Genes targeted by the estrogen and progesterone receptors in the human endometrial cell lines HEC1A and RL95-2.
BACKGROUND
When the steroid hormones estrogen and progesterone bind to nuclear receptors, they have transcriptional impact on target genes in the human endometrium. These transcriptional changes have a critical function in preparing the endometrium for embryo implantation.


METHODS
382 genes were selected, differentially expressed in the receptive endometrium, to study their responsiveness of estrogen and progesterone. The endometrial cell lines HEC1A and RL95-2 were used as experimental models for the non-receptive and receptive endometrium, respectively. Putative targets for activated steroid hormone receptors were investigated by chromatin immunoprecipitation (ChIP) using receptor-specific antibodies. Promoter occupancy of the selected genes by steroid receptors was detected in ChIP-purified DNA by quantitative PCR (qPCR). Expression analysis by reverse transcriptase (RT)-PCR was used to further investigate hormone dependent mRNA expression regulation of a subset of genes.


RESULTS
ChIP-qPCR analysis demonstrated that each steroid hormone receptor had distinct group of target genes in the endometrial cell lines. After estradiol treatment, expression of estrogen receptor target genes predominated in HEC1A cells (n = 137) compared to RL95-2 cells (n = 35). In contrast, expression of progesterone receptor target genes was higher in RL95-2 cells (n = 83) than in HEC1A cells (n = 7) after progesterone treatment. RT-PCR analysis of 20 genes demonstrated transcriptional changes after estradiol or progesterone treatment of the cell lines.


CONCLUSIONS
Combined results from ChIP-qPCR and RT-PCR analysis showed different patterns of steroid hormone receptor occupancy at target genes, corresponding to activation or suppression of gene expression after hormone treatment of HEC1A and RL95-2 cell lines.
Tamm
Karin
K
Centre for Biology of Integrated Systems, Tallinn University of Technology, Tallinn, Estonia. karin.tamm@mail.ee
Rõõm
Miia
M
Salumets
Andres
A
Metsis
Madis
M
D002460:Cell Line; D002843:Chromatin / Q000737:chemistry / Q000235:genetics; D003627:Data Interpretation, Statistical; D004717:Endometrium / Q000378:metabolism*; D005260:Female; D018390:Gene Targeting; D006801:Humans; D047468:Immunoprecipitation; D066298:In Vitro Techniques; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011960:Receptors, Estrogen / Q000235:genetics*; D011980:Receptors, Progesterone / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
D002843:Chromatin; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone