CFTR gene transfer corrects defective glycoconjugate secretion in human CF epithelial tracheal cells.
CFTR gene transfer corrects defective glycoconjugate secretion in human CF epithelial tracheal cells.
复制标题
CFTR 基因转移可纠正人 CF 上皮气管细胞中糖复合物分泌缺陷。
DOI：
10.1152/ajplung.1995.269.6.l855
复制
发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Paul,A
中科院分区：
文献类型：
--
作者：
Mergey,M;Lemnaouar,M;Veissiere,D;Perricaudet,M;Gruenert,DC;Picard,J;Capeau,J;Brahimi-Horn,MC;Paul,A
我们证明，在永生化正常人气管上皮细胞（NT-1 和 56FHTE8o-）中，14C 标记的糖缀合物分泌可能受到蛋白激酶 A (PKA) 和蛋白激酶 C (PKC) 信号通路激动剂的独立调节。相比之下，在永生化囊性纤维化 (CF) 人气管上皮细胞（CFT-1 和 CFT-2）中，PKA 特异性激动剂的调节存在缺陷，但 PKC 途径特异性激动剂的调节存在缺陷。为了表征囊性纤维化跨膜电导调节器（CFTR）在受调节的糖缀合物分泌中的参与，我们检查了腺病毒介导的 CFTR 基因转移到 CF 和对照细胞的效果。感染后48小时，在每个细胞50个空斑形成单位的感染复数下，通过逆转录聚合酶链式反应检测到高水平的CFTR mRNA，并通过免疫印迹证明CFTR蛋白的从头合成。基因转移至 CF 细胞不仅恢复了有缺陷的腺苷 3',5'-环单磷酸 (cAMP) 依赖性分泌，不仅恢复了氯化物的分泌，还恢复了糖缀合物的分泌。综上所述，这些结果证明了 CFTR 在 cAMP 介导的糖缀合物分泌中的作用。
We demonstrate that in immortalized normal human tracheal epithelial cells (NT-1 and 56FHTE8o-) 14C-labeled glycoconjugate secretion may be regulated independently by agonists of the protein kinase A (PKA) and protein kinase C (PKC) signaling pathways. In contrast, in immortalized cystic fibrosis (CF) human tracheal epithelial cells (CFT-1 and CFT-2), regulation is defective for agonists specific for the PKA but not for the PKC pathway. To characterize the involvement of the cystic fibrosis transmembrane conductance regulator (CFTR) in regulated glycoconjugate secretion, we examined the effect of adenovirus-mediated gene transfer of CFTR to CF and control cells. Forty-eight hours after infection, at a multiplicity of infection of 50 plaque-forming units per cell, high levels of CFTR mRNA were detected by reverse transcription-polymerase chain reaction, and de novo synthesis of CFTR protein was demonstrated by immunoblotting. Gene transfer to CF cells restored defective adenosine 3',5'-cyclic monophosphate (cAMP)-dependent secretion not only of chloride but also of glycoconjugates. Taken together, these results argue for a role for CFTR in cAMP-mediated glycoconjugate secretion.