. Clinical follow up of the patient revealed elevated serum calcitonin after 6 yr. Thyroidectomy was performed and revealed a small medullary thyroid carcinoma. The patient's children thus far show no evidence of MEN2, but C-cell hyperplasia has been diagnosed in the grandson. Our serendipitous finding of a MEN2A-2 mutation in a patient with initial diagnosis of late onset, unilateral, "sporadic" pheochromocytoma would argue for routine mutation screening of even elderly patients presenting with a pheochromocytoma.
Endocrine Pathology 02/2003; 14(4):375-82. · 1.36 Impact Factor
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ABSTRACT: A novel polymerase chain reaction (PCR) technique has been combined with chromosome flow sorting to characterise two lymphoblastoid cell lines and one medullary thyroid carcinoma cell line carrying translocations close to the locus for multiple endocrine neoplasia type 2A (MEN 2A). Five hundred copies of the derivative chromosome(s) were flow sorted from each cell line and amplified by degenerate oligonucleotide-primed–polymerase chain reaction (DOP-PCR). This generated pools of DNA sequences corresponding to the abnormal chromosomes, which were then used as probes in fluorescence in situ hybridisation (FISH) experiments on normal metaphase cells. The resultant chromosome paints revealed the portions of the normal chromosomes related to those involved in the translocations. By this technique, translocation breakpoints in bands p15, q11.2, and q21 of chromosome 10 were defined in the above cell lines, in two cases refining previous cytogenetic data. This study shows that flow sorting of aberrant chromosomes and chromosome painting can be used as a rapid aid to cytogenetic analysis, particularly in cases of difficult karyotypes, such as tumours. Furthermore, the DOP-PCR technique described here will have applications to other areas of genome analysis, such as cloning of new markers; its design will allow a general and representative amplification to occur from any starting DNA in any species.
Genes Chromosomes and Cancer 03/1992; 4(3):257 - 263. · 3.31 Impact Factor
