Sanno N, Jin L, Qian X, Osamura R Y, Scheithauer B W, Kovacs K, Lloyd R V
Department of Laboratory Medicine and Pathology, Mayo Clinic and Mayo Foundation, Rochester, Minnesota 55905, USA.
J Clin Endocrinol Metab. 1997 Jun;82(6):1974-82. doi: 10.1210/jcem.82.6.3976.
In the nontumorous pituitary, GnRH stimulates the release and synthesis of LH and FSH by gonadotroph cells via the GnRH receptor (GnRH-R). Little is known, however, about expression of GnRH and GnRH-R messenger RNAs (mRNAs) in nontumorous pituitary tissue and in adenomas. To learn more about the distribution and regulatory roles of GnRH and its receptor, we investigated the expression of both GnRH and GnRH-R mRNAs in nontumorous human pituitary and in various types of pituitary adenomas using the RT-PCR, in situ hybridization, and in situ hybridization in combination with RT-PCR (in situ RT-PCR). Using RT-PCR, GnRH mRNA was found to be expressed in normal human pituitaries and in all types of adenomas. Similarly, GnRH-R mRNA was expressed in nontumorous human pituitaries and in most, but not all, adenomas. These included 5 gonadotroph adenomas, 6 null cell adenomas, 1 of 2 GH-producing tumors, and 1 of 2 ACTH-producing adenomas, but not in the 2 PRL-producing adenomas examined. In situ hybridization studies showed GnRH and GnRH-R mRNAs in all 3 nontumorous pituitaries and in 12 of 33 (36.4%) and 10 of 33 adenomas (30.3%), respectively. Using an indirect in situ RT-PCR technique to increase the sensitivity of the in situ localization, GnRH and GnRH-R mRNAs were detected in 29 (87.9%) and 25 (75.8%) of 33 adenomas, respectively. This is the first report of the localization of GnRH and GnRH-R mRNAs in individual pituitary adenoma cells using in situ RT-PCR. The frequent expression of GnRH and GnRH-R mRNAs in pituitary cells suggests that GnRH has autocrine/paracrine functions in nontumorous and neoplastic pituitary tissues.
在非肿瘤性垂体中,促性腺激素释放激素(GnRH)通过GnRH受体(GnRH-R)刺激促性腺激素细胞释放和合成促黄体生成素(LH)和促卵泡生成素(FSH)。然而,关于GnRH及其信使核糖核酸(mRNA)在非肿瘤性垂体组织和腺瘤中的表达情况,人们了解甚少。为了进一步了解GnRH及其受体的分布和调节作用,我们运用逆转录聚合酶链反应(RT-PCR)、原位杂交以及原位杂交结合RT-PCR(原位RT-PCR)技术,研究了GnRH和GnRH-R mRNA在非肿瘤性人类垂体及各类垂体腺瘤中的表达。运用RT-PCR技术发现,GnRH mRNA在正常人类垂体及所有类型的腺瘤中均有表达。同样,GnRH-R mRNA在非肿瘤性人类垂体及大多数(但并非全部)腺瘤中也有表达。这些腺瘤包括5例促性腺激素细胞腺瘤、6例无功能细胞腺瘤、2例生长激素分泌性肿瘤中的1例以及2例促肾上腺皮质激素分泌性腺瘤中的l例,但在所检测的2例催乳素分泌性腺瘤中未发现表达。原位杂交研究显示,在所有3例非肿瘤性垂体以及33例腺瘤中的12例(36.4%)和10例(30.3%)中分别检测到了GnRH和GnRH-R mRNA。运用间接原位RT-PCR技术提高原位定位的敏感性,在33例腺瘤中的29例(87.9%)和25例(75.8%)中分别检测到了GnRH和GnRH-R mRNA。这是首次运用原位RT-PCR技术对单个垂体腺瘤细胞中GnRH和GnRH-R mRNA进行定位的报道。GnRH和GnRH-R mRNA在垂体细胞中的频繁表达表明,GnRH在非肿瘤性和肿瘤性垂体组织中具有自分泌/旁分泌功能。