Cozens A L, Yezzi M J, Chin L, Simon E M, Finkbeiner W E, Wagner J A, Gruenert D C
Cardiovascular Research Institute, University of California, San Francisco 94143.
Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5171-5. doi: 10.1073/pnas.89.11.5171.
Tracheobronchial glands were isolated and cultured from a patient with cystic fibrosis (CF). Cultured epithelial cells were transformed with pSVori-. All transformed cell lines express cytokeratin filaments and at early passages express the junctional complex molecule cell CAM 120/80, indicating their epithelial origin. Several gland cell lines express antigens that localize to secretory cells in vivo. Cl- transport measured by 36Cl efflux shows that CF gland epithelial cells, like CF surface airway and nasal polyp epithelial cells, are unable to respond to increases in intracellular cAMP. However, they do produce an increase in intracellular cAMP after treatment with isoproterenol or forskolin. One CF gland cell line shows increased intracellular calcium in response to a number of agents and increased Cl- efflux comparable to that observed in a non-CF airway surface epithelial cell line after addition of calcium ionophore. All cell lines express CF transmembrane conductance regulator mRNA, as measured by PCR amplification of first-strand cDNA. The CF tracheobronchial gland cell lines described here are compound heterozygotes, having a single copy of the delta F508 mutation.
从一名囊性纤维化（CF）患者身上分离并培养气管支气管腺。培养的上皮细胞用pSVori - 进行转化。所有转化细胞系均表达细胞角蛋白丝，并且在早期传代时表达连接复合体分子细胞黏附分子120/80，表明其上皮起源。几个腺细胞系表达在体内定位于分泌细胞的抗原。通过³⁶Cl外流测量的Cl⁻转运表明，CF腺上皮细胞与CF气道表面和鼻息肉上皮细胞一样，无法对细胞内cAMP的增加做出反应。然而，在用异丙肾上腺素或福斯高林处理后，它们确实会使细胞内cAMP增加。一种CF腺细胞系对多种试剂的反应显示细胞内钙增加，并且在添加钙离子载体后，其Cl⁻外流增加，与在非CF气道表面上皮细胞系中观察到的情况相当。通过对第一链cDNA进行PCR扩增测量，所有细胞系均表达CF跨膜电导调节因子mRNA。这里描述的CF气管支气管腺细胞系是复合杂合子，具有单个δF508突变拷贝。
