Kakar S S, Jennes L
Department of Physiology and Biophysics, University of Alabama at Birmingham 35294, USA.
Cancer Lett. 1995 Nov 27;98(1):57-62.
Recently, cloning of the gonadotropin-releasing hormone (GnRH) receptor from the human breast tumor cell line (MCF-7) and from an ovarian tumor, and its expression in various other human tumors, tumor cell lines and reproductive organs have been reported (Kakar et al., Mol. Cell. Endocrinol., 106 (1994) 145-149). In the present studies, we investigated the expression of GnRH and GnRH receptor mRNAs in normal human non-reproductive tissues. Using reverse transcriptase-polymerase chain reaction (RT-PCR) techniques and specific oligonucleotide primers derived from the placental GnRH cDNA sequence, PCR products of the expected size were obtained from human liver, heart, skeletal muscle, kidney, placenta, and pituitary. The authenticity of the PCR products was confirmed by Southern blot analysis with an internal oligonucleotide primer as probe. Similarly, using specific oligonucleotide primers for the GnRH receptor selected from the human pituitary GnRH receptor cDNA sequence, PCR products of the expected size were amplified from human liver, heart, skeletal muscle, kidney, placenta, and pituitary, and these strongly hybridized with the human GnRH receptor cDNA on Southern blot. Cloning and nucleotide sequencing of the PCR products for the GnRH and GnRH receptor from heart revealed identical sequences when compared to the human placental GnRH and pituitary GnRH receptor cDNAs, respectively. These data demonstrate for the first time the existence of GnRH and GnRH receptor mRNAs in normal human non-reproductive tissues and suggest that GnRH and its receptor may play an important role in the regulation of cellular functions in an autocrine or paracrine manner, in addition to regulating the secretion of gonadotropins from the anterior pituitary.
最近，已报道了从人乳腺肿瘤细胞系（MCF - 7）和一种卵巢肿瘤中克隆促性腺激素释放激素（GnRH）受体，以及其在各种其他人类肿瘤、肿瘤细胞系和生殖器官中的表达（卡卡等人，《分子与细胞内分泌学》，106卷（1994年）145 - 149页）。在本研究中，我们调查了GnRH和GnRH受体mRNA在正常人类非生殖组织中的表达。使用逆转录聚合酶链反应（RT - PCR）技术以及源自胎盘GnRH cDNA序列的特异性寡核苷酸引物，从人肝脏、心脏、骨骼肌、肾脏、胎盘和垂体中获得了预期大小的PCR产物。用内部寡核苷酸引物作为探针通过Southern印迹分析证实了PCR产物的真实性。同样，使用从人垂体GnRH受体cDNA序列中选择的GnRH受体特异性寡核苷酸引物，从人肝脏、心脏、骨骼肌、肾脏、胎盘和垂体中扩增出预期大小的PCR产物，并且这些产物在Southern印迹上与人类GnRH受体cDNA强烈杂交。来自心脏的GnRH和GnRH受体的PCR产物的克隆和核苷酸测序显示，与人类胎盘GnRH和垂体GnRH受体cDNA相比，分别具有相同的序列。这些数据首次证明了正常人类非生殖组织中存在GnRH和GnRH受体mRNA，并表明GnRH及其受体除了调节垂体前叶促性腺激素的分泌外，可能以自分泌或旁分泌方式在细胞功能调节中发挥重要作用。
