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Lumican inhibits cell migration through alpha 2 beta 1 integrin
Tekijät: Zeltz C, Brezillon S, Kapyla J, Eble JA, Bobichon H, Terryn C, Perreau C, Franz CM, Heino J, Maquart FX, Wegrowski Y
Kustantaja: ELSEVIER INC
Julkaisuvuosi: 2010
Journal: Experimental Cell Research
Tietokannassa oleva lehden nimi: EXPERIMENTAL CELL RESEARCH
Lehden akronyymi: EXP CELL RES
Numero sarjassa: 17
Vuosikerta: 316
Numero: 17
Aloitussivu: 2922
Lopetussivu: 2931
Sivujen määrä: 10
ISSN: 0014-4827
DOI: https://doi.org/10.1016/j.yexcr.2010.08.002
Tiivistelmä
Lumican, an extracellular matrix protein of the small leucine-rich proteoglycan family, has been shown to impede melanoma progression by inhibiting cell migration. In the present study, we show that lumican targets alpha 2 beta 1 integrin thereby inhibiting cell migration. A375 melanoma cells were transfected with siRNA directed against the alpha 2 integrin subunit. Compared to A375 control cells, the anti-migratory effect of lumican was abrogated on transfected A375 cells. Moreover, lumican inhibited the chemotactic migration of Chinese hamster ovary (CHO) cells stably transfected with a2 integrin subunit (CHO-A2) but not that of wild-type CHO cells (CHO-WT) lacking this subunit. In contrast to CHO-WT cells, we observed in time-lapse microscopy a decrease of CHO-A2 cell migration speed in presence of lumican. Focal adhesion kinase phosphorylated at tyrosine-397 (pFAK) and total FAR were analysed in CHO-WT and CHO-A2 cells. A significant decrease of the ratio pFAK/FAK was shown in presence of recombinant human lumican. Using solid phase assays, a direct binding between lumican and the alpha 2 beta 1 integrin was demonstrated. This interaction did not involve the glycan moiety of lumican and was cation independent. Lumican was also able to bind the activated I domain of the alpha 2 integrin subunit with a K(d) >= 200 nM. In conclusion, we demonstrated for the first time that the inhibition of cell migration by lumican depends on a direct binding between the core protein of lumican and the alpha 2 beta 1 integrin. (C) 2010 Elsevier Inc. All rights reserved.
Lumican, an extracellular matrix protein of the small leucine-rich proteoglycan family, has been shown to impede melanoma progression by inhibiting cell migration. In the present study, we show that lumican targets alpha 2 beta 1 integrin thereby inhibiting cell migration. A375 melanoma cells were transfected with siRNA directed against the alpha 2 integrin subunit. Compared to A375 control cells, the anti-migratory effect of lumican was abrogated on transfected A375 cells. Moreover, lumican inhibited the chemotactic migration of Chinese hamster ovary (CHO) cells stably transfected with a2 integrin subunit (CHO-A2) but not that of wild-type CHO cells (CHO-WT) lacking this subunit. In contrast to CHO-WT cells, we observed in time-lapse microscopy a decrease of CHO-A2 cell migration speed in presence of lumican. Focal adhesion kinase phosphorylated at tyrosine-397 (pFAK) and total FAR were analysed in CHO-WT and CHO-A2 cells. A significant decrease of the ratio pFAK/FAK was shown in presence of recombinant human lumican. Using solid phase assays, a direct binding between lumican and the alpha 2 beta 1 integrin was demonstrated. This interaction did not involve the glycan moiety of lumican and was cation independent. Lumican was also able to bind the activated I domain of the alpha 2 integrin subunit with a K(d) >= 200 nM. In conclusion, we demonstrated for the first time that the inhibition of cell migration by lumican depends on a direct binding between the core protein of lumican and the alpha 2 beta 1 integrin. (C) 2010 Elsevier Inc. All rights reserved.
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