UHMWPE 颗粒上调 EMMPRIN、MMPs 表达并增强共培养体系中 THP-1、FLS 迁移、侵袭能力的实验研究

目的：探讨超高分子量聚乙烯（ UHMWPE）颗粒对人单核细胞株THP-1、成纤维样滑膜细胞（ FLS）共培养体系迁移、侵袭能力及对细胞外基质金属蛋白酶诱导剂（ EMMPRIN）与基质金属蛋白酶（ MMPs）表达的影响。方法建立THP-1-FLS细胞共培养体系，应用台盼蓝染色检测THP-1细胞、FLS细胞存活率，Western blot、明胶酶谱、实时定量PCR方法检测样品中的EMMPRIN、MMP-2、MMP-9的表达及活性，并应用RNA干扰技术评估EMMPRIN的作用。结果 UHMWPE颗粒刺激THP-1细胞EMMPRIN表达上调，对FLS的EMMPRIN表达无明显影响。 UHMWPE颗粒刺激FLS的MMP-2、MMP-9表达上调、活性增强，对 THP-1表达 MMPs 无明显影响。 THP-1-FLS 共培养时， UHMWPE颗粒刺激使EMMPRIN、MMP-2、MMP-9表达上调更为显著。颗粒刺激下，THP-1-FLS共培养体系的迁移能力、侵袭能力明显强于无颗粒刺激，并与EMMPRIN的表达水平和MMPs的活性呈明显正相关。行THP-1细胞EMMPRIN基因干扰后，共培养体系的EMMPRIN、MMPs表达均下调，迁移、侵袭能力下降。结论 UHMWPE 颗粒可刺激THP-1-FLS 共培养体系高表达EMMPRIN，上调MMP-2、MMP-9的分泌并增强其活性，提高共培养体系的迁移、侵袭能力。

Ultra-high molecular weight polythylene particles promote migration and invasion ability of co- cultured TI-IP-1 cells and fibroblast-like synoviocytes by upregulating expression of extracellular matrix metalloproteinase inducer and matrix metalloproteinases

Objective To investigate the effects of ultra-high molecular weight polythylene （ UHMWPE） particles on the migration , the invasion ability and the expression of extracellular matrix metalloproteinase inducer （ EMMPRIN）, matrix metalloproteinase （ MMP）-2, and MMP-9 of the co-cultured THP-1 cells and fibroblast-like synoviocytes （ FLS） in vitro.Methods A co-cultured system of THP-1-FLS was established .The migration and invasion abilities of the cells were tested by transwell assays.Western blot analysis and zymographic analysis were adopted to detect EMMPRIN and MMPs .Real time PCR （RT-PCR） was used to determine the level of mRNAs of EMMPRIN , MMP-2, and MMP-9.The RNA interference was applied to evaluate the function of EMMPRIN .Results UHMWPE particles upregulated EMMPRIN expression in THP-1 cells but not FLS .UHMWPE particles upregulated MMP-2 and MMP-9 expression , also enhanced their activities in FLS while THP-1 cells were not involved .In the circumstance of co-culture, the upregulations mentioned above mediated by the particles were much more significant .In the presence of UHMWPE particles , the migration and invasion abilities of the cells increased significantly in the THP-1-FLS co-cultured system compared with that in a particle-absent co-culture system . The migration and invasion abilities were found to be positively-correlated with the EMMPRIN expression and MMPs activities .After RNA interference targeting EMMPRIN gene in THP-1 cells, the expression of EMMPRIN and MMPs as well as the migration and invasion abilities of the co-culture system were attenuated distinctly .Conclusion UHMWPE particles promote the migration and invasion abilities of the co-cultured THP-1 cells and FLS through an enhanced expression of MMP-2, MMP-9 caused by the particle-upregulated EMMPRIN .