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Hong, Kun-Jing; Wu, Deng-Chyang; Cheng, Kuang-Hung; Chen, Li-Tzong; Hung, Wen-Chun
Journal of cellular physiology, February 2014, Letnik: 229, Številka: 2Journal Article
The Reversion‐inducing Cysteine‐rich Protein with Kazal Motifs (RECK) gene encodes a membrane‐anchored glycoprotein that exhibits strong inhibitory activity against various matrix metalloproteinases (MMPs) and a disintegrin and metalloproteinase 10 (ADAM10). RECK functions as a tumor suppressor by inhibiting migration, invasion, and angiogenesis. However, whether RECK can modulate the stem‐like phenotypes of cancer cells is not known. In this study, we demonstrate that RECK is down‐regulated in gastric cancer cells and is further reduced in CD133‐positive cancer stem‐like cells. Ectopic expression of RECK induces down‐regulation of the expression of stemness genes including Sox2, Oct4, and Nanog and the cancer stem cell marker CD133. Treatment of DAPT (a γ‐secretase inhibitor) or TAPI‐2 (a hydroxamate‐based inhibitor of MMPs, tumor necrosis factor α converting enzyme and ADAM17) reduces Notch1 shedding and activation which results in attenuation of stemness genes and CD133. Our data show that ADAM10 and ADAM17 are co‐pulled down by RECK suggesting a physical interaction between RECK and ADAMs on cell surface. In addition, RECK suppresses sphere formation and sphere size of CD133‐positive gastric cancer cells. Overexpression of Notch intracellular domain (NICD) or ADAM17 effectively reverse the inhibitory effect of RECK in CD133‐positive cells. More importantly, RECK reduces tumorigenic activity of CD133‐positive cells in vivo. Conversely, knockdown of RECK in non‐tumorigenic GI2 cells increases stemness and CD133 expression and sphere forming ability. Collectively, these results indicate that RECK represses stemness gene expression and stem‐like properties by inhibiting ADAM‐mediated Notch1 shedding and activation. J. Cell. Physiol. 229: 191–201, 2014. © 2013 Wiley Periodicals, Inc.
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