CAJ | 학술논문

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，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integrinαvβ5受体结合而提高MMPs的活性。
목적：탐토외원표체periostin단백대비인암（NPC）6-10B세포침습화천이능력적영향급궤제。방법채용지질체전염기술건립은정고표체periostin적6-10B세포，RT-PCR결합Western blotting검측전염효솔，Tanswell분석periostin고표체후6-10B세포침습화천이능력적개변，명효매보법검측기질금속단백매（ MMPs）활성적변화；면역조직화학검측정합소（integrin）αvβ5재전염periostin전후6-10B세포중적표체이급기여periostin재비인암화정상비인점막조직중적표체， image-pro Plus 6．0연건진행도상분석급흡광도측정，통계학분석비인암조직중periostin화integrinαvβ5표체강도적상관성。결과 Periostin과표체적6-10B세포침습、천이능력명현증강，세포배양상청중MMP-2、MMP-9활성증가，integrin αvβ5재periostin과표체적6-10B세포급NPC조직적표체명현강우대조조，Spearman상관성분석현시， integrin αvβ5화 periostin 재 NPC 조직중적표체강도성정상관（ r ＝0．682， P ＝0．000）。결론외원표체periostin능촉진NPC적침습화전이，기궤제가능시통과여NPC세포막상integrinαvβ5수체결합이제고MMPs적활성。
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 .