Overexpression of estrogen receptor-alpha gene suppresses gap junctional intercellular communication in endometrial carcinoma cells.
Stimulation of the endometrium by estrogens without the differentiating effect of progestins is the primary etiological factor associated with the development of endometrial hyperplasia and adenocarcinoma. However, the correlation between sex steroids and gap junctional intercellular communication (GJIC), which is considered to play an important role in the control of cell growth and differentiation, is not well known in endometrial carcinoma. In this study, we focused on the influence of estrogen and its receptor in connexin (Cx) expression and GJIC in endometrial carcinoma cells, established stable clone IK-ER1 overexpressing ER-alpha to transfect the expression vector and analysed them in various hormonal conditions. The growth of IK-ER1 was accelerated by 17beta-estradiol and the acceleration of the 5-bromo-25-deoxyuridine labeling index was observed. GJIC was assayed by scoring the number of dye-coupled cells after microinjection of single cells with Lucifer-Yellow, and subcellular localization of Cx26 and Cx32 was analysed by immunocytochemistry. In the presence of estradiol, dye-coupled cells of IK-ER1 were significantly reduced compared to those without estradiol and the reduction was completely inhibited by adding ICI182.780, a pure antiestrogen substrate. Cxs were detected as only small spots by immunocytochemistry, and Western blotting showed that the expression was decreased. These results suggest that activation of ER-alpha by estrogen results in tumor progression by stimulating cell growth and suppressing GJIC via suppression of the expression of Cxs in endometrial carcinogenesis.
Saito
Tsuyoshi
T
Department of Obstetrics and Gynecology, Sapporo Medical University, School of Medicine, S-1, W-16, Chuo-ku, Sapporo 060-8543, Japan. tsaito@sapmed.ac.jp
Tanaka
Ryoichi
R
Wataba
Koya
K
Kudo
Ryuichi
R
Yamasaki
Hiroshi
H
D002450:Cell Communication; D002455:Cell Division / Q000187:drug effects; D045744:Cell Line, Tumor; D000072259:Connexin 26; D017630:Connexins / Q000032:analysis / Q000378:metabolism; D016889:Endometrial Neoplasms / Q000378:metabolism* / Q000473:pathology; D004958:Estradiol / Q000031:analogs & derivatives* / Q000494:pharmacology; D004965:Estrogen Antagonists / Q000494:pharmacology; D047628:Estrogen Receptor alpha; D005260:Female; D005456:Fluorescent Dyes; D000077267:Fulvestrant; D017629:Gap Junctions / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007546:Isoquinolines; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism*; D000097002:Gap Junction beta-1 Protein
D000072259:Connexin 26; D017630:Connexins; D004958:Estradiol; D004965:Estrogen Antagonists; D047628:Estrogen Receptor alpha; D005456:Fluorescent Dyes; D000077267:Fulvestrant; D007546:Isoquinolines; D011960:Receptors, Estrogen; D000097002:Gap Junction beta-1 Protein; C000607042:GJB2 protein, human; C017475:lucifer yellow