| Periostin过表达对鼻咽癌6-10B细胞侵袭和迁移的影响及机制 |
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| 引用本文： | 王惠洁,石金凤,谢远杰,曾谷清,杜雅兰,黄欣琼,龙治峰,庾江东,李美香.Periostin过表达对鼻咽癌6-10B细胞侵袭和迁移的影响及机制[J].解剖学报,2014,0(4):500-506. |
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| 作者姓名： | 王惠洁 石金凤 谢远杰 曾谷清 杜雅兰 黄欣琼 龙治峰 庾江东 李美香 |
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| 作者单位： | 1. 南华大学医学院组织学胚胎学教研室；2. 细胞分化和组织发生研究室，湖南 衡阳 421001;3. 南华大学护理学院护理研究所，湖南 衡阳 421001; 4. 衡阳市中心医院耳鼻喉科,湖南 衡阳 421001 |
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| 基金项目： | 国家自然科学基金资助项目(81072199，81272959)；湖南省自然科学衡阳联合基金资助项目(12JJ9034)；湖南省科技厅资助项目(2011SK3088)；湖南省大学生创新课题资助项目(项目编号：2012cshncx1) |
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| 摘 要： | 目的探讨外源表达periostin蛋白对鼻咽癌(NPC)6-10B细胞侵袭和迁移能力的影响及机制。方法采用脂质体转染技术建立稳定高表达periostin的6-10B细胞,RT-PCR结合Western blotting检测转染效率,Tanswell分析periostin高表达后6-10B细胞侵袭和迁移能力的改变,明胶酶谱法检测基质金属蛋白酶(MMPs)活性的变化;免疫组织化学检测整合素(integrin)αvβ5在转染periostin前后6-10B细胞中的表达以及其与periostin在鼻咽癌和正常鼻咽黏膜组织中的表达,image-pro Plus 6.0软件进行图像分析及吸光度测定,统计学分析鼻咽癌组织中periostin和integrinαvβ5表达强度的相关性。结果 Periostin过表达的6-10B细胞侵袭、迁移能力明显增强,细胞培养上清中MMP-2、MMP-9活性增加,integrinαvβ5在periostin过表达的6-10B细胞及NPC组织的表达明显强于对照组,Spearman相关性分析显示,integrinαvβ5和periostin在NPC组织中的表达强度成正相关(r=0.682,P=0.000)。结论外源表达periostin能促进NPC的侵袭和转移,其机制可能是通过与NPC细胞膜上integrinαvβ5受体结合而提高MMPs的活性。
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| 关 键 词： | Periostin 鼻咽癌 转移 转染 Transwell 免疫组织化学 人 |
| 收稿时间： | 2013-09-24 |
Effects and mechanisms of periostin overexpression on invasion and migration of the nasopharyngeal carcinoma 6-10B cell line |
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| WANG Hui-jie SHI Jin-feng XIE Yuan-jie,ZENG Gu-qing DU Ya-lan HUANG Xing-qiong LONG Zhi-feng YU Jiang-dong LI Mei-xiang.Effects and mechanisms of periostin overexpression on invasion and migration of the nasopharyngeal carcinoma 6-10B cell line[J].Acta Anatomica Sinica,2014,0(4):500-506. |
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| Authors: | WANG Hui-jie SHI Jin-feng XIE Yuan-jie ZENG Gu-qing DU Ya-lan HUANG Xing-qiong LONG Zhi-feng YU Jiang-dong LI Mei-xiang |
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| Institution: | 1. Department of Histology and Embryology, School of Medicine, University of South China; 2. Laboratory of Cell Differentiation and Apoptosis, School of Medicine, University of South China, Hu’nan Hengyang 421001, China;3. Nursing Research Institute，School of Life Science and Technology, University of South China, Hu’nan Hengyang 421001, China;4. Otolaryngology Department of Hengyang Central Hospital； Hu’nan Hengyang 421001, China;
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| Abstract: | Objective To explore the effects and mechanisms of periostin overexpression on migration and invasion of nasopharyngeal carcinoma(NPC) cell line. Methods The recombinant plasmids ［pCMV-neo (+)-periostin］ and control plasmids ［pCMV-neo (+)］ were transfected into 6-10B cells using lipofectamine 2000TM reagent. The expression of periostin was detected with PCR and Western blotting. Transwell chamber invasion assay was employed to assay the migration and invasion of 6-10B cells before and after transfection. A gelatin zymogram was used to detect the activity of MMP-2 and MMP-9 in cultivated supernatant of 6-10B cells before and after transfection. The expression of integrin-αvβ5 was detected by immunohistochemistry(IHC) in 6-10Bperiostin cells, 6-10Bvector cells and 6-10B cells as well as normal nasopharyngeal mucosa (NNM) and NPC and at the same time periostin also was detected by immumohistochemistry in NNM and NPC, and densitometry analysis using image-pro plus 6.0 software, and the correlation between periostin and integrin-αvβ5 on NPC was assayed with statistics. Results Over expression of periostin promoted cell migration and invasion. The expression levels of integrin-αvβ5 in primary NPC and 6-10Bperiostin cells were significantly higher than those in NNM and 6-10Bvector, 6-10B cells. The expression in NPC of integrin-αvβ5 showed positively correlated with the expression of periostin (r=0.682, P<0.01). Conclusion Periostin plays an important role in regulation of cell migration and invasion probably by combining with integrin-αvβ5 to improve the activities of MMPs. |
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| Keywords: | Periostin Nasopharyngeal cancer Metastasis transfection Transwell Immunohistochemistry Human |
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