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Reversible forskolin‐induced impairment of sucrase‐isomaltase mRNA levels, biosynthesis, and transport to the brush border membrane in Caco‐2 cells
Author(s) -
Rousset Monique,
Chantret Isabelle,
Darmoul Dalila,
Trugnan Germain,
Sapin Catherine,
Green Fiona,
Swallow Dallas,
Zweibaum Alain
Publication year - 1989
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1041410322
Subject(s) - sucrase , forskolin , brush border , enzyme , messenger rna , biosynthesis , biochemistry , biology , chemistry , membrane , gene , in vitro , vesicle
Abstract Hybridization analysis of mRNA with a cDNA probe for human sucrase‐isomaltase, pulse‐chase experiments with L‐[ 35 S]‐methionine followed by SDS‐PAGE, and immunofluorescence detection of sucrase‐isomaltase were used to analyze the level(s) at which forskolin interferes with the expression of the enzyme in Caco‐2 cells in culture. Three effects are observed in Caco‐2 cells treated with forskolin: (1) a marked decrease in the level of sucrase‐isomaltase mRNA, (2) a marked decrease in the biosynthesis of the enzyme without any alteration of its stability, and (3) an almost total inhibition of its transport to the brush border membrane. All three effects are reversible when the drug is removed from the culture medium, though this reversibility is asynchronous: transport to the brush border membrane resumes after 24 h, sucrase‐isomaltase mRNA levels are back to the normal after 5 days, whereas the biosynthesis of the enzyme, although increasing progressively, remains lower than in control cells, even 10 days after removal of the drug. The possibility that some effects are directly dependent on cAMP and others a consequence of changes in glucose metabolism is discussed.