Mitogen-activated protein kinase activation regulates intestinal epithelial differentiation

D. Taupin, D. K. Podolsky

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

Background and Aims: The human colon cancer-derived cell line HT29 displays a multipotent phenotype. A subclone of HT29 cells containing numerous mucous granules and termed HT29-18-N2 was studied to determine the cellular mechanisms underlying a switch to the differentiated phenotype. Methods: Northern (RNA) blotting, immunoblotting, and immunocytochemistry of HT29-N2 cells, grown under glucose-containing and glucose-free conditions with or without the use of the mitogen-activated protein (MAP) kinase kinase (MEK) inhibitor PD98059, were performed. Results: Loss of activation of the MAP kinases ERK1 and ERK2 in HT29-N2 cells upon a change to glucose-free growth medium preceded the change in phenotype and upregulation of the goblet cell gene product intestinal trefoil factor (ITF). Long-term pharmacological MAP kinase inhibition with the MEK inhibitor PD98059 induced expression of the terminal differentiation markers ITF, sucrase-isomaltase, and the mucin gene MUC2. This was accompanied by morphological evidence of gland formation and mucin secretion and the appearance of discrete goblet cell and enterocyte populations. Induction of ITF and sucrase-isomaltase after MEK inhibition in HT29-N2 cells did not involve loss of MAP kinase responsiveness and was not mediated by receptor tyrosine kinases. Conclusions: Regulation of ERK activation may be a key biochemical switch responsible for terminal differentiation of components of the crypt-villus unit.

Original languageEnglish (US)
Pages (from-to)1072-1080
Number of pages9
JournalGastroenterology
Volume116
Issue number5
DOIs
StatePublished - 1999

Fingerprint

HT29 Cells
Mitogen-Activated Protein Kinases
Mitogen-Activated Protein Kinase Kinases
Oligo-1,6-Glucosidase
Sucrase
Goblet Cells
Mucins
Phenotype
Glucose
Enterocytes
Differentiation Antigens
Receptor Protein-Tyrosine Kinases
Immunoblotting
Northern Blotting
Colonic Neoplasms
Genes
Up-Regulation
Immunohistochemistry
Pharmacology
RNA

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Mitogen-activated protein kinase activation regulates intestinal epithelial differentiation. / Taupin, D.; Podolsky, D. K.

In: Gastroenterology, Vol. 116, No. 5, 1999, p. 1072-1080.

Research output: Contribution to journalArticle

@article{ab9073d3e0524c0eb70fb277976f96d6,
title = "Mitogen-activated protein kinase activation regulates intestinal epithelial differentiation",
abstract = "Background and Aims: The human colon cancer-derived cell line HT29 displays a multipotent phenotype. A subclone of HT29 cells containing numerous mucous granules and termed HT29-18-N2 was studied to determine the cellular mechanisms underlying a switch to the differentiated phenotype. Methods: Northern (RNA) blotting, immunoblotting, and immunocytochemistry of HT29-N2 cells, grown under glucose-containing and glucose-free conditions with or without the use of the mitogen-activated protein (MAP) kinase kinase (MEK) inhibitor PD98059, were performed. Results: Loss of activation of the MAP kinases ERK1 and ERK2 in HT29-N2 cells upon a change to glucose-free growth medium preceded the change in phenotype and upregulation of the goblet cell gene product intestinal trefoil factor (ITF). Long-term pharmacological MAP kinase inhibition with the MEK inhibitor PD98059 induced expression of the terminal differentiation markers ITF, sucrase-isomaltase, and the mucin gene MUC2. This was accompanied by morphological evidence of gland formation and mucin secretion and the appearance of discrete goblet cell and enterocyte populations. Induction of ITF and sucrase-isomaltase after MEK inhibition in HT29-N2 cells did not involve loss of MAP kinase responsiveness and was not mediated by receptor tyrosine kinases. Conclusions: Regulation of ERK activation may be a key biochemical switch responsible for terminal differentiation of components of the crypt-villus unit.",
author = "D. Taupin and Podolsky, {D. K.}",
year = "1999",
doi = "10.1016/S0016-5085(99)70010-7",
language = "English (US)",
volume = "116",
pages = "1072--1080",
journal = "Gastroenterology",
issn = "0016-5085",
publisher = "W.B. Saunders Ltd",
number = "5",

}

TY - JOUR

T1 - Mitogen-activated protein kinase activation regulates intestinal epithelial differentiation

AU - Taupin, D.

AU - Podolsky, D. K.

PY - 1999

Y1 - 1999

N2 - Background and Aims: The human colon cancer-derived cell line HT29 displays a multipotent phenotype. A subclone of HT29 cells containing numerous mucous granules and termed HT29-18-N2 was studied to determine the cellular mechanisms underlying a switch to the differentiated phenotype. Methods: Northern (RNA) blotting, immunoblotting, and immunocytochemistry of HT29-N2 cells, grown under glucose-containing and glucose-free conditions with or without the use of the mitogen-activated protein (MAP) kinase kinase (MEK) inhibitor PD98059, were performed. Results: Loss of activation of the MAP kinases ERK1 and ERK2 in HT29-N2 cells upon a change to glucose-free growth medium preceded the change in phenotype and upregulation of the goblet cell gene product intestinal trefoil factor (ITF). Long-term pharmacological MAP kinase inhibition with the MEK inhibitor PD98059 induced expression of the terminal differentiation markers ITF, sucrase-isomaltase, and the mucin gene MUC2. This was accompanied by morphological evidence of gland formation and mucin secretion and the appearance of discrete goblet cell and enterocyte populations. Induction of ITF and sucrase-isomaltase after MEK inhibition in HT29-N2 cells did not involve loss of MAP kinase responsiveness and was not mediated by receptor tyrosine kinases. Conclusions: Regulation of ERK activation may be a key biochemical switch responsible for terminal differentiation of components of the crypt-villus unit.

AB - Background and Aims: The human colon cancer-derived cell line HT29 displays a multipotent phenotype. A subclone of HT29 cells containing numerous mucous granules and termed HT29-18-N2 was studied to determine the cellular mechanisms underlying a switch to the differentiated phenotype. Methods: Northern (RNA) blotting, immunoblotting, and immunocytochemistry of HT29-N2 cells, grown under glucose-containing and glucose-free conditions with or without the use of the mitogen-activated protein (MAP) kinase kinase (MEK) inhibitor PD98059, were performed. Results: Loss of activation of the MAP kinases ERK1 and ERK2 in HT29-N2 cells upon a change to glucose-free growth medium preceded the change in phenotype and upregulation of the goblet cell gene product intestinal trefoil factor (ITF). Long-term pharmacological MAP kinase inhibition with the MEK inhibitor PD98059 induced expression of the terminal differentiation markers ITF, sucrase-isomaltase, and the mucin gene MUC2. This was accompanied by morphological evidence of gland formation and mucin secretion and the appearance of discrete goblet cell and enterocyte populations. Induction of ITF and sucrase-isomaltase after MEK inhibition in HT29-N2 cells did not involve loss of MAP kinase responsiveness and was not mediated by receptor tyrosine kinases. Conclusions: Regulation of ERK activation may be a key biochemical switch responsible for terminal differentiation of components of the crypt-villus unit.

UR - http://www.scopus.com/inward/record.url?scp=0032960673&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032960673&partnerID=8YFLogxK

U2 - 10.1016/S0016-5085(99)70010-7

DO - 10.1016/S0016-5085(99)70010-7

M3 - Article

VL - 116

SP - 1072

EP - 1080

JO - Gastroenterology

JF - Gastroenterology

SN - 0016-5085

IS - 5

ER -