CFTR in K562 human leukemic cells
CFTR in K562 human leukemic cells
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DOI：
10.1152/ajpcell.00320.2002
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发表时间：
2003-08-01
影响因子：
5.5
通讯作者：
Kotsias, BA
中科院分区：
文献类型：
--
作者：
Assef, YA;Damiano, AE;Kotsias, BA
In this study, the expression and functional characterization of CFTR ( cystic fibrosis transmembrane regulator) was determined in K562 chronic human leukemia cells. Expression of the CFTR gene product was determined by RT-PCR and confirmed by immunohistochemistry and Western blot analysis. Functional characterization of CFTR Cl- channel activity was conducted with patch-clamp techniques. Forskolin, an adenylyl cyclase activator, induced an anion-selective channel with a linear current-voltage relationship and a single-channel conductance of 11 pS. This cAMP-activated channel had a P-gluconate/P-Cl or P-F/P-Cl perm-selectivity ratio of 0.35 and 0.30, respectively, and was inhibited by the CFTR blocker glibenclamide and the anti-CFTR antibody MAb 13-1, when added to the cytoplasmatic side of the patch. Glibenclamide decreased the open probability increasing the frequency of open-to-closed transitions. Addition of 200 muM DIDS caused an irreversible block of the channels when added to the cytosolic side of inside-out patches. These and other observations indicate a widespread distribution of CFTR gene expression and suggest that this channel protein may function in most human cells to help maintain cellular homeostasis.
