Can G, Abdel-Malek Z, Porter-Gill P A, Gill P, Boyce S, Grabowski G A, Nordlund J, Farooqui J
Department of Dermatology, University of Cincinnati, Ohio, USA.
J Invest Dermatol. 1998 Sep;111(3):485-91. doi: 10.1046/j.1523-1747.1998.00315.x.
Proopiomelanocortin (POMC) is a precursor polypeptide for various bioactive peptides, including adrenocorticotropic hormone, alpha-, beta-, and gamma-melanotropin, beta-endorphin, and beta-lipotropin. Although the classical source of POMC is the pituitary, various studies indicate the expression of POMC in several nonpituitary tissues. In this study, in situ hybridization with anti-sense cRNA riboprobe was used to show expression of POMC mRNA in human epidermis and cultured human epidermal cells (melanocytes and keratinocytes). POMC mRNA was amplified by reverse transcriptase-polymerase chain reaction using anti-sense and sense primers designed from Exons 2 and 3 of POMC gene. A approximately 300 bp product was present in normal human skin, grafted human skin, and cultured normal human melanocytes and keratinocytes. By Southern analysis this product was hybridized specifically to the POMC cDNA. Sequence analysis of the reverse transcriptase polymerase chain reaction product from tissues or cells showed 85% homology to POMC cDNA from human, bovine, pig, and monkey sources. This suggests the existence of a putative isoform or variant of POMC mRNA in human epidermis.
阿黑皮素原(POMC)是多种生物活性肽的前体多肽,包括促肾上腺皮质激素、α-、β-和γ-促黑素、β-内啡肽和β-促脂素。虽然POMC的经典来源是垂体,但各种研究表明POMC在几种非垂体组织中也有表达。在本研究中,使用反义cRNA核糖探针进行原位杂交,以显示POMC mRNA在人表皮和培养的人表皮细胞(黑素细胞和角质形成细胞)中的表达。使用从POMC基因外显子2和3设计的反义引物和正义引物,通过逆转录聚合酶链反应扩增POMC mRNA。在正常人皮肤、移植的人皮肤以及培养的正常人黑素细胞和角质形成细胞中存在一个约300 bp的产物。通过Southern分析,该产物与POMC cDNA特异性杂交。对来自组织或细胞的逆转录聚合酶链反应产物进行序列分析,结果显示与来自人、牛、猪和猴的POMC cDNA有85%的同源性。这表明人表皮中存在一种假定的POMC mRNA同工型或变体。