272 Evaluation of The Histone H3 Gene Expression and Its Correlation With Transcriptional Activity of Er-Alpha Gene in Women With Endometrial Hyperplasia and Adenocarcinoma

The aim of the study was the estimation of proliferative potential of endometrium with the proliferation marker histone H3 and the evaluation of correlation between H3 gene expression and ER-alfa gene expression in women with endometrial hyperplasia and adenocarcinoma. The material included samples obtained from 84 women: normal proliferative endometrium (20), simple (20), complex (15), atypical hyperplasia (11) and adenocarcinoma (18). Samples from resected uterus were immediately snap-frozen in liquid nitrogen and stored in - 70C until the mRNA molecular assays were performed. Histologic diagnoses were conducted on the same material. Primers and probes sequences for examined genes amplification were designed using the Primer Express software and checked in BLAST database. for all RNA extracts RT-PCR was performed with a sequence detector ABI PRISMTM7700 Taq-Man. Our investigation show that with deepening of the carcinogenesis process the copy number of the H3 was increased. Patients with H3 mRNA copy number higher than 20.000 were always in the endometrium adenocarcinoma group. Negative correlation between histon H3 and ER-alpha expression in reference group was also observed. The estimation of transcriptional activity of histone H3 encoding gene may be used as marker of proliferative potential of endomerium. H3 mRNA copy number lower than 20.000 may be used as the auxiliary indicator for differentiation of atypical hyperplasia and endometrial adenocarcinoma. Negative correlation between histone H3 expression and ER-alpha copy number in proliferative phase may indicate the important role of estrogen alpha receptor in mechanisms that protect the endometrium from hyperplasia.