PMID- 25263207
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20140929
LR  - 20211021
IS  - 2051-817X (Print)
IS  - 2051-817X (Electronic)
IS  - 2051-817X (Linking)
VI  - 2
IP  - 9
DP  - 2014 Sep 1
TI  - HEK-293 cells expressing the cystic fibrosis transmembrane conductance regulator 
      (CFTR): a model for studying regulation of Cl- transport.
LID - 10.14814/phy2.12158 [doi]
LID - e12158
AB  - The Human Embryonic Kidney 293 cell line (HEK-293) readily lends itself to 
      genetic manipulation and is a common tool for biologists to overexpress proteins 
      of interest and study their function and molecular regulation. Although these 
      cells have some limitations, such as an inability to form resistive monolayers 
      necessary for studying transepithelial ion transport, they are nevertheless 
      valuable in studying individual epithelial ion transporters. We report the use of 
      HEK-293 cells to study the cystic fibrosis transmembrane conductance regulator 
      (CFTR) Cl(-) channel. While HEK-293 cells endogenously express mRNA for the Cl(-) 
      channels, ClC-2 and TMEM16A, they neither express CFTR mRNA nor protein. 
      Therefore, we stably transfected HEK-293 cells with EGFP-CFTR (HEK-CFTR) and 
      demonstrated CFTR function by measuring forskolin-stimulated iodide efflux. This 
      efflux was inhibited by CFTRinh172, and the protein kinase A inhibitor H89, but 
      not by Ca(2+) chelation. In contrast to intestinal epithelia, forskolin 
      stimulation does not increase surface CFTR expression and does not require intact 
      microtubules in HEK-CFTR. To investigate the role of an endogenous GalphaS-coupled 
      receptor, we examined the bile acid receptor, TGR5. Although HEK-CFTR cells 
      express TGR5, the potent TGR5 agonist lithocholic acid (LCA; 5-500 mumol/L) did 
      not activate CFTR. Furthermore, forskolin, but not LCA, increased [cAMP]i in 
      HEK-CFTR suggesting that endogenous TGR5 may not be functionally linked to GalphaS. 
      However, LCA did increase [Ca(2+)]i and interestingly, abolished 
      forskolin-stimulated iodide efflux. Thus, we propose that the stable HEK-CFTR 
      cell line is a useful model to study the multiple signaling pathways that 
      regulate CFTR.
CI  - (c) 2014 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on 
      behalf of the American Physiological Society and The Physiological Society.
FAU - Domingue, Jada C
AU  - Domingue JC
AD  - Department of Physiology and Biophysics, University of Illinois at Chicago, 
      Chicago, Illinois.
FAU - Ao, Mei
AU  - Ao M
AD  - Department of Physiology and Biophysics, University of Illinois at Chicago, 
      Chicago, Illinois.
FAU - Sarathy, Jayashree
AU  - Sarathy J
AD  - Department of Physiology and Biophysics, University of Illinois at Chicago, 
      Chicago, Illinois Department of Biological Sciences, Benedictine University, 
      Lisle, Illinois.
FAU - George, Alvin
AU  - George A
AD  - Department of Physiology and Biophysics, University of Illinois at Chicago, 
      Chicago, Illinois.
FAU - Alrefai, Waddah A
AU  - Alrefai WA
AD  - Department of Medicine, University of Illinois at Chicago, Chicago, Illinois 
      Jesse Brown V.A. Medical Center, Chicago, Illinois.
FAU - Nelson, Deborah J
AU  - Nelson DJ
AD  - Department of Pharmacological and Physiological Sciences, The University of 
      Chicago, Chicago, Illinois.
FAU - Rao, Mrinalini C
AU  - Rao MC
AD  - Department of Physiology and Biophysics, University of Illinois at Chicago, 
      Chicago, Illinois Department of Medicine, University of Illinois at Chicago, 
      Chicago, Illinois.
LA  - eng
GR  - I01 BX000152/BX/BLRD VA/United States
GR  - IK6 BX005243/BX/BLRD VA/United States
GR  - R01 DK071596/DK/NIDDK NIH HHS/United States
GR  - R01 GM036823/GM/NIGMS NIH HHS/United States
PT  - Journal Article
DEP - 20140928
PL  - United States
TA  - Physiol Rep
JT  - Physiological reports
JID - 101607800
PMC - PMC4270233
OTO - NOTNLM
OT  - CFTR
OT  - Cl- transport
OT  - HEK-293
OT  - TGR5
EDAT- 2014/09/30 06:00
MHDA- 2014/09/30 06:01
PMCR- 2014/09/28
CRDT- 2014/09/30 06:00
PHST- 2014/09/30 06:00 [entrez]
PHST- 2014/09/30 06:00 [pubmed]
PHST- 2014/09/30 06:01 [medline]
PHST- 2014/09/28 00:00 [pmc-release]
AID - 2/9/e12158 [pii]
AID - phy212158 [pii]
AID - 10.14814/phy2.12158 [doi]
PST - epublish
SO  - Physiol Rep. 2014 Sep 28;2(9):e12158. doi: 10.14814/phy2.12158. Print 2014 Sep 1.