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Periostin过表达对鼻咽癌6-10B细胞侵袭和迁移的影响及机制
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*
解剖学报 ›› 2014, Vol. 45 ›› Issue (4) : 500-506.
Periostin过表达对鼻咽癌6-10B细胞侵袭和迁移的影响及机制
Effects and mechanisms of periostin overexpression on invasion and migration of the nasopharyngeal carcinoma 6-10B cell line
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integrin αvβ5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，integrin αvβ5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，integrin αvβ5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integrin αvβ5受体结合而提高MMPs的活性。
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.
Periostin / 鼻咽癌 / 转移 / 转染 / Transwell / 免疫组织化学 / 人
Periostin / Nasopharyngeal cancer / Metastasis / transfection / Transwell / Immunohistochemistry / Human
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国家自然科学基金;Delta原钙黏蛋白（delta-protocadherin）在脊髓发育过程中的表达和功能调控研究;湖南省自然科学衡阳联合基金资助项目;湖南省科技厅项目;南华大学博士启动基金
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