【摘要】 目的:探讨细胞因子CC配体2(CCL-2)对急性白血病细胞THP-1迁移和侵袭的影响及机制。方法:采用慢病毒包装的方法将表达质粒PLVX-CCL2转染THP-1细胞,用RT-PCR检测CCL-2 RNA水平的表达,Western blot及ELISA法验证蛋白水平的变化,应用Transwell迁移和侵袭实验方法分别检测CCL2重组蛋白及THP-1自身高表达CCL2对THP-1细胞迁移和侵袭能力的影响,应用迁移相关细胞因子的PCR array法探讨可能的机制。结果:RT-PCR和Western blot方法证实成功构建过表达CCL2的THP-1-CCL2细胞。ELISA法检测显示,THP-1-CCL2培养上清中CCL2蛋白水平显著增高;高表达CCL2的转染组细胞在Trans-Matrigel实验中,其上室CCL2蛋白浓度显著高于下室(P<0.001),同时穿膜能力下降,但迁移能力没有明显改变,与对照组比较,分别为(1.702±0.537 vs0.520±0.255)(P=0.026)和(9.293±0.302 vs 9.187±0.526)(P=0.77),增加Trans-Matrigel下室的CCL2浓度,可以增强THP-1细胞的迁移能力和侵袭能力。迁移RT2 profiler PCR array显示,CCL2重组蛋白处理THP-1细胞后,与对照组相比CCL2、EPX、SPP1、CX3CL1和CXCL13的表达上调(>2倍)。结论:CCL2通过白血病细胞THP-1的自分泌多种炎性趋化因子而调控其迁移和侵袭。更多还原

【Abstract】 Objective: To investigate the effects and mechanism of CCL2 on the migration and invasion of human leukemia cell THP-1. Methods: The CCL2 gene was recombined with the transfer plasmid-PLVX and transfected into THP-1 cells. The CCL2 expression at RNA level was detected by RT-PCR,the CCL2 expression at protein level was determined by Western blot and ELISA,the influence of overexpression of CCL2 recombinant protein and THP-1 cells on the migration and invasion ability of THP-1 cells was analyzed by transwell migration and invasion tests,the PCRarray of migration-related cytokines was used to clarify the patential mechanism. Results: With the Trans-Matrigel assay,the concentration of CCL2 in THP-1 transfected with CCL2 in the upper cells was higher than that in the lower cells,meanwhile,the invasion ability of CCL2-transfected THP-1 cells decreased. Increasing recombinant protein of CCL2( rpCCL2) in the lower cells promoted migration of THP-1 cells. Migration RT2 profiler PCR array showed that the cells treated with rpCCL2 had higher levels of expression of CCL2,EPX,SPP1,CX3 CL1 and CXCL13,as compared with control group. Conclusion: CCL2 affects the migration and invasion of THP-1 by autosecreting a series of inflammatory chemokines.更多还原