CAJ | 학술논문

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。
목적：탐토초고분자량취을희（ UHMWPE）과립대인단핵세포주THP-1、성섬유양활막세포（ FLS）공배양체계천이、침습능력급대세포외기질금속단백매유도제（ EMMPRIN）여기질금속단백매（ MMPs）표체적영향。방법건립THP-1-FLS세포공배양체계，응용태반람염색검측THP-1세포、FLS세포존활솔，Western blot、명효매보、실시정량PCR방법검측양품중적EMMPRIN、MMP-2、MMP-9적표체급활성，병응용RNA간우기술평고EMMPRIN적작용。결과 UHMWPE과립자격THP-1세포EMMPRIN표체상조，대FLS적EMMPRIN표체무명현영향。 UHMWPE과립자격FLS적MMP-2、MMP-9표체상조、활성증강，대 THP-1표체 MMPs 무명현영향。 THP-1-FLS 공배양시， UHMWPE과립자격사EMMPRIN、MMP-2、MMP-9표체상조경위현저。과립자격하，THP-1-FLS공배양체계적천이능력、침습능력명현강우무과립자격，병여EMMPRIN적표체수평화MMPs적활성정명현정상관。행THP-1세포EMMPRIN기인간우후，공배양체계적EMMPRIN、MMPs표체균하조，천이、침습능력하강。결론 UHMWPE 과립가자격THP-1-FLS 공배양체계고표체EMMPRIN，상조MMP-2、MMP-9적분비병증강기활성，제고공배양체계적천이、침습능력。
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 .