Promoter-independent regulation of vimentin expression in mammary epithelial cells by val(12)ras and TGFbeta.
The 1,029 series of mammary epithelial cell lines (D6, GP+E, r3 and r3T) are progressively more transformed: the latter two by val(12)ras. These cell lines respond to TGFbeta by undergoing early events of epithelial-mesenchymal transition (EMT), including morphological changes and redistribution of E-cadherin. Tumors formed by r3T cells in the choroid of the eye express vimentin, a late marker of EMT, possibly in response to TGFbeta. In vitro, vimentin expression is induced in all the cell lines by TGFbeta treatment, whereas cytokeratin expression is only slightly affected. Surprisingly, ras transformation results in a 10-fold suppression of vimentin expression. Neither suppression of vimentin by ras transformation nor induction by TGFbeta is mediated by the vimentin promoter in r3T cells. In transient transfection assays, several human vimentin promoter constructs are more active in the low-expressing r3T cell line than in the vimentin-expressing mesenchymal cell line NIH3T3. In the r3T cells, there is no effect of TGFbeta treatment for 9 days on the activity of either promoter. Azacytidine treatment does not affect vimentin expression in either NIH3T3 or r3T, suggesting that promoter methylation is not the mechanism of suppression by ras. Finally, the half-life of the vimentin mRNA is similar in both the r3T cells and NIH3T3 cells. We conclude that the suppression of vimentin expression by ras, and the relief of this suppression by TGFbeta, occurs in a promoter-independent fashion, possibly through sequences in the first or second intron.
Yates
Bradley
B
The Forsyth Institute, 140 The Fenway, Boston, MA 02115, USA.
Zetterberg
Craig
C
Rajeev
Vaishali
V
Reiss
Michael
M
Rittling
Susan R
SR
D000818:Animals; D002461:Cell Line, Transformed; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D018507:Gene Expression Regulation, Developmental; D020868:Gene Silencing; D006801:Humans; D053538:Keratin-18 / Q000235:genetics / Q000378:metabolism; D008648:Mesoderm / Q000378:metabolism / Q000473:pathology; D051379:Mice; D009362:Neoplasm Metastasis / Q000235:genetics* / Q000473:pathology; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics; D016212:Transforming Growth Factor beta / Q000378:metabolism* / Q000494:pharmacology; D014746:Vimentin / Q000235:genetics*; D018631:ras Proteins / Q000235:genetics / Q000378:metabolism*
D053538:Keratin-18; D016212:Transforming Growth Factor beta; D014746:Vimentin; D018631:ras Proteins
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