Chen Fang-Ping, Hsu Todd, Hu Chin-Hwa, Wang Wen-Der, Wang Kun-Chuang, Teng Li-Fen
Department of Obstetrics and Gynecology, Chang Gung Memorial Hospital, Keelung, Taiwan 204, ROC.
Chang Gung Med J. 2004 Feb;27(2):107-15.
Postmenopausal osteoporosis is associated with estrogen deficiency. Estrogens have effects on bone metabolism, which are mediated by estrogen receptors (ERs). If estrogen responsiveness is related to the ER expression level, ER expression in postmenopausal women should be different from previous studies using osteoblast lineage. We investigated the expression of variant isoforms of ER messenger ribonucleic acid (mRNA) in osteoblasts (OB) from postmenopausal women and a human osteosarcoma cell line, MG 63.
Osteoblast cultures were prepared from the upper femur of postmenopausal patients or MG 63. For OB cultures at 5, 10, 15, 20, and 25 days, the expressions of ERalpha and beta mRNA were examined using reverse transcriptase-polymerase chain reaction.
In MG 63, ERbeta mRNA was constantly and highly expressed during the 25-day culture, whereas ERalpha mRNA was barely detected. In the primary OB cells, both ERalpha and beta mRNA were transcribed during the 25-day culture, but expression of ERalpha mRNA was much stronger than that of ERbeta mRNA. A splice variant form of ERbeta mRNA that was missing the entire exon 2 (ERbeta delta 2) was detected and heterogeneously expressed in OB cultures from 16 postmenopausal women.
Differential expressions of these ER isoforms suggest that they may have different functions or that they interact with each other during bone metabolism. The different ratio of ERbeta to ERbeta delta 2 mRNA or ERalpha to ERbeta mRNA expressions in osteoblast cultures may be related to different bone conditions. Whether the presence of ERbeta delta 2 in postmenopausal women influences the biological properties of bone needs to be determined.
绝经后骨质疏松症与雌激素缺乏有关。雌激素对骨代谢有影响,这种影响由雌激素受体(ERs)介导。如果雌激素反应性与ER表达水平相关,那么绝经后女性的ER表达应该与之前使用成骨细胞系的研究有所不同。我们研究了绝经后女性成骨细胞(OB)和人骨肉瘤细胞系MG 63中雌激素受体信使核糖核酸(mRNA)变异体亚型的表达。
从绝经后患者的股骨上段或MG 63制备成骨细胞培养物。对于培养5、10、15、20和25天的OB培养物,使用逆转录聚合酶链反应检测ERα和β mRNA的表达。
在MG 63中,ERβ mRNA在25天的培养过程中持续高表达,而ERα mRNA几乎未被检测到。在原代OB细胞中,ERα和β mRNA在25天的培养过程中均有转录,但ERα mRNA的表达比ERβ mRNA强得多。检测到一种缺失整个外显子2的ERβ mRNA剪接变异体形式(ERβδ2),并在16名绝经后女性的OB培养物中异质性表达。
这些ER亚型的差异表达表明它们可能具有不同的功能,或者在骨代谢过程中相互作用。成骨细胞培养物中ERβ与ERβδ2 mRNA或ERα与ERβ mRNA表达的不同比例可能与不同的骨状况有关。绝经后女性中ERβδ2的存在是否会影响骨的生物学特性有待确定。