Ace C I, Okulicz W C
Department of Obstetrics and Gynecology, University of Massachusetts Medical School, Worcester 01655, USA.
Mol Cell Endocrinol. 1995 Nov 30;115(1):95-103. doi: 10.1016/0303-7207(95)03674-v.
During the shift from a proliferative to a secretory endometrium in the rhesus menstrual cycle, progesterone action causes massive metabolic and structural remodelling. In order to identify genes whose expression is potentially important for the change from estrogen (E) to progesterone (P) dominance we have initiated a study of specific gene regulation using semiquantitative, reverse transcription polymerase chain reaction (RT-PCR). PolyA+ RNA was isolated from both E-dominant (days 9-13 of artificial menstrual cycles [AMCs]) and P-dominant (days 21-23) rhesus monkey endometria. The two pools of mRNA were converted to cDNA, end-ligated to double-stranded oligonucleotide adaptors and amplified by PCR using an adaptor-complementary primer. This procedure resulted in the production of E- and PcDNA template populations for cDNA-specific screening and comparative quantitation by PCR. Initial analysis showed that placental protein 14 (PP14) was P-dependent and human complement 3 (HC3) was up-regulated in E-dominant tissue, whereas the housekeeping genes B-actin and glyceraldehyde-3-phosphate dehydrogenase (G-3-PDH) were expressed at equivalent levels under E and P dominance. Expression of the E receptor (ER), P receptor (PR), epidermal growth factor receptor (EGFR) and insulin-like growth factor (IGF-I) was equivalent under E or P dominance. Expression of epidermal growth factor (EGF) and retinoblastoma (RB) was down-regulated in P-dominant tissue. Conversely IGF-1 receptor (IGF-1-R), transforming growth factor-beta 2 (TGFB-2), TGFB-2 receptor (TGFB-2-R), 17 beta-hydroxysteroid dehydrogenase (17-B-HSD) and leukemia inhibitory factor (LIF) levels were up-regulated in PcDNA. Among these factors, PP14, LIF, IGF-1-R TGFB-2 and 17-B-HSD were also detectable in PCR in a P-dependent cDNA library isolated by subtractive hybridization. These data provide evidence for hormonal regulation of specific gene products that may play important roles in the normal maturation of the primate endometrium in preparation for implantation.
在恒河猴月经周期中，子宫内膜从增殖期向分泌期转变的过程中，孕酮作用会引发大规模的代谢和结构重塑。为了确定那些其表达可能对从雌激素（E）主导转变为孕酮（P）主导至关重要的基因，我们启动了一项使用半定量逆转录聚合酶链反应（RT-PCR）研究特定基因调控的实验。从处于E主导阶段（人工月经周期[AMCs]的第9 - 13天）和P主导阶段（第21 - 23天）的恒河猴子宫内膜中分离出PolyA + RNA。这两组mRNA被转化为cDNA，末端连接双链寡核苷酸接头，并使用接头互补引物通过PCR进行扩增。此过程产生了用于cDNA特异性筛选和通过PCR进行比较定量的E和P cDNA模板群体。初步分析表明，胎盘蛋白14（PP14）依赖于P，人补体3（HC3）在E主导的组织中上调，而管家基因β-肌动蛋白和甘油醛-3-磷酸脱氢酶（G-3-PDH）在E和P主导下表达水平相当。雌激素受体（ER）、孕激素受体（PR）、表皮生长因子受体（EGFR）和胰岛素样生长因子（IGF-I）在E或P主导下的表达相当。表皮生长因子（EGF）和成视网膜细胞瘤（RB）在P主导的组织中表达下调。相反，IGF-1受体（IGF-1-R）、转化生长因子-β2（TGFB-2）、TGFB-2受体（TGFB-2-R）、17β-羟基类固醇脱氢酶（17-B-HSD）和白血病抑制因子（LIF）水平在P cDNA中上调。在这些因子中，PP14、LIF、IGF-1-R、TGFB-2和17-B-HSD在通过消减杂交分离的P依赖cDNA文库的PCR中也可检测到。这些数据为特定基因产物的激素调控提供了证据，这些基因产物可能在灵长类子宫内膜为着床做准备的正常成熟过程中发挥重要作用。
