Regulation of Na+/glucose cotransporter (SGLT1) mRNA in LLC-PK1 cells.
The porcine kidney epithelial cell line LLC-PK1 expresses a sodium-coupled glucose cotransporter (SGLT1) together with other differentiation markers of renal proximal tubule such as trehalase and gamma-glutamyltranspeptidase. Expression is regulated by cell density and exogenous differentiation inducers such as hexamethylene bisacetamide (HMBA). Northern blot and PCR analysis of clonal cell populations indicated SGLT1 mRNA was not detectable in subconfluent cultures, but 2.2 and 3.9 kb SGLT1 mRNA species appeared after cell confluence, accompanying expression of the transport activity. SGLT1 mRNA levels were significantly increased after treatment of confluent cultures with HMBA, paralleling increases in the transport activity and immunodetectable 75 kD cotransporter subunit. SGLT1 mRNA was also increased after treatment of cultures with the cyclic AMP phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX), an inducer of Na+/glucose cotransport activity. The 3.9 kb SGLT1 transcript showed the largest increase after either HMBA or IBMX treatment. HMBA treatment also resulted in increased mRNA levels of two other differentiation markers--trehalase and gamma-glutamyltranspeptidase. By contrast, trehalase and gamma-glutamyltranspeptidase mRNA levels were not increased by IBMX. Regulation of Na+/glucose symporter expression by either cell density, cyclic AMP elevation, or differentiation inducer treatment occurs, at least in part, at the level of SGLT1 mRNA and can be dissociated from regulation of other differentiation markers.
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Department of Biochemistry and Molecular Biology, University of Texas Medical School, Houston 77225.
Kong
C T
CT
Peng
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Lever
J E
JE
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000081:Acetamides / Q000494:pharmacology; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001483:Base Sequence; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000235:genetics*; D002452:Cell Count; D002460:Cell Line; D004247:DNA / Q000032:analysis / Q000235:genetics; D004847:Epithelial Cells; D004848:Epithelium / Q000737:chemistry; D005786:Gene Expression Regulation; D007668:Kidney / Q000737:chemistry* / Q000166:cytology*; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D051269:Sodium-Glucose Transporter 1; D013552:Swine; D014198:Trehalase / Q000032:analysis / Q000235:genetics; D005723:gamma-Glutamyltransferase / Q000032:analysis / Q000235:genetics
D000081:Acetamides; D000970:Antineoplastic Agents; D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D012333:RNA, Messenger; D051269:Sodium-Glucose Transporter 1; D004247:DNA; D005723:gamma-Glutamyltransferase; D014198:Trehalase; C014026:hexamethylene bisacetamide; D015056:1-Methyl-3-isobutylxanthine