摘要
目的:探究结直肠癌来源的外泌体对巨噬细胞极化的影响及其作用机制。方法:建立THP-1来源的M2型巨噬细胞体外诱导分化模型,利用流式细胞染色实验检测细胞分化效率;通过与结直肠癌细胞共培养、结直肠癌细胞外泌体干预M2型巨噬细胞分化,利用实时荧光定量PCR实验检测ARG1、IL-10和NLRP3 mRNA的表达水平。结果:体外诱导分化的M2型THP-1巨噬细胞CD163阳性细胞比例显著升高;与对照组相比,与结直肠癌细胞HCT8和HCT116共培养的M2型巨噬细胞ARG1、IL-10表达水平显著升高,NLRP3表达水平下降。与转染阴性对照microRNA(miR-NC)组相比,转染miR-34a inhibitor的HCT8和HCT116共培养或外泌体刺激下,M2型巨噬细胞ARG1、IL-10表达水平下降,NLRP3表达水平升高;与miR-NC组相比,转染miR-34a inhibitor的THP-1细胞ARG1、IL-10表达水平显著下降,NLRP3表达水平显著升高。结论:结直肠癌细胞来源的外泌体miR-34a可以抑制巨噬细胞NLRP3的表达,促进巨噬细胞M2型极化。
Objective:To investigate the effect of colorectal cancer derived exosomes on macrophage polarization and the underlying mechanism.Methods:The THP-1-derived M2 macrophage differentiation model was established in vitro,and the cell differentiation efficiency was detected by flow cytometry assay.Colorectal cancer cells were co-cultured and colorectal cancer exosomes were isolated to intervene the differentiation of M2-type macrophages.The mRNA expression levels of ARG1,IL-10 and NLRP3 were detected by real-time qPCR.Results:The population of CD163+differentiated THP-1 cells was significantly increased.Compared with control group,the expression levels of ARG1 and IL-10 in M2 macrophages co-cultured with colorectal cancer cells HCT8 and HCT116 were significantly increased,while the expression level of NLRP3 was decreased.Compared with miR-NC group,the expression levels of ARG1 and IL-10 in M2 macrophages under the co-culture of HCT8 and HCT116 transfected with miR-34a inhibitor or the stimulation of exosomes were decreased,while the expression level of NLRP3 was increased.Compared with miR-NC group,the expression levels of ARG1 and IL-10 in THP-1 cells transfected with miR-34a inhibitor were significantly decreased,while the expression level of NLRP3 was significantly increased.Conclusion:The colorectal cancer cells derived-exosomal miR-34a could inhibit the expression of NLRP3 in macrophages and promote M2 macrophages polarization.
作者
倪红飞
李顾楠
张伟兴
李紫燕
NI Hongfei;LI Gunan;ZHANG Weixing;LI Ziyan(Department of Gastrointestinal Surgery,Changzhou Cancer Hospital,Changzhou 213000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2022年第24期2976-2979,共4页
Chinese Journal of Immunology
