Snail Regulates Cell-Matrix Adhesion by Regulation of the Expression of Integrins and Basement Membrane Proteins*
- Misako Haraguchi‡1,
- Tadashi Okubo‡,
- Yayoi Miyashita‡,
- Yasunori Miyamoto§,
- Masao Hayashi§,
- Tania N. Crotti¶,
- Kevin P. McHugh¶ and
- Masayuki Ozawa‡
- ‡Department of Biochemistry and Molecular Biology, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima 890-8544, Japan, the §Department of Biology, Ochanomizu University, Tokyo 112-8610, Japan, and the ¶Division of Rheumatology, Beth Israel Deaconess Medical Center, Harvard Medical School, Massachusetts 02115
- 1 To whom correspondence should be addressed. Tel.: 81-992-75-5246; Fax: 81-992-64-5618; E-mail: haraguci{at}m3.kufm.kagoshima-u.ac.jp.
Abstract
Snail, a transcriptional repressor of E-cadherin expression, plays a role in the process of epithelial-mesenchymal transition. However, the molecular basis of the role of snail in epithelial-mesenchymal transition has not been fully clarified. Here we show that the expression of snail in epithelial Madin-Darby canine kidney (MDCK) and A431 cells enhances both cell detachment and attachment. Snail did not confer resistance to anoikis induced by loss of contact but instead enhanced cell attachment to extracellular matrices such as fibronectin. This attachment was inhibited by Arg-Gly-Asp (RGD) peptides. Up-regulation of the promoter activity of integrin αV was observed in snail-expressing MDCK (MDCK/snail) cells. Snail also enhanced MDCK cell migration toward osteopontin that is a ligand for integrin αVβ3. We confirmed the reduction of basement membrane proteins such as laminin (LN) α3, β3, and γ2 (laminin-5/LN-5) and of receptors for LN-5 such as integrins α3, α6, or β4 in MDCK/snail or in snail-expressing A431 (A431/snail) cells. Nevertheless, suppression of LN-α3 chain by transient transfection of small interference RNAs resulted in no enhancement of cell detachment. We also found an induction of matrix metalloproteinase-3 in MDCK/snail and A431/snail cells. However, the inhibition of matrix metalloproteinase-3 showed no significant effect on the detachment of MDCK/snail cells. These results suggest that snail enhances cell detachment by multiple mechanism and leads to cell migration and reattachment at a second site, at least in part, by changing the expression of integrins in the cells.
Footnotes
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↵2 The abbreviations used are: EMT, epithelial-mesenchymal transition; ECM, extracellular matrix; LN, laminin; Col IV, collagen IV; FN, fibronectin; BM, basement membrane; MDCK, Madin-Darby canine kidney; DMEM, Dulbecco's modified Eagle's medium; FCS, fetal calf serum; PBS, phosphate-buffered saline; BSA, bovine serum albumin; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; RT, reverse transcription; siRNA, small interference RNA; poly-HEMA, poly-2-hydroxyethyl methacrylate; LDH, lactate dehydrogenase; CMV, cytomegalovirus; FL, firefly luciferase; RL, Renilla luciferase; PI, propidium iodide; CM, conditioned medium; DC, deoxycholic acid.
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↵* This work was supported by a grant-in-aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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- Received February 12, 2008.
- Revision received June 17, 2008.
- The American Society for Biochemistry and Molecular Biology, Inc.
