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| Wnt3诱导小鼠肝前体细胞上皮-间质转化 |
| 胡代曦1,2,张锡峰1,2,张超1,2,钟沁1,2,冯涛1,2,黄佳祎1,3* |
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(1. 重庆医科大学分子医学与肿瘤研究中心,重庆 400016
2. 重庆医科大学基础医学院生物化学与分子生物学教研室,重庆 400016
3. 重庆医科大学基础医学院病理生理学教研室,重庆 400016
*通信作者) |
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| 摘要: |
| 目的 探讨wnt3对小鼠肝前体细胞(14-19)发生上皮-间质转化的影响。方法 分别将空载体Ad-GFP和表达wnt3的腺病毒Ad-GFP-wnt3转入小鼠肝前体细胞中,显微镜下观察细胞形态变化。细胞划痕实验和细胞迁移实验观察14-19细胞迁移能力。实时荧光定量PCR和蛋白质印迹分析分别检测14-19细胞中上皮标记物和间质标记物的表达改变。结果 显微镜下可见高表达wnt3的14-19细胞由不规则多边形变为长梭形。细胞划痕实验和细胞迁移实验结果显示,高表达wnt3的14-19细胞迁移能力明显提高,与空载体转染细胞相比差异具有统计学意义(P<0.01)。 实时荧光定量PCR和蛋白质印迹结果显示,间质标记物N-cadherin、vimentin和twist1的mRNA和蛋白水平表达上调,相反上皮标记物E-cadherin的mRNA水平和蛋白水平表达下调, 与空载体转染细胞相比差异具有统计学意义(P<0.01)。结论 Wnt3能够促使小鼠肝前体细胞发生上皮-间质转化,提示其可能参与了肝纤维化的发展进程。 |
| 关键词: wnt3 肝前体细胞 上皮-间质转化 肝硬化 |
| DOI： |
| 投稿时间：2013-06-22修订日期：2013-07-15 |
| 基金项目:国家自然科学基金(81071770,81201679). |
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| Wnt3-induced epithelial-mesenchymal transition in mouse hepatic progenitor cells |
| HU Dai xi1,2,ZHANG Xi feng1,2,ZHANG Chao1,2,ZHONG Qin1,2,FENG Tao1,2,HUANG Jia yi1,3* |
(1. Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016, China
2. Department of Biochemistry and Molecular Biology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, China
3. Department of Pathophysiology, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, China
*Corresponding author.) |
| Abstract: |
| Objective To investigate the impact of wnt3 upon epithelial-mesenchymal transition in mouse hepatic progenitor cells(14-19 cells). Methods Ad-GFP-wnt3 virus and blank Ad-GFP were transfected into 14-19 cells; cell morphology was observed under microscope. The migration ability of 14-19 cells was examined by wound-healing assay and transwell assay. The expression of epithelial and mesenchymal markers in 14-19 cells was detected by real-time PCR and Western blotting analysis. Results Microscopic observation found that 14-19 cells changed from an epithelial characteristic shape into a spindle-like shape. Wound-healing assay and Transwell assay showed that the migration ability of 14-19 cells highly expressing wnt3 was significantly enhanced compared with the blank control group(P<0.01). Real-time PCR and Western blotting analysis showed that mesenchymal markers N-cadherin, vimentin and twist1 were significantly increased at both mRNA and protein levels in Ad-GFP-wnt3 virus group than the blank control group, while the epithelial marker E-cadherin was significantly decreased at mRNA and protein levels(P<0.01). Conclusion Wnt3 can induce epithelial-mesenchymal transition in mouse hepatic progenitor cells, indicating that wnt3 may take part in the progress of hepatic fibrosis. |
| Key words: wnt3 mouse hepatic progenitor cells epithelial-mesenchymal transition liver cirrhosis |
