ESTROGEN REGULATES THE SYNTHESIS OF EPIDERMAL GROWTH-FACTOR IN MOUSE UTERINE EPITHELIAL-CELLS
ESTROGEN REGULATES THE SYNTHESIS OF EPIDERMAL GROWTH-FACTOR IN MOUSE UTERINE EPITHELIAL-CELLS
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DOI：
10.1210/mend-4-3-510
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发表时间：
1990-03-01
影响因子：
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通讯作者：
DEY, SK
中科院分区：
文献类型：
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作者：
HUETHUDSON, YM;CHAKRABORTY, C;DEY, SK
免疫细胞化学分析，使用几种小鼠表皮生长因子（EGF）的多克隆抗体，检测到的免疫反应性仅在子宫腔和腺上皮上晚发情前期，发情期，早期妊娠第1天，但不晚于第1天。在卵巢切除子宫中未检测到这种免疫反应性，但在雌激素刺激后，在12-24 h之间首先在腔上皮中检测到，然后在48 h时在腺上皮中也检测到。雌激素停药72小时后，EGF免疫反应性不再检测到。这种反应对雌激素是特异性的，在注射孕酮（2 mg/天，持续4天）后没有发生。使用preproEGF特异性的抗肽抗体，在卵巢切除的子宫中未检测到免疫反应性，在雌激素化的子宫和颌下腺中检测到弱反应性，在肾脏中检测到强反应性。子宫RNA的北方印迹分析未能检测到预期的4.8-磷酸酶prepro-EGF mRNA，而是检测到罕见的2.4磷酸酶转录物，这表明EGF mRNA在子宫中交替加工。EGF编码的子宫转录本的存在下，进一步证明了EGF编码区特异性寡脱氧核糖核苷酸（oligo）的聚合酶链反应扩增的子宫cDNA的杂交。原位杂交，使用prepro-EGF cRNA探针以及EGF编码区特异性寡核苷酸，显示与EGF免疫染色（上皮细胞）共定位的杂交，并且不存在于非EGF免疫反应性细胞中。脉冲标记实验加上免疫亲和层析表明，雌激素诱导的酸溶性免疫反应性蛋白，这是相同的大小为正宗的EGF的合成的相对速率增加。此外，通过DEAE-纤维素色谱分离的酸溶性子宫蛋白的分析表明，抗原活性和受体结合活性的单一重合峰，其与正宗的EGF从柱中共洗脱。电子显微镜定位EGF免疫反应性的腔上皮细胞的高尔基体。总之，这些结果表明，雌激素调节EGF基因的表达，特别是在子宫上皮细胞。该基因的表达增加导致该蛋白质合成的相对速率增加和成熟EGF的积累。
Immunocytochemical analyses, using several mouse epidermal growth factor (EGF) polyclonal antibodies, detected immunoreactivity only in uterine luminal and glandular epithelia on late proestrus, estrus, and early on day 1 of pregnancy, but not late on day 1. This immunoreactivity was not detected in the ovariectomized uterus, but after estrogen stimulation it was detected first in the luminal epithelium between 12-24 h and then also in the glandular epithelium by 48 h. After 72 h of estrogen withdrawal, EGF immunoreactivity was no longer detected. This response was specific for estrogen and did not occur after progesterone injection (2 mg/day for 4 days). Using antipeptide antibodies specific for preproEGF, no immunoreactivity was detected in the ovariectomized uterus, weak reactivity was detected in the estrogenized uterus and submandibular gland and strong reactivity was detected in the kidney. Northern blot analysis of uterine RNA failed to detect the expected 4.8-kilobase prepro-EGF mRNA, but, instead, a rare transcript of 2.4 kilobases was detected, which suggests that EGF mRNA is alternately processed in the uterus. The presence of an EGF-coding uterine transcript was further documented by hybridization of an EGF-coding region-specific oligodeoxyribonucleotide (oligo) to polymerase chain reaction-amplified uterine cDNA. In situ hybridization, using a prepro-EGF cRNA probe as well as an EGF-coding region-specific oligo, showed hybridization that colocalized with the EGF immunostaining (epithelia) and was absent from non-EGF-immunoreactive cells. Pulse labelling experiments coupled with immunoaffinity chromatography showed that estrogen induced an increase in the relative rate of synthesis of an acid-soluble immunoreactive protein which was the same size as authentic EGF. Furthermore, analysis of acid-soluble uterine proteins fractionated by DEAE-cellulose chromatography demonstrated a single coincident peak of antigenic activity and receptor-binding activity which coeluted from the column with authentic EGF. Electron microscopy localized EGF immunoreactivity to the Golgi of luminal epithelial cells. Taken together these results suggest that estrogen regulates expression of the EGF gene specifically in uterine epithelial cells. Increased expression of this gene results in an increase in the relative rate of synthesis of this protein and the accumulation of mature EGF.