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人绒毛膜癌细胞系中内源性胎盘生长激素变体和绒毛膜生长催乳素基因的组织特异性表达及甲状腺激素调节

Tissue-specific expression and thyroid hormone regulation of the endogenous placental growth hormone variant and chorionic somatomammotropin genes in a human choriocarcinoma cell line.

作者信息

Nickel B E, Cattini P A

机构信息

Department of Physiology, University of Manitoba, Winnipeg, Canada.

出版信息

Endocrinology. 1991 May;128(5):2353-9. doi: 10.1210/endo-128-5-2353.

Abstract

Human (h) placenta-derived choriocarcinoma cell lines (BeWo, JAR, and JEG-3) were examined for expression of pituitary GH (hGH-N) as well as placental GH variant (hGH-V) and chorionic somatomammotropin (hCS, encoded by the hCS-A or hCS-B gene). RNA was isolated and assessed using hGH-N complementary DNA since hGH and hCS genes share more than 90% sequences similarity. The relative expression is BeWo greater than JAR greater than JEG-3. In BeWo cells expression of placental hCS-A, hCS-B, and hGH-V genes, but not pituitary hGH-N, is observed using polyadenylated RNA and oligonucleotide probes specific for the different family members. The absence of hGH-N expression in BeWo cells is not due to deletion or gross rearrangement of the gene. No difference was seen between the hGH/hCS genes in genomic DNA from these cells and the DNA from placenta and pituitary when analyzed by restriction digestion and blotting. Treatment of BeWo cells with 10 nM T3 results in a 6-fold increase in messenger RNA from placental members of the hGH gene family. Levels of hCS-A, hCS-B, and hGH-V transcripts are all elevated. Cellular and secreted proteins from BeWo cells were analyzed by Western blotting, and a band of about 22 kilodaltons was detected using a polyclonal antibody which cross-reacts with hGH-V and hCS. The level of 22 kilodalton band in samples of cellular as well as released protein was increased by T3 treatment. BeWo cells provide a model system for studying hGH-V and hCS regulation as well as tissue-specific expression.

摘要

对源自人胎盘的绒毛膜癌细胞系(BeWo、JAR和JEG-3)进行了检测,以确定垂体生长激素(hGH-N)、胎盘生长激素变体(hGH-V)和绒毛膜促生长催乳素(hCS,由hCS-A或hCS-B基因编码)的表达情况。由于hGH和hCS基因的序列相似度超过90%,因此使用hGH-N互补DNA分离并评估RNA。相对表达水平为BeWo大于JAR大于JEG-3。在BeWo细胞中,使用聚腺苷酸化RNA和针对不同家族成员的寡核苷酸探针,可观察到胎盘hCS-A、hCS-B和hGH-V基因的表达,但未观察到垂体hGH-N的表达。BeWo细胞中hGH-N表达的缺失并非由于该基因的缺失或大规模重排。通过限制性消化和印迹分析,这些细胞的基因组DNA中的hGH/hCS基因与胎盘和垂体的DNA之间未发现差异。用10 nM T3处理BeWo细胞会导致hGH基因家族胎盘成员的信使RNA增加6倍。hCS-A、hCS-B和hGH-V转录本的水平均升高。通过蛋白质印迹法分析了BeWo细胞的细胞蛋白和分泌蛋白,使用与hGH-V和hCS发生交叉反应的多克隆抗体检测到一条约22千道尔顿的条带。T3处理使细胞蛋白和释放蛋白样品中22千道尔顿条带的水平升高。BeWo细胞为研究hGH-V和hCS的调控以及组织特异性表达提供了一个模型系统。

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