Aberrant methylation patterns of the Rassf1a gene in rat lung adenocarcinomas induced by N-nitrosobis(2-hydroxypropyl)amine
| Autor: | Kanya Honoki, Mayumi Shigemura, Hiromasa Fujii, Ayumi Kato, Toshifumi Tsujiuchi, Kyoko Shimizu |
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| Rok vydání: | 2005 |
| Předmět: |
Gene isoform
Male endocrine system Cancer Research Lung Neoplasms Nitrosamines Bisulfite sequencing Molecular Sequence Data Biology Adenocarcinoma medicine.disease_cause Exon Reference Values medicine Animals Molecular Biology Lung Base Sequence Tumor Suppressor Proteins Single-strand conformation polymorphism Methylation DNA Methylation medicine.disease Molecular biology Rats DNA methylation Mutation Carcinogenesis |
| Zdroj: | Molecular carcinogenesis. 45(2) |
| ISSN: | 0899-1987 |
| Popis: | To clarify the involvement of the Rassf1a gene in lung carcinogenesis, we investigated the methylation status in the 5' upstream region of the RAS-association domain family 1, isoform A (Rassf1a) gene in rat lung adenocarcinomas induced by N-nitrosobis(2-hydroxypropyl)amine (BHP). Six-week-old male Wistar rats were given 2000 ppm BHP in their drinking water for 12 wk and maintained without further treatment until they were sacrificed at 25 wk. A total of 15 lung adenocarcinomas were obtained and total RNAs were extracted from each for assessment of expression of the Rassf1a gene by reverse transcription (RT)-polymerase chain reaction (PCR) analysis. To measure the methylation status of the Rassf1a gene, five adenocarcinomas with a marked reduction of the Rassf1a expression and two normal lung tissues were used for a bisulfite sequencing analysis. While the normal lung tissue was unmethylated, all five adenocarcinomas were highly methylated in the 5' upstream region. Genomic DNAs were also extracted from 15 adenocarcinomas, and mutation analysis of the Rassf1a was performed with PCR-single-strand conformation polymorphism (SSCP) analysis. No mutations were detected throughout exons 1-6. The present results suggest that the aberrant methylation may be involved in the inactivation of the Rassf1a gene in the development of lung adenocarcinomas induced by BHP in rats. |
| Databáze: | OpenAIRE |
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