CAJ | 학술논문

目的：大鼠 UREB1基因编码的蛋白质能特异性地结合位于强吗啡基因启动子上游的一段 DNA序列 (upstream regulatory element,URE)。早期研究证实 UREB1对强吗啡基因启动子的转录具有促进作用，而对 p53的转录激活作用具有抑制效应。本实验目的就是克隆人 UREB1基因，探索其与肿瘤发生发展的关系。方法：利用人工合成寡核苷酸探针从人脑 cDNA文库中筛选人 UREB1基因，应用大肠杆菌表达的重组蛋白质制备的抗体检测肿瘤组织中 UREB1的表达分布。结果：获得了人 UREB1基因，核苷酸序列在对应区域及氨基酸序列与大鼠 UREB1基因 cDNA序列和氨基酸序列皆有 91％的同源性。在各种肿瘤组织中都有 UREB1的表达，但是表达水平及定位不一样。初步发现有这样一个规律，随着肿瘤的恶性程度增加 UREB1在核内的聚集程度增加。 UREB1的酪氨酸磷酸化分析结果显示，肿瘤恶性程度高， UREB1的酪氨酸磷酸化程度高。结论： UREB1可能参与肿瘤的发生发展，其酪氨酸磷酸化水平可以影响肿瘤的恶性程度。
목적：대서 UREB1기인편마적단백질능특이성지결합위우강마배기인계동자상유적일단 DNA서렬 (upstream regulatory element,URE)。조기연구증실 UREB1대강마배기인계동자적전록구유촉진작용，이대 p53적전록격활작용구유억제효응。본실험목적취시극륭인 UREB1기인，탐색기여종류발생발전적관계。방법：이용인공합성과핵감산탐침종인뇌 cDNA문고중사선인 UREB1기인，응용대장간균표체적중조단백질제비적항체검측종류조직중 UREB1적표체분포。결과：획득료인 UREB1기인，핵감산서렬재대응구역급안기산서렬여대서 UREB1기인 cDNA서렬화안기산서렬개유 91％적동원성。재각충종류조직중도유 UREB1적표체，단시표체수평급정위불일양。초보발현유저양일개규률，수착종류적악성정도증가 UREB1재핵내적취집정도증가。 UREB1적락안산린산화분석결과현시，종류악성정도고， UREB1적락안산린산화정도고。결론： UREB1가능삼여종류적발생발전，기락안산린산화수평가이영향종류적악성정도。
Objective：Rat UREB1 protein coded by the gene UREB1 can specially bind to URE (upstream regulatory element) which is in the upstream of the promoter. It′ s reported that the protein of UREB1 promote the transcription of Dynorphin gene and inhibits p53 transactivation. This study was designed to clone human UREB1 gene and explore the relationship between UREB1 and the development of tumor. Methods： The artificial synthetic oligonucleotide was used as the probe to screen human brain cDNA library and human UREB1 gene was cloned. The antibody, which was produced using the recombinant UREB1 from E.coli as the antigen and immunizing the animals, was utilized for detecting the distribution of UREB1 in different tumor tissues. Results： The human UREB1 gene was cloned by using in situ hybridization for screening human brain cDNA library, and the nucleotide sequences and the deduced amino acid sequence of human UREB1 has 91％ homology with that of rat UREB1 identified previously. Western blot analysis revealed that the human UREB1 was present in all tumor tissues but the quantity of UREB1 in different tissues was not the same. Immunohistochemistry results shown that the human UREB1 distributes primarily in the cytoplasm and nuclear of tumor cells and nuclear UREB1 in carcinosarcoma is much more than that in adenoma. After analyzing the level of tyrosine phosphorylated UREB1 in a few tumor tissues, the result shown that the more malignant the tumor tissue was, the higher level the tyrosine phosphorylated of UREB1 was in that tumor tissues. Conclusion： Human UREB1 may be involved in the development of tumor and its tyrosine phosphorylation may affect the degree of tumor malignant.