The DHE cell line as a model for studying rat gastro-intestinal mucin expression: effects of dexamethasone.
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.
Trompette
Aurélien
A
INSERM U45, IFR62, Faculté de médecine Laënnec, Lyon, France.
Blanchard
Carine
C
Zoghbi
Sandra
S
Bara
Jacques
J
Claustre
Jean
J
Jourdan
Gérard
G
Chayvialle
Jean Alain
JA
Plaisancé
Pascale
P
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D018723:Bethanechol / Q000494:pharmacology; D015152:Blotting, Northern; D000001:Calcimycin / Q000494:pharmacology; D002273:Carcinogens / Q000494:pharmacology; D045744:Cell Line, Tumor / Q000378:metabolism / Q000473:pathology; D003907:Dexamethasone / Q000494:pharmacology*; D005752:Gastric Mucins / Q000096:biosynthesis*; D005753:Gastric Mucosa / Q000378:metabolism / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D007150:Immunohistochemistry; D007414:Intestinal Neoplasms / Q000378:metabolism / Q000473:pathology; D007476:Ionophores / Q000494:pharmacology; D018721:Muscarinic Agonists / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013045:Species Specificity; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000893:Anti-Inflammatory Agents; D002273:Carcinogens; D005752:Gastric Mucins; D007476:Ionophores; D018721:Muscarinic Agonists; D012333:RNA, Messenger; D018723:Bethanechol; D000001:Calcimycin; D003907:Dexamethasone; D013755:Tetradecanoylphorbol Acetate