title: Human intestinal epithelial cell line SK-CO15 is a new model system to study Na(+)/H(+) exchanger 3.
abstract: 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.affiliation: Division of Digestive Diseases, Department of Medicine, Emory University, Atlanta, GA 30322, USA.
authors.forename: Byong Kwon
authors.initials: BK
authors.lastname: Yoo
authors.forename: Murali Krishna
authors.initials: MK
authors.lastname: Yanda
authors.forename: Yi Ran
authors.initials: YR
authors.lastname: No
authors.forename: C Chris
authors.initials: CC
authors.lastname: Yun
mesh_terms: D027682:Cation Transport Proteins / Q000096:biosynthesis; D002460:Cell Line; D016764:Cell Polarity / Q000187:drug effects / Q000502:physiology; D005576:Colforsin / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D005938:Glucocorticoids / Q000494:pharmacology; D006801:Humans; D007413:Intestinal Mucosa / Q000187:drug effects / Q000502:physiology*; D008246:Lysophospholipids / Q000494:pharmacology; D010750:Phosphoproteins / Q000096:biosynthesis; D049368:Receptors, Lysophosphatidic Acid / Q000096:biosynthesis / Q000235:genetics; D000074082:Sodium-Hydrogen Exchanger 1; D000074083:Sodium-Hydrogen Exchanger 3; D017923:Sodium-Hydrogen Exchangers / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection
chemical_list: D027682:Cation Transport Proteins; D005938:Glucocorticoids; C512992:LPAR5 protein, human; D008246:Lysophospholipids; D010750:Phosphoproteins; D049368:Receptors, Lysophosphatidic Acid; C490998:SLC9A1 protein, human; C081517:SLC9A2 protein, human; C000613385:SLC9A3 protein, human; D000074082:Sodium-Hydrogen Exchanger 1; D000074083:Sodium-Hydrogen Exchanger 3; D017923:Sodium-Hydrogen Exchangers; C093376:sodium-hydrogen exchanger regulatory factor; D005576:Colforsin; D003907:Dexamethasone; C032881:lysophosphatidic acid