Park H, Hyun S W, Kim K C
Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore 21201, USA.
Am J Respir Cell Mol Biol. 1996 Aug;15(2):237-44. doi: 10.1165/ajrcmb.15.2.8703480.
Primary hamster tracheal surface epithelial (HTSE) cells carry mucin-like glycoproteins on the apical surface which are releasable by neutrophil elastase. In some cancer cells, mucins are localized on the cell surface and have been shown to be encoded by the MUC1 mucin gene. The objectives of the present experiments were: (I) to determine if HTSE cells express MUC1 mucin gene; (2) if they do, to isolate and characterize the hamster MUC1 complementary DNA (cDNA); and (3) to examine the pattern of MUC1 mRNA expression at different stages of culture. Reverse transcriptase-polymerase chain reaction amplification of HTSE cell RNAs using degenerate primers based on homologous sequences between the human and mouse MUC1 genes revealed the presence of a cDNA (0.5 kb) which has an 88% similarity in sequence with the mouse MUC1 cDNA. Using this 0.5 kb cDNA as a probe, an HTSE cell cDNA library was screened to isolate a hamster MUC1 cDNA clone. Sequence analysis of the cDNA revealed that it encodes an integral membrane protein of 676 amino acids which consists of (1) an N-terminal signal sequence, (2) the tandem repeat domain encoding 12 repeats of 20 amino acids, and (3) the C-terminal region consisting of degenerate tandem repeats and a unique sequence containing both the transmembrane and cytoplasmic domains. The presence of seven tyrosine residues in the cytoplasmic domain suggests a potential role as a receptor. Finally, expression of MUC1 mucin gene in HTSE cells appears to be associated with differentiation of secretory cells.
原代仓鼠气管表面上皮(HTSE)细胞在顶端表面携带黏蛋白样糖蛋白,这些糖蛋白可被中性粒细胞弹性蛋白酶释放。在一些癌细胞中,黏蛋白定位于细胞表面,并且已证明由MUC1黏蛋白基因编码。本实验的目的是:(1)确定HTSE细胞是否表达MUC1黏蛋白基因;(2)如果表达,分离并鉴定仓鼠MUC1互补DNA(cDNA);(3)检查培养不同阶段MUC1 mRNA的表达模式。使用基于人和小鼠MUC1基因同源序列的简并引物对HTSE细胞RNA进行逆转录酶 - 聚合酶链反应扩增,发现存在一个cDNA(0.5 kb),其序列与小鼠MUC1 cDNA有88%的相似性。使用这个0.5 kb cDNA作为探针,筛选HTSE细胞cDNA文库以分离仓鼠MUC1 cDNA克隆。对该cDNA的序列分析表明,它编码一个由676个氨基酸组成的整合膜蛋白,该蛋白由(1)一个N端信号序列,(2)编码20个氨基酸的12个重复序列的串联重复结构域,以及(3)由简并串联重复序列和包含跨膜和细胞质结构域的独特序列组成的C端区域组成。细胞质结构域中七个酪氨酸残基的存在表明其可能作为受体发挥作用。最后,HTSE细胞中MUC1黏蛋白基因的表达似乎与分泌细胞的分化有关。