DNA methylation of HSD3B2, NUR77 and RARβ promoter genes is not involved in functional differentiation of human androgen-producing adrenocortical cells

ALONSO BURGOS, MARÍA CECILIA; GOÑI, JAVIER; SARACO, NORA; RIVAROLA, MARCO A.; BELGOROSKY, ALICIA; BAQUEDANO MARÍA SONIA

Paris

Congreso; 55th Annual Meeting of the European Society for Paediatric Endocrinology (ESPE); 2016

Sociedad Europea de Endocrinología Pediátrica (ESPE)

Background: RARb cooperates with Nur77 to in-vitroregulate HSD3B2 transcription. NUR77 expression parallelsHSD3B2 expression with a much lower level in androgen-producingadrenocortical tissues (childhood virilizing adrenocorticaltumours (VAT), fetal zone (FeZ) and zona reticularis, ZR).RARb is down-regulated in starved, hyperandrogenic H295Rcells. However, the mechanisms regulating this expression patternand the relevance of RARb to human adrenal physiology areunknown. Objective: To evaluate developmental changes in DNAmethylation of HSD3B2, NUR77 and RARb genes in VAT andnormal human adrenal tissues (HAT). Method: VAT (n=11, age0.75?4.5 year) and HAT collected from 3postnatal groups: Gr1:6 to 20 year, n=8, post-adrenarche, wereevaluated. Total DNA and RNA from whole tissue and fromlaser capture microdissected zona fasciculata (ZF) and ZR in Gr3were isolated. Total RARb (RARbT) and RARb2 expression werestudied using qRT-PCR. Promoter methylation pattern wereexamined by bisulfite sequencing. Results: RARbT and RARb2mRNAswere similar among HATfrom the 3 groups. RARbT mRNAwas lower in VAT compared to age-matched group HAT (P