[Molecular biological and immunohistochemical analysis of tp53 in human ameloblastomas]. / Molekularbiologische und immunhistochemische Untersuchung des tp53-Gens in menschlichen Ameloblastomen.

AIM: Ameloblastoma is the most frequent epithelial tumor of the jaws. The spread is locally invasive and it tends to recur. Malignant transformation and occurrence of metastases has been described. Immunohistochemical analysis shows an enhanced expression of P53 protein in ameloblastomas. Mutation of the tp53 tumor suppressor gene was verified in several human tumors. In this study histological sections were analyzed for the expression of P53 protein and the tp53 gene was examined for mutations. MATERIAL AND METHODS: Tumor DNA from 29 patients with an ameloblastoma was examined for mutations in exons 5 to 8 of the tp53 tumor suppressor gene using PCR, SSCP,- and sequential analysis. Histological sections of the tumors were analyzed immunohistochemically for an overexpression of P53 protein. RESULTS: Two tp53 mutations (6.9%) in ameloblastomas were verified for the first time. In all 58.6% of the tumors showed an immunoreactivity for P53 protein. There was a statistically significant positive correlation (Fisher's exact test p<0.0148) between an increased number of P53-positive tumor cells and the appearance of recurrence. DISCUSSION: In the face of the uncertain postoperative behavior of ameloblastomas, the immunohistochemical verification of more than 10% P53-positive tumor cells may give a prognostic indication for a tendency to recurrence.

Analysis of the RET, GDNF, EDN3, and EDNRB genes in patients with intestinal neuronal dysplasia and Hirschsprung disease.

BACKGROUND: Hirschsprung disease (HSCR) is a frequent congenital disorder with an incidence of 1 in 5000 live births, characterised by the absence of parasympathetic intramural ganglion cells in the hindgut resulting in intestinal obstruction in neonates and severe constipation in infants and adults. Intestinal neuronal dysplasia (IND) shares clinical features with HSCR but the submucosal parasympathetic plexus is affected. IND has been proposed as one of the most frequent causes of chronic constipation and is often associated with HSCR. METHODS: We examined 29 patients diagnosed with sporadic HSCR, 20 patients with IND, and 12 patients with mixed HSCR/IND for mutations in the coding regions of the RET, GDNF, EDNRB, and EDN3 genes. The entire coding regions were analysed by single strand conformational polymorphism and DNA sequencing. RESULTS: Only three RET mutations were detected in patients with HSCR. In patients with IND or a mixed HSCR/IND phenotype, no mutations in these genes were observed. While HSCR and HSCR/IND showed over representation of a specific RET polymorphism in exon 2, IND exhibited a significantly lower frequency comparable with that of controls. CONCLUSIONS: The mutation frequency found in our sporadic HSCR patients (10%) and the allelic distribution of RET polymorphisms are comparable with earlier published data. A significantly different allelic distribution in an established HSCR associated polymorphism argues against common genetic pathways for HSCR and IND.