Cloning and characterization of the cDNA and gene for human epitheliasin
Cloning and characterization of the cDNA and gene for human epitheliasin
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DOI：
10.1046/j.1432-1327.2001.02165.x
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发表时间：
2001-05-01
期刊：
影响因子：
--
通讯作者：
Hoidal, JR
中科院分区：
文献类型：
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作者：
Jacquinet, E;Rao, NV;Hoidal, JR
在此之前，我们报告了小鼠基因，上皮素的克隆和表征。在本工作中，我们克隆了全长人类直向同源物的cDNA，并对其基因进行了表征，包括2kb的5'侧翼序列。对成人组织中上皮素基因表达的分析表明，它表达为3.4kb和2kb的转录本。主要的3.4kb转录物按以下顺序观察：前列腺>结肠>小肠>胰腺>肾>肺>肝。胎儿组织中的Epitheliasin转录本仅在肾和肺中观察到。组织原位杂交分析显示，上皮素优先表达于上皮细胞。该基因由14个外显子和13个内含子组成。在5'侧翼区，我们鉴定了两个转录起始位点和三个CpG岛，包括许多潜在的调控元件，包括SP1，SREBP，GRE/PRE和ERE。转录位点的上游区域缺少TATA盒，但含有启动子样元件以及下游启动子样元件。前列腺淋巴结癌（LNCaP）细胞的体外实验表明，上皮细胞素基因诱导雄激素和诱导不被放线菌酮，表明诱导不需要中间蛋白质因子阻断。免疫沉淀分析表明，雄激素强烈增加epitheliasin蛋白水平。
Previously, we reported cloning and characterization of the mouse gene, epitheliasin. In the present work we cloned the cDNA of the full-length human orthologue and characterized its gene including 2 kb of 5' flanking sequence. Analysis of epitheliasin gene expression in adult tissues shows that it is expressed as 3.4 kb and 2 kb transcripts. The major 3.4 kb transcript is observed in the following order: prostate > colon > small intestine > pancreas > kidney > lung > liver. Epitheliasin transcripts in fetal tissues are observed only in kidney and lung. In situ hybridization analysis of tissues revealed that epitheliasin was preferentially expressed in epithelial cells. The gene consists of 14 exons and 13 introns based on comparison with its cDNA sequence. In the 5' flanking region, we identified two transcription start sites and three CpG islands encompassing a number of potential regulatory elements including SP1, SREBP, GRE/PRE and ERE. The region upstream of the transcription sites lacks a TATA box but contains an initiator-like element as well as a downstream promoter-like element. In vitro experiments with lymph node carcinoma of prostate (LNCaP) cells revealed that the epitheliasin gene was induced by androgens and the induction was not blocked by cycloheximide indicating that the induction required no intermediate protein factors. Immunoprecipitation analysis showed that androgens strongly increased epitheliasin protein levels.