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Differentiation‐dependent induction of CYP1A1 in cultured rat small intestinal epithelial cells, colonocytes, and human colon carcinoma cells: Basement membrane‐mediated apoptosis
Author(s) -
Sterling Kenneth M.,
Cutroneo Kenneth R.
Publication year - 2002
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.10237
Subject(s) - matrigel , microbiology and biotechnology , transfection , cell culture , luciferase , chemistry , biology , in vitro , biochemistry , genetics
Rat small intestinal epithelial cells and human colon adenocarcinoma cells cultured on Matrigel™ expressed the differentiation specific enzyme, sucrase‐isomaltase, as determined by indirect immunofluorescence. Rat small intestinal epithelial cells, rat colonocytes, and human colon adenocarcinoma cells developed an altered morphology when cultured on Matrigel and became apoptotic within 24–48 h. Benzo[ a ]pyrene and 2,3,7,8‐tetrachlorodibenzo‐ p ‐dioxin caused a 2‐ and 5‐fold induction, respectively, of ethoxyresorufin‐ o‐ deethylase activity in rat small intestinal epithelial cells cultured on Matrigel. Benzo[ a ]pyrene‐ or 2,3,7,8‐tetrachlorodibenzo‐ p ‐dioxin‐induced ethoxyresorufin‐ o‐ deethylase activity in rat small intestinal epithelial cells cultured on plastic was not detected. 2,3,7,8‐tetrachlorodibenzo‐ p ‐dioxin treatment caused a 14‐fold induction of transfected, rat CYP1A1 ‐promoter‐luciferase activity in rat small intestinal epithelial cells cultured on Matrigel. Benzo[ a ]pyrene and 2,3,7,8‐tetrachlorodibenzo‐ p ‐dioxin treatment induced ethoxyresorufin‐ o‐ deethylase activity by 6‐ and 1.6‐fold, respectively in rat colonocytes cultured on Matrigel. Induction of ethoxyresorufin‐ o‐ deethylase activity was not observed in rat colonocytes cultured on plastic. CYP1A1 ‐promoter‐luciferase activity was induced 3‐fold by 2,3,7,8‐tetrachlorodibenzo‐ p ‐dioxin in rat colonocytes cultured on Matrigel. Induction of CYP1A1 ‐promoter‐luciferase activity in rat small intestinal epithelial cells or rat colonocytes cultured on plastic was not observed. Ethoxyresorufin‐ o‐ deethylase activity in human colon adenocarcinoma cells, cultured on either plastic or Matrigel, was induced 7‐fold by benzo[ a ]pyrene. 2,3,7,8‐Tetrachlorodibenzo‐ p ‐dioxin‐induced ethoxyresorufin‐ o‐ deethylase activity was 2‐fold greater in human colon adenocarcinoma cells cultured on Matrigel compared to cells cultured on plastic. Extracellular matrix‐mediated differentiation and apoptosis of intestinal cells provide in vitro systems for study of the regulation of CYP1A1 expression, carcinogen activation in the gut and mechanism(s) of apoptosis of colon cancer cells. J. Cell. Biochem. 86: 440–450, 2002. © 2002 Wiley‐Liss, Inc.