engleski

Methylation of BRCA1 gene is not involved in mouse teratocarcinoma development

DNA methylation is the important epigenetic mechanism regulating normal embryonal development but also involved in the development of cancer. Experimental mouse teratocarcinoma as a research model that connects embryogenesis and cancerogenesis, two processes often studied separately. It is not yet known why the normal gastrulating mouse embryo transplanted at an ectopic site gives rise to a malignant tumor, the teratocarcinoma. Teratocarcinoma contains embryonal carcinoma cells (EC), the malignant counterpart of embryonic stem (ES) cells. Tumor suppressor gene BRCA1 is one of many genes suspected to be involved in the process of EC cell development. Therefore DNA methylation status of the BRCA1 promoter region was investigated by pyrosequencing assay, recognized as a powerful tool for DNA methylation analyses. 7, 5– days-old C3H embryos were microsugically isolated and spent 4 weeks under the kidney capsule of syngeneic adults. DNA was isolated from embryos and teratocarcinomas. Bisulfite conversion was performed by Methylation-GoldTM Kit, Zymo Research. Converted DNA was amplified in PCR using specific primers for a 200bp fragment of BRCA1 promoter. A fragment of 22bp was analyzed for DNA methylation status in the pyrosequencing machine PSQ 96 MA. Complete demethylation of the BRCA1 gene promoter in both 7, 5-days-old embryo-proper and teratocarcinoma was discovered. It seems that the methylation of BRCA1 is not involved in the development of experimental mouse teratocarcinoma.

BRCA1; teratocarcinoma; DNA methylation

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