EXPRESSION OF NORMAL AND CYSTIC-FIBROSIS PHENOTYPES BY CONTINUOUS AIRWAY EPITHELIAL-CELL LINES
EXPRESSION OF NORMAL AND CYSTIC-FIBROSIS PHENOTYPES BY CONTINUOUS AIRWAY EPITHELIAL-CELL LINES
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DOI：
10.1152/ajplung.1990.259.6.l496
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发表时间：
1990-12-01
影响因子：
--
通讯作者：
WELSH, MJ
中科院分区：
文献类型：
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作者：
JEFFERSON, DM;VALENTICH, JD;WELSH, MJ
囊性纤维化（CF）和正常对照个体的连续上皮细胞系需要了解CF的遗传和细胞缺陷。 我们用逆转录病毒将SV 40大T抗原转染到鼻上皮细胞中。 转化的连续细胞系被分离，表达上皮标记物，细胞角蛋白，和紧密连接。 北方印迹分析表明，所有的细胞系表达推定的CF基因mRNA。 对跨上皮电解质转运的研究表明，CF和正常细胞系产生跨上皮电阻。 正常但非CF细胞系分泌Cl-，以响应增加细胞内腺苷3 '，5'-环一磷酸（cAMP）水平的激动剂（异丙肾上腺素、毛喉素和cAMP的膜渗透类似物）或响应激活蛋白激酶C的促肿瘤佛波酯。 与此相反，Ca 2+升高激动剂缓激肽和Ca 2+离子载体A23187刺激分泌在正常和CF细胞系。 我们已经产生的连续细胞系保持其适当的表型，并将作为有用的工具，在理解CF的病理生理。
Continuous epithelial cell lines from individuals with cystic fibrosis (CF) and normal controls are required to understand the genetic and cellular defects in CF. We used retroviruses to transduce SV40 large T antigen into nasal epithelial cells. Transformed continuous cell lines were isolated that expressed epithelial markers, cytokeratin, and tight junctions. Northern blot analysis shows that all of the cell lines express the putative CF gene mRNA. Studies of transepithelial electrolyte transport show that CF and normal cell lines develop a transepithelial electrical resistance. Normal but not CF cell lines secreted Cl- in response to agonists that increase cellular levels of adenosine 3',5'-cyclic monophosphate (cAMP) (isoproterenol, forskolin, and a membrane-permeant analogue of cAMP) or in response to a tumor-promoting phorbol ester that activates protein kinase C. In contrast, the Ca2+ -elevating agonist bradykinin and the Ca2+ ionophore A23187 stimulated secretion in both normal and CF cell lines. The continuous cell lines we have produced maintain their proper phenotypes and will serve as useful tools in understanding the pathophysiology of CF.