Angiopoietin-like protein 1 suppresses SLUG to inhibit cancer cell motility
Tsang-Chih Kuo, … , Jen-Liang Su, Min-Liang Kuo
Tsang-Chih Kuo, … , Jen-Liang Su, Min-Liang Kuo
Published February 22, 2013
Citation Information: J Clin Invest. 2013;123(3):1082-1095. https://doi.org/10.1172/JCI64044.
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Research Article

Angiopoietin-like protein 1 suppresses SLUG to inhibit cancer cell motility

Abstract

Angiopoietin-like protein 1 (ANGPTL1) is a potent regulator of angiogenesis. Growing evidence suggests that ANGPTL family proteins not only target endothelial cells but also affect tumor cell behavior. In a screen of 102 patients with lung cancer, we found that ANGPTL1 expression was inversely correlated with invasion, lymph node metastasis, and poor clinical outcomes. ANGPTL1 suppressed the migratory, invasive, and metastatic capabilities of lung and breast cancer cell lines in vitro and reduced metastasis in mice injected with cancer cell lines overexpressing ANGPTL1. Ectopic expression of ANGPTL1 suppressed the epithelial-to-mesenchymal transition (EMT) by reducing the expression of the zinc-finger protein SLUG. A microRNA screen revealed that ANGPTL1 suppressed SLUG by inducing expression of miR-630 in an integrin α1β1/FAK/ERK/SP1 pathway–dependent manner. These results demonstrate that ANGPTL1 represses lung cancer cell motility by abrogating the expression of the EMT mediator SLUG.

Authors

Tsang-Chih Kuo, Ching-Ting Tan, Yi-Wen Chang, Chih-Chen Hong, Wei-Jiunn Lee, Min-Wei Chen, Yung-Ming Jeng, Jean Chiou, Pei Yu, Pai-Sheng Chen, Ming-Yang Wang, Michael Hsiao, Jen-Liang Su, Min-Liang Kuo

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Figure 7

ANGPTL1 induces miR-630 transcription through repression of the ERK/SP1 pathway.

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ANGPTL1 induces miR-630 transcription through repression of the ERK/SP1 ...
(A) qRT-PCR analysis of miR-630 expression in ANGPTL1-overexpressing CL1-5 cells transiently transfected with myr-AKT and MEK1. (B) qRT-PCR analysis of pre–miR-630 and pri–miR-630 expression in ANGPTL1-overexpressing CL1-5 cells transiently transfected with myr-AKT and MEK1. (C) Schematic representation of various miR-630 promoter reporters (left). CL1-5/pcDNA3.1 and CL1-5/ANGPTL1 cells were transfected with various miR-630 promoter reporters or the pGL3-basic vector, and the luciferase activity was measured (right). (D) Schematic representation of the 46-bp region of human miR-630 promoter (–137 to –92 bp), where underlining indicates potential binding sites (top). Quantitative ChIP (qChIP) analysis of the miR-630 promoter region in ANGPTL1-overexpressing CL1-5 cells transiently transfected with myr-AKT and MEK1 (bottom). Signal is relative to the CL1-5/pcDNA3.1 plus vector group. (E) Schematic representation of the 46-bp region of human miR-630 promoter (–137 to –92 bp), where underlining indicates the SP1-binding site and mutated SP1-binding site (top). CL1-5/pcDNA3.1 and CL1-5/ANGPTL1 cells were transfected with miR-630 promoter reporter (–137 to –92 bp), miR-630 promoter reporter (–137 to –92 bp) SP1 mutant construct (MT SP1), or the pGL3-basic vector, and the luciferase activity was measured (bottom). (F) Western blot analysis of SP1 and SLUG, and qRT-PCR analysis of miR-630 expression in ANGPTL1-overexpressing CL1-5 cells infected with shRNA-Luc and shRNA-SP1. Data are shown as mean ± SEM. *P < 0.05, **P < 0.01 (2-tailed Student’s t test).