The activation of volume-activated chloride Cl- channels has been implicated to play important roles in modulating cell cycle and cell migration. The aim of this study was to determine whether volume-activated Cl- channels are involved in cell-cycle-dependent regulation of cell migration in HeLa cells. Using techniques including cell-cycle synchronization, transwell migration assays and the patch-clamp technique, we demonstrate in this study that both the expression of volume-activated chloride cur-rent (I-Cl,I-vol) and the potential of cell migration are cell-cycle-dependent; specifically, these events were high in G(0)/G(1) phase, low in S phase, and medium in G(2)/M phase. Moreover, the mean density of I-Cl,I-vol was positively correlated to the rate of cell migration during cell-cycle progression. Additionally, endogenous suppression of I-Cl,I-vol by transfecting cells with CIC-3 antisense oligonucleotides arrested cells in S phase and slowed cell migration. Collectively, our results suggest that volume-activated Cl- channels contribute to the cell-cycle-dependent regulation of cell migration. (C) 2008 Elsevier Inc. All rights reserved.
OBJECTIVE: An aneurysm is an inflammatory vascular condition. Phosphatidylinositol 3-kinases delta is highly expressed in leukocytes, and play a key role in innate immunity. However, the link between phosphatidylinositol 3-kinases delta and aneurysm development has not yet been elucidated. APPROACH AND RESULTS: Carotid ligation unexpectedly induced characteristic aneurysm formation beneath the ligation point in p110delta(D910A/D910A) mice (n=25; P<0.001 versus wild-type). Besides, p110delta inactivation exacerbated CaCl2-induced abdominal aortic aneurysms development. A reverse transcription polymerase chain reaction microarray revealed significant extracellular matrix components degradation and matrix metalloproteinases (MMPs) upregulation in the abdominal aorta of p110delta(D910A/D910A) mice. Similarly, the expression of both collagen I and IV was significantly decreased (n=10; P<0.05 versus wild-type) in carotid artery. Western blot assay confirmed that MMP-12 was significantly upregulated in arteries of p110delta(D910A/D910A) mice (n=10; P<0.01 versus wild-type). In vitro, p110delta inactivation marked increase peritoneal macrophages recruitment and synergistically enhance tumor necrosis factor-alpha-induced recruitment. A specific phosphatidylinositol 3-kinases delta inhibitor (IC87114) or genetic p110delta inactivation upregulated MMP-12 expression and c-Jun phosphorylation (n=6; P<0.05 versus wild-type macrophages). IC87114 also increased activator protein-1 DNA-binding activity (n=6; P<0.001 versus control) and enhanced the effect of tumor necrosis factor-alpha on activator protein-1-binding activity (n=5; P<0.01 versus tumor necrosis factor-alpha treatment groups). Knockdown of c-Jun suppressed the effect of the IC87114 and tumor necrosis factor-alpha on MMP-12 mRNA expression (n=5 in each group; P<0.01 versus scrRNA treatment groups). CONCLUSIONS: Our findings demonstrate that p110delta inactivation leads to extracellular matrix degradation in vessels and promotes aneurysm development by inducing macrophages migration and upregulating the activator protein-1/MMP-12 pathway in macrophages.
Journal of Biological Chemistry,2009年284(7):4473-4483 ISSN：0021-9258
[Slungaard, Arne; Gupta, Kalpna; Geng, Jian-Guo] Vascular Biology Center, Division of Hematology, Oncology and Transplantation, University of Minnesota Medical School, 420 Delaware St. S.E., Minneapolis, Minnesota 55455, United States;[Li, Yi-Dan; Zheng, Sheng-Xi; Wang, Jin-Tao; Chen, Ming; Ye, Bu-Qing; Liu, Ji-Guo; Geng, Jian-Guo] Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Shanghai 200031, China;[Wang, Li-Jing] Institute of Basic Medical Sciences, Guangdong Pharmaceutical University, Guangzhou, Guangdong 510006, China;[Wang, Jian-Guo; Mackman, Nigel; Key, Nigel S.; Pei, Xin-Hui] Division of Hematology/Oncology, University of North Carolina, Chapel Hill, NC 27599, United States;[Lin, Zhi-Lin; Pei, Xin-Hui] College of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China
Andrographolide - Chromatin immunoprecipitation - Deep vein thrombosis - Direct interactions - Heterodimer - In-vivo - Specific inhibitors - Tissue factors - Venous thrombosis
The Journal of biological chemistry,2009年284(7):4473-4483 ISSN：0021-9258
[Lin, Zhi-Xin; Pei, Xin-Hui] Shanghai Jiao Tong Univ, Coll Life Sci & Biotechnol, Shanghai 200240, Peoples R China.;[Geng, Jian-Guo; Wang, Jin-Tao; Ye, Bu-Qing; Zheng, Sheng-Xi; Chen, Ming; Li, Yi-Dan; Liu, Ji-Guo; Pei, Xin-Hui] Chinese Acad Sci, Grad Sch, Mol Cell Biol Lab, Inst Biochem & Cell Biol,Shangai Inst Biol Sci, Shanghai 200031, Peoples R China.;[Geng, Jian-Guo; Gupta, Kalpna; Slungaard, Arne] Univ Minnesota, Sch Med, Vasc Biol Ctr, Minneapolis, MN 55455 USA.;[Geng, Jian-Guo; Gupta, Kalpna; Slungaard, Arne] Univ Minnesota, Sch Med, Div Hematol Oncol & Transplantat, Minneapolis, MN 55455 USA.;[Geng, Jian-Guo] MMC 480,420 Delaware St SE, Minneapolis, MN 55455 USA.
[Geng, Jian-Guo] MMC 480,420 Delaware St SE, Minneapolis, MN 55455 USA.
NF-kappa B transcription factors regulate the expression of tissue factor (TF), a principal initiator of the coagulation cascade. Dominant among them is the p50/p65 heterodimer. Here we report that Andrographolide (Andro; a p50 inhibitor) and genetic deletion of p50 attenuated TF activity in stimulated endothelial cells and monocytes/macrophages. Results of the electrophoretic mobility "supershift" assay and chromatin immunoprecipitation demonstrated the direct interaction of the p50/p65 heterodimer with the NF-kappa B site of the human TF promoter. Andro-treated and p50 null mice both exhibited blunted TF expression and reduced venous thrombosis, which were recapitulated by an anti-murine TF antibody in vivo. Our findings thus indicate that regulation of TF by NF-kappa B transcription factor p50 is essential for the pathogenesis of deep vein thrombosis and suggest that specific inhibitors of p50, such as Andro, may be therapeutically valuable for preventing and perhaps treating venous thrombosis.