Refractoriness of interferon-beta signaling through NOD1 pathway in mouse respiratory epithelial cells using the anticancer xanthone compound  

作　　者：Zaifang Yu Jarrod D Predina Guanjun Cheng 

机构地区：[1]Thoracic Oncology Research Laboratory,1016B ARC,University of Pennsylvania [2]Department of Surgery,Thoracic Surgery Research Laboratory,University of Pennsylvania

出　　处：《World Journal of Biological Chemistry》2013年第2期18-29,共12页世界生物化学杂志（英文版）（电子版）

基　　金：Supported by NCI PO1 CA 66726

摘　　要：AIM:To explore the possibility that nucleotide oligomerization domain 1(NOD1) pathway involved in refractoriness of interferon-β signaling in mouse respiratory epithelial cells induced by the anticancer xanthone compound,5,6-dimethylxanthenone-4-acetic acid(DMXAA).METHODS:C10 mouse bronchial epithelial cells were grown in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum,2 mmol/L glutamine,100 units/mL penicillin,100 g/mL streptomycin.Pathogen-free female BALB/c mice were used to explore the mechanisms of refractoriness of interferon-signaling.Mouse thioglycollate-elicited peritoneal macrophages,bone marrow derived macrophages and bone marrow derived dendritic cells were collected and cultured.The amount of interferon(IFN)-inducible protein-10(IP10/CXCL10),macrophage chemotactic protein(MCP1/CCL2) and interleukin(IL)-6 secreted by cells activated by DMXAA was quantified using enzyme-linked immunosorbent assay kits according to the instructions of the manufacturers.Total RNA was isolated from cells or nasal epithelium with RNeasy Plus Mini Kit,and cDNA was synthesized.Gene expression was checked using Applied Biosystems StepOne Real-Time Polymerase Chain Reaction System.Transfection of small interfering RNA(siRNA) control,NOD1 duplexed RNA oligonucleotides,and high-mobility group box 1/2/3(HMGB1/2/3) siRNA was performed using siRNA transfection reagent.RESULTS:DMXAA activates IFN-β pathway with high level of IFN-β dependent antiviral genes including 2',5'-oligoadenylate synthetase 1 and myxovirus resistance 1 in mouse thioglycollate-elicited peritoneal macrophages,bone marrow derived macrophages and bone marrow derived dendritic cells.Activation of IFN-β by DMXAA involved in NOD1,but not HMGB1/2/3 signal pathway demonstrated by siRNA.NOD1 pathway plays an important role in refractoriness of IFN-β signaling induced by DMXAA in mouse C10 respiratory epithelial cells and BALB/c mice nasal epithelia.These data indicate that DMXAA is not well adapted to the intrinsic properties of IFN-β AIM:To explore the possibility that nucleotide oligomerization domain 1(NOD1) pathway involved in refractoriness of interferon-β signaling in mouse respiratory epithelial cells induced by the anticancer xanthone compound,5,6-dimethylxanthenone-4-acetic acid(DMXAA).METHODS:C10 mouse bronchial epithelial cells were grown in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum,2 mmol/L glutamine,100 units/mL penicillin,100 g/mL streptomycin.Pathogen-free female BALB/c mice were used to explore the mechanisms of refractoriness of interferon-signaling.Mouse thioglycollate-elicited peritoneal macrophages,bone marrow derived macrophages and bone marrow derived dendritic cells were collected and cultured.The amount of interferon(IFN)-inducible protein-10(IP10/CXCL10),macrophage chemotactic protein(MCP1/CCL2) and interleukin(IL)-6 secreted by cells activated by DMXAA was quantified using enzyme-linked immunosorbent assay kits according to the instructions of the manufacturers.Total RNA was isolated from cells or nasal epithelium with RNeasy Plus Mini Kit,and cDNA was synthesized.Gene expression was checked using Applied Biosystems StepOne Real-Time Polymerase Chain Reaction System.Transfection of small interfering RNA(siRNA) control,NOD1 duplexed RNA oligonucleotides,and high-mobility group box 1/2/3(HMGB1/2/3) siRNA was performed using siRNA transfection reagent.RESULTS:DMXAA activates IFN-β pathway with high level of IFN-β dependent antiviral genes including 2’,5’-oligoadenylate synthetase 1 and myxovirus resistance 1 in mouse thioglycollate-elicited peritoneal macrophages,bone marrow derived macrophages and bone marrow derived dendritic cells.Activation of IFN-β by DMXAA involved in NOD1,but not HMGB1/2/3 signal pathway demonstrated by siRNA.NOD1 pathway plays an important role in refractoriness of IFN-β signaling induced by DMXAA in mouse C10 respiratory epithelial cells and BALB/c mice nasal epithelia.These data indicate that DMXAA is not well adapted to the intrinsic properties of

关 键 词：INNATE immunity Interferon REFRACTORINESS XANTHONE BRONCHIAL EPITHELIUM 

分 类 号：R73-3[医药卫生—肿瘤]