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。

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＆lt;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 .