Expression of MUC1 mucin gene by hamster tracheal surface epithelial cells in primary culture.
Expression of MUC1 mucin gene by hamster tracheal surface epithelial cells in primary culture.
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原代培养的仓鼠气管表面上皮细胞MUC1粘蛋白基因的表达。
DOI：
10.1165/ajrcmb.15.2.8703480
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Kim,KC
中科院分区：
文献类型：
--
作者：
Park,H;Hyun,SW;Kim,KC
原代仓鼠气管表面上皮细胞（HTSE）在根尖表面携带黏液样糖蛋白，可由中性粒细胞弹性蛋白酶释放。在一些癌细胞中，粘蛋白定位于细胞表面，并被证明是由MUC1粘蛋白基因编码的。本实验的目的是：(1)确定HTSE细胞是否表达MUC1粘蛋白基因；(2)分离鼠MUC1互补DNA （cDNA）并对其进行鉴定；(3)检测MUC1 mRNA在不同培养阶段的表达模式。基于人和小鼠MUC1基因的同源序列，利用退化引物逆转录聚合酶链反应扩增HTSE细胞rna，发现存在一个与小鼠MUC1基因序列相似度为88%的cDNA （0.5 kb）。以该0.5 kb cDNA为探针，筛选HTSE细胞cDNA文库，分离出仓鼠MUC1 cDNA克隆。序列分析表明，该cDNA编码一个676个氨基酸的完整膜蛋白，由(1)n端信号序列，(2)串联重复序列，编码20个氨基酸的12个重复序列，以及(3)c端由退化串联重复序列和一个包含跨膜和细胞质结构域的独特序列组成。细胞质结构域中存在7个酪氨酸残基，表明其可能是一种受体。最后，HTSE细胞中MUC1粘蛋白基因的表达似乎与分泌细胞的分化有关。
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.