RESUMO
The WT1 transcription factor regulates SRY expression during the initial steps of the sex determination process in humans, activating a gene cascade leading to testis differentiation. In addition to causing Wilms' tumor, mutations in WT1 are often responsible for urogenital defects in men, while SRY mutations are mainly related to 46,XY pure gonadal dysgenesis. In order to evaluate their role in abnormal testicular organogenesis, we screened for SRY and WT1 gene mutations in 10 children with XY partial gonadal dysgenesis, 2 of whom with a history of Wilms' tumor. The open reading frame and 360 bp of the 5' flanking sequence of the SRY gene, and the ten exons and intron boundaries of the WT1 gene were amplified by PCR of genomic DNA. Single-strand conformation polymorphism was initially used for WT1 mutation screening. Since shifts in fragment migration were only observed for intron/exon 4, the ten WT1 exons from all patients were sequenced manually. No mutations were detected in the SRY 5' untranslated region or within SRY open-reading frame sequences. WT1 sequencing revealed one missense mutation (D396N) in the ninth exon of a patient who also had Wilms' tumor. In addition, two silent point mutations were found in the first exon including one described here for the first time. Some non-coding sequence variations were detected, representing one new (IVS4+85A>G) and two already described (-7ATG T>G, IVS9-49 T>C) single nucleotide polymorphisms. Therefore, mutations in two major genes required for gonadal development, SRY and WT1, are not responsible for XY partial gonadal dysgenesis.
Assuntos
Proteínas de Ligação a DNA/genética , Genes do Tumor de Wilms , Disgenesia Gonadal 46 XY/genética , Mutação/genética , Proteínas Nucleares/genética , Testículo/embriologia , Fatores de Transcrição/genética , Regiões 5' não Traduzidas/genética , Sequência de Bases , Criança , Pré-Escolar , Éxons , Humanos , Lactente , Masculino , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Fenótipo , Reação em Cadeia da Polimerase , Proteína da Região Y Determinante do SexoRESUMO
The WT1 transcription factor regulates SRY expression during the initial steps of the sex determination process in humans, activating a gene cascade leading to testis differentiation. In addition to causing Wilms' tumor, mutations in WT1 are often responsible for urogenital defects in men, while SRY mutations are mainly related to 46,XY pure gonadal dysgenesis. In order to evaluate their role in abnormal testicular organogenesis, we screened for SRY and WT1 gene mutations in 10 children with XY partial gonadal dysgenesis, 2 of whom with a history of Wilms' tumor. The open reading frame and 360 bp of the 5' flanking sequence of the SRY gene, and the ten exons and intron boundaries of the WT1 gene were amplified by PCR of genomic DNA. Single-strand conformation polymorphism was initially used for WT1 mutation screening. Since shifts in fragment migration were only observed for intron/exon 4, the ten WT1 exons from all patients were sequenced manually. No mutations were detected in the SRY 5' untranslated region or within SRY open-reading frame sequences. WT1 sequencing revealed one missense mutation (D396N) in the ninth exon of a patient who also had Wilms' tumor. In addition, two silent point mutations were found in the first exon including one described here for the first time. Some non-coding sequence variations were detected, representing one new (IVS4+85A>G) and two already described (-7ATG T>G, IVS9-49 T>C) single nucleotide polymorphisms. Therefore, mutations in two major genes required for gonadal development, SRY and WT1, are not responsible for XY partial gonadal dysgenesis.
Assuntos
Humanos , Masculino , Lactente , Pré-Escolar , Criança , Proteínas de Ligação a DNA/genética , Genes do Tumor de Wilms , /genética , Mutação/genética , Testículo/embriologia , /genética , Sequência de Bases , Éxons , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Fenótipo , Reação em Cadeia da PolimeraseRESUMO
UNLABELLED: The aim was to correlate histological findings in cervix lesions to human papillomavirus (HPV), as detected by polymerase chain reaction (PCR). One hundred and seven women with atypical Pap smear were submitted to colposcopic examination, and suspicious images were biopsied. The PCR assay was performed with the primers MY09/11 and GP05/06+ and, as control, the beta-globin gene was amplified. The morphological findings were correlated to HPV positivity: parakeratosis, acanthosis, koilocytotic atypia (KA), binucleation, dyskeratosis, and number of mitoses. From 107 patients, 61 biopsies were taken: 11 chronic cervicitis (CC), 36 cervical intraepithelial neoplasia (CIN) (13 CIN I; 10 CIN II; 13 CIN III), and 14 suggestive for HPV (SHPV). DNA extraction was not possible in eight cases. HPV was found in 35% CC, 77% CIN, and 64% SHPV. The analysis did not indicate any morphological criteria strongly related to HPV. The findings with highest sensitivity for HPV were KA (88.89%) and binucleation (75%), but with low specificity of 29.41 and 52.94%, respectively. The higher predictive positive values (PV+) for HPV were also KA (72.73%) and binucleation (77.14%). Considering KA, dyskeratosis and binucleation together, PV+ was 72.41%. CONCLUSION: Although indicative, none of the studied morphological criteria was always related to PCR virus detection, denoting some limitations for histological diagnosis.
