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Supplementary Methods from ALX1 Induces Snail Expression to Promote Epithelial-to-Mesenchymal Transition and Invasion of Ovarian Cancer Cells

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posted on 2023-03-30, 22:00 authored by Hong Yuan, Hiroaki Kajiyama, Satoko Ito, Nobuhisa Yoshikawa, Toshinori Hyodo, Eri Asano, Hitoki Hasegawa, Masao Maeda, Kiyosumi Shibata, Michinari Hamaguchi, Fumitaka Kikkawa, Takeshi Senga

PDF file - 46K, Includes information regarding plasmids, In situ hybridization, Immunofluorescence and immunoblot analysis, as well as RT-PCR

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ARTICLE ABSTRACT

Ovarian cancer is a highly invasive and metastatic disease with a poor prognosis if diagnosed at an advanced stage, which is often the case. Recent studies argue that ovarian cancer cells that have undergone epithelial-to-mesenchymal transition (EMT) acquire aggressive malignant properties, but the relevant molecular mechanisms in this setting are not well-understood. Here, we report findings from an siRNA screen that identified the homeobox transcription factor ALX1 as a novel regulator of EMT. RNA interference–mediated attenuation of ALX1 expression restored E-cadherin expression and cell–cell junction formation in ovarian cancer cells, suppressing cell invasion, anchorage-independent growth, and tumor formation. Conversely, enforced expression of ALX1 in ovarian cancer cells or nontumorigenic epithelial cells induced EMT. We found that ALX1 upregulated expression of the key EMT regulator Snail (SNAI1) and that it mediated EMT activation and cell invasion by ALX1. Our results define the ALX1/Snail axis as a novel EMT pathway that mediates cancer invasion. Cancer Res; 73(5); 1581–90. ©2012 AACR.