ER beta regulated transcriptome in endometriotic tissues

The Estrogen Receptor beta (ERβ) has an essential role in endometriosis progression. However, the molecular mechanism of how ERβ drives endometriosis progression is not elucidated, yet. To define the role of genomic ERβ in endometriosis progression, we have employed whole-genome microarray expression profiling as a discovery platform to identify ERβ-regulated transcriptome in endometriotic tissues. To get this transcriptome, we applied endometrium specific ERβ overexpression (ERBOE) mouse model by crossing mouse having a pCAG promoter-loxPSTOPloxP-ERβ cassette with PRCre knockin mice that Cre recombinase cDNA was inserted into exon 1 of PR gene. Endometriosis was surgically induced in ERBOE mice and PRCre mice as the control by transplantation of uterine tissues. The ectopic lesions and eutopic endometrium were harvested at the estrus cycle in 4th weeks after endometriosis induction. ERβ-regulated transcriptome analysis in ectopic lesions: We isolated uteri from ERBOE female mice (eight weeks old) and its female control mice (PRCre/+, eight weeks old) and then made endometrium fragments by using Miltex Medical Biopsy Punch Dermal 2 mm OR Grade. The endometriosis was surgically induced by implanting one uterus fragment isolated from ERBOE female mouse to wild-type intact C57BL/6J mouse (n=9, eight weeks old). We also made endometrial pieces from uteri of female control mice (PRCre/+), and then implanted one endometrial piece to wild-type intact C57BL/6J mouse (n=9, eight weeks old) as the control. At estrus cycle in 4th weeks of endometriosis induction, We harvested ERβ overexpressing and ectopic control lesions and then isolated total RNA from them. After that, we combined total RNA isolated from three ectopic lesions to form a single group. Therefore, we made three RNA groups from nine ERβ overexpressing ectopic lesions and nine ectopic control lesions. ERβ-regulated transcriptome analysis in eutopic endometrium: We isolated uteri from wild-type C57BL/6J female (eight weeks old), and then made endometrium fragments by using Miltex Medical Biopsy Punch Dermal 2 mm OR Grade. The endometriosis was surgically induced by implanting one uterus fragment isolated from wild-type C57BL/6J into ERBOE female mouse (eight weeks old) and female control mouse (PRCre/+, n=9/group, eight weeks old). At estrus cycle in 4th weeks of endometriosis induction, we harvested ectopic lesions and then isloated total RNA from them. After that, we combine total RNA from three eutopic endometria to form a single group. Therefore, we made three RNA groups from nine ERβ overexpressing eutopic endometrium and nine eutopic control endometrium. Transcriptome analysis in the uterus: We harvested uteri ERBOE female mouse, and it's female control mouse (PRCre/+) (n=9/group, 12 weeks old) at estrus cycle and isolated total RNA from them. We combined total RNA from three uteri to form a single group. Therefore, we made three RNA groups from nine ERβ overexpressing uteri and nine control uteri.