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The DHE cell line as a model for studying rat gastro-intestinal mucin expression: effects of dexamethasone.
- Source :
-
European journal of cell biology [Eur J Cell Biol] 2004 Aug; Vol. 83 (7), pp. 347-58. - Publication Year :
- 2004
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Abstract
- The expression of mucin genes was evaluated in rat intestinal cell lines in order to establish an in vitro model for investigating the regulation of intestinal mucin expression in this species. Two rat intestinal cancer cell lines (DHE, LGA) and three nontumoral rat intestinal cell lines (IEC6, IEC17, IEC18) were screened. The mRNA expression of rMuc1, rMuc2, rMuc3, rMuc4, and rMuc5AC mucin genes was studied by semiquantitative RT-PCR, real-time RT-PCR and Northern-blot analysis. Results were correlated with immunohistochemical expression of rat gastric and intestinal mucin proteins, and secretion of glycoconjugates was examined by enzyme-linked lectin assay. We showed that mRNA of rMucl and rMuc2 were constitutively expressed in all IEC cell populations but periodic acid Schiff staining of these cells did not reveal the presence of glycoproteins. DHE cells expressed rMuc1-5AC mRNA and LGA expressed the same mucins but the level of rMuc4 was much lower. Mucin mRNA expression also differed in relation with the length of cultivation. Immunocytochemical studies revealed the presence of gastric and intestinal mucins in the two tumoral cell lines. Functional experiments showed that bethanechol, A23187 and PMA stimulated release of glycoconjugates in DHE but not in LGA cells. Treatment of DHE cells with dexamethasone (10(-7) mol/l) enhanced rMuc2 mRNA but decreased rMuc1 and rMuc5AC mRNA. Real-time RT-PCR showed that the expression of rMuc1 and rMuc5AC genes was reduced by more than tenfold after 24 h. The increased expression of rMuc2 gene was confirmed by Northern blot analysis. In conclusion, DHE cells provide a valuable cellular model for research on rat mucin secretion and expression.
- Subjects :
- Animals
Bethanechol pharmacology
Blotting, Northern
Calcimycin pharmacology
Carcinogens pharmacology
Gastric Mucosa metabolism
Gastric Mucosa pathology
Immunohistochemistry
Intestinal Neoplasms metabolism
Intestinal Neoplasms pathology
Ionophores pharmacology
Muscarinic Agonists pharmacology
RNA, Messenger biosynthesis
Rats
Reverse Transcriptase Polymerase Chain Reaction
Species Specificity
Tetradecanoylphorbol Acetate pharmacology
Anti-Inflammatory Agents pharmacology
Cell Line, Tumor metabolism
Cell Line, Tumor pathology
Dexamethasone pharmacology
Gastric Mucins biosynthesis
Gene Expression Regulation, Neoplastic drug effects
Subjects
Details
- Language :
- English
- ISSN :
- 0171-9335
- Volume :
- 83
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- European journal of cell biology
- Publication Type :
- Academic Journal
- Accession number :
- 15503858
- Full Text :
- https://doi.org/10.1078/0171-9335-00391