放射线照射的肺成纤维细胞对人脐带间充质干细胞中经典 Wnt/β-catenin 通路的影响

目的:体外研究放射线照射后的人肺成纤维细胞( HLFs )对人脐带间充质干细胞(HUMSCs)中经典Wnt/β-catenin信号通路的影响。方法:实验细胞分为共培养组和单纯 HUMSCs 组,共培养组为5 Gy X 射线照射后的 HLFs 通过Transwell系统与HUMSCs共培养。细胞培养3d后,蛋白质印迹法检测HUMSCs中GSK-3β、p-GSK-3β、FRAT1和胞核内β-catenin蛋白表达,酶联免疫吸附试验检测细胞培养上清液中WISP-1蛋白表达。结果:共培养组HUMSCs 中p-GSK-3β/GSK-3β相对比值(0.15±0.05)和FRAT1表达水平(0.48±0.0...

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Bibliographic Details
Published in浙江大学学报(医学版) no. 2; pp. 162 - 166
Main Author 张春阳 祝艳 冯华松 陈旭昕
Format Journal Article
LanguageChinese
Published 海军总医院呼吸内科,北京100048%海军总医院呼吸内科,北京,100048 2015
解放军医学院研究生队,北京100853
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ISSN1008-9292
DOI10.3785/j.issn.1008-9292.2015.03.007

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Summary:目的:体外研究放射线照射后的人肺成纤维细胞( HLFs )对人脐带间充质干细胞(HUMSCs)中经典Wnt/β-catenin信号通路的影响。方法:实验细胞分为共培养组和单纯 HUMSCs 组,共培养组为5 Gy X 射线照射后的 HLFs 通过Transwell系统与HUMSCs共培养。细胞培养3d后,蛋白质印迹法检测HUMSCs中GSK-3β、p-GSK-3β、FRAT1和胞核内β-catenin蛋白表达,酶联免疫吸附试验检测细胞培养上清液中WISP-1蛋白表达。结果:共培养组HUMSCs 中p-GSK-3β/GSK-3β相对比值(0.15±0.05)和FRAT1表达水平(0.48±0.07)较单纯HUMSCs组(0.55±0.05和1.16±0.13)明显下降,共培养组 HUMSCs 胞核内β-catenin (0.50±0.07)也较单纯HUMSCs组(2.39±0.15)明显下降;共培养组HUMSCs培养上清液中 WISP-1蛋白表达[(602.23±161.47) ng/mL ]较单纯 HUMSCs 组[(977.77±110.56)ng/mL]明显下降,差异均有统计学意义(均P<0.05)。结论:放射线照射后的HLFs对HUMSCs中经典Wnt/β-catenin通路具有抑制作用。
Bibliography:33-1248/R
Objective:To investigate the effect of irradiated human lung fibroblasts ( HLFs) on the canonical Wnt/β-catenin signaling pathway in human umbilical cord mesenchymal stem cells (HUMSCs).Methods:HUMSCs were cultured alone (single group) or co-cultured with HLFs exposed to 5Gy X-rays ( co-culture group ).The protein levels of GSK-3β, p-GSK-3β, FRAT1 and β-catenin in HUMSCs were examined by Western blotting 3 days after culture or co-culture.WISP-1 protein levels in conditioned medium were examined by ELISA .Results: The levels of p-GSK3β/GSK3β(0.15 ±0.05), FRAT1(0.48 ±0.07) and β-catenin(0.50 ±0.07) in co-cultured HUMSCs significantly decreased compared to those in single group ( 0 .55 ± 0.05, 1.16 ±0.13 and 2.39 ±0.15, all P〈0.05).The supernatant level of WISP-1 in co-culture group was significantly decreased [(602.23 ±161.47)ng/mL], compared to the single group [(977.77 ±110.56) ng/mL, P〈0.05].Conclusion: Irradiated HLFs attenuate the activation of canonical Wnt/β-catenin signaling pathway i
ISSN:1008-9292
DOI:10.3785/j.issn.1008-9292.2015.03.007