https://scholars.lib.ntu.edu.tw/handle/123456789/187940
標題: | 水管道蛋白於人體肺組織與細胞株之表現及調控 | 其他標題: | Expression And Regulation of Water Channel Proteins In Human Lung Tissues and Cell Lines | 作者: | 郭炳宏 | 關鍵字: | 水管道蛋白;人體肺組織與細胞株;基因表現及調控;Aquaporin;Water channels;Human lung tissues and cell line;Gene expression;Tissue distribution | 公開日期: | 1999 | 出版社: | 臺北市:國立臺灣大學醫學院內科 | 摘要: | 近年來細胞膜水管道(water channel)的發 現可說對細胞膜水滲透之分子機制提供 了新的解釋與研究方向。水管道是一膜蛋白族群,又稱為Aquaporins (AQP), 目前在老鼠已發現有八種水管道蛋白 (AQP1 至AQP8),人類水管道基因在人體 肺部組織之分部及表現仍未完全清楚。 吾人推測水管道蛋白或許與肺癌惡性胸 水之產生及某些cytokines, 如 Interleukin-2 (IL-2)等導致肺水腫之機制 有關。此研究之目的在於應用分子生物 學之技術檢視水管道蛋白在人體肺組織 及各種呼吸道細胞株之表現。並進一步 探討類固醇及IL-2 是否可誘導其表現。 我們選擇AQP1~5 基因設計PCR primer 。在細胞株方面,抽取RNA 再經 RT-PCR ,再經Northern blot 。在正常呼 吸道上皮細胞株(BSF-2)及肺腺癌細胞 株(A549 ,CRL-5806 ,CRL-5807)以及扁 平癌細胞株(HTB58)中只發現A549 細胞 有明顯AQP3 之表現。AQP3 基因可受類毋醇(dexamethasone 10-5 M 24 hr) 而調 控其表現,但在IL-2 (1 nM 8 hr)刺激下則 無明顯反應。在組織分部方面,我們收 集肺切除手術或胸水病患所得之肺部及 肋膜切片組織,運用水管道蛋白之多株抗體用於組織染色。發現AQP1 在肺部 各部位都有表現,AQP3 在遠端係支氣 管無表現,AQP5 則存於遠端係支氣管以下之肺組織。 Recent discovery of water channel proteins (aquaporins, AQP) has provided insight into the molecular mechanism of water transport across the cell membrane. Nevertheless, tissue distribution and the pathophysiology of these proteins in lung disorders remain unclear. This study applied techniques in molecular biology to screen human lung cell lines for the expression of known aquaporin genes, and also investigated tissue distribution of aquaporins in adult human lung and pleura by immunohistochemistry. Lung tissue sections, obtained during lobectomy, were analyzed by immunohistochemistry and immunoblot with aquaporin antibodies. Expression of AQP3 mRNA and protein is identified in A549 cells. No other cell lines of human airway epithelium and pleura in our laboratory were positive for other AQP genes. AQP3 mRNA expression in the dexamethasone-treated group (10 -5 M for 8 h) increased to approximately five times the level of that in the control group but did not change significantly after incubation with IL-2 (1 nM) for 8 hrs. Immunohistochemistry and Western blotting revealed that AQP1 is expressed in all portions of the lung, AQP3 is less strongly expressed in trachea and not detectable in distal lung. AQP5 is expressed in distal lung. Data obtained from his study might provide the basis for future study regarding the regulation and pathophysiology of aquaporins in human lung water homeostasis. |
URI: | http://ntur.lib.ntu.edu.tw//handle/246246/23420 | 其他識別: | 882314B002229 | Rights: | 國立臺灣大學醫學院內科 |
顯示於: | 醫學系 |
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882314B002229.pdf | 34.43 kB | Adobe PDF | 檢視/開啟 |
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