Document Detail


Human intestinal epithelial cell line SK-CO15 is a new model system to study Na(+)/H(+) exchanger 3.
MedLine Citation:
PMID:  22556145     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The Caco-2 cell line represents absorptive polarized intestinal epithelial cells that express multiple forms of Na(+)/H(+) exchanger (NHE) in their plasma membranes. Caco-2 cells express the major apical NHE isoform NHE3, but low NHE3 expression together with inefficient transfection often hamper intended studies. In this study, we examined whether SK-CO15 cells could be used to study NHE3 regulation. SK-CO15 cells grown on Transwell inserts developed polarized epithelial cells with microvilli. The transfection efficiency of SK-CO15 cells was markedly higher compared with Caco-2 cells, an advantage in gene transfer and knockout. SK-CO15 cells expressed NHE1, NHE2, and NHE3. NHE3 expression was significantly greater in these cells than Caco-2, and NHE3 comprised more than half of total NHE activity. Apical expression of NHE3 in SK-CO15 cells was confirmed by confocal immunofluorescence and surface biotinylation. NHE regulatory factors NHERF1 and NHERF2, which are important for regulation of NHE3 activity, were expressed in these cells. Stimulatory response of NHE3 in SK-CO15 cells was assessed by dexamethasone and lysophosphatidic acid (LPA). Treatment with dexamethasone for 24-48 h increased NHE3 expression and activity. Similarly to Caco-2 cells, SK-CO15 cells lacked the expression of the LPA receptor LPA(5,) but exogenous expression of LPA(5) resulted in acute stimulation of NHE3. Forskolin acutely inhibited NHE3 activity in SK-CO15 cells, further attesting the validity of these cells. We conclude that SK-CO15 cells with the amenity for transfection and high endogenous NHE3 expression are a new and better cell model for NHE3 regulatory investigation than widely used Caco-2 cells.
Authors:
Byong Kwon Yoo; Murali Krishna Yanda; Yi Ran No; C Chris Yun
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2012-05-03
Journal Detail:
Title:  American journal of physiology. Gastrointestinal and liver physiology     Volume:  303     ISSN:  1522-1547     ISO Abbreviation:  Am. J. Physiol. Gastrointest. Liver Physiol.     Publication Date:  2012 Jul 
Date Detail:
Created Date:  2012-07-16     Completed Date:  2012-10-03     Revised Date:  2014-05-02    
Medline Journal Info:
Nlm Unique ID:  100901227     Medline TA:  Am J Physiol Gastrointest Liver Physiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  G180-8     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Cation Transport Proteins / biosynthesis
Cell Line
Cell Polarity / drug effects,  physiology
Colforsin / pharmacology
Dexamethasone / pharmacology
Glucocorticoids / pharmacology
Humans
Intestinal Mucosa / drug effects,  physiology*
Lysophospholipids / pharmacology
Phosphoproteins / biosynthesis
Receptors, Lysophosphatidic Acid / biosynthesis,  genetics
Sodium-Hydrogen Antiporter / biosynthesis*,  genetics
Transfection
Grant Support
ID/Acronym/Agency:
DK-061418/DK/NIDDK NIH HHS; DK-061418S1/DK/NIDDK NIH HHS; DK-064399/DK/NIDDK NIH HHS; R01 DK061418/DK/NIDDK NIH HHS; T32 DK007771/DK/NIDDK NIH HHS; T32-DK-007771/DK/NIDDK NIH HHS
Chemical
Reg. No./Substance:
0/Cation Transport Proteins; 0/Glucocorticoids; 0/LPAR5 protein, human; 0/Lysophospholipids; 0/Phosphoproteins; 0/Receptors, Lysophosphatidic Acid; 0/SLC9A1 protein, human; 0/SLC9A2 protein, human; 0/Sodium-Hydrogen Antiporter; 0/sodium-hydrogen exchanger 3; 0/sodium-hydrogen exchanger regulatory factor; 1F7A44V6OU/Colforsin; 22002-87-5/lysophosphatidic acid; 7S5I7G3JQL/Dexamethasone
Comments/Corrections

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