巨噬细胞来源的外泌体对弥漫大B细胞淋巴瘤增殖、迁移、侵袭和凋亡的影响

Effect of macrophage-derived exosomes on proliferation,migration,invasion,and apoptosis of diffuse large B-cell lymphoma

目的探讨巨噬细胞来源的外泌体对弥漫大B细胞淋巴瘤(DLBCL)增殖、迁移、侵袭和凋亡的影响。方法构建体外巨噬细胞极化模型;提取外泌体,以加外泌体的DLBCL细胞作为外泌体组,以未加外泌体的DLBCL细胞作为对照组;采用透射电镜检测外泌体形态;采用纳米颗粒跟踪分析技术检测外泌体粒径与水平;采用免疫荧光法检测外泌体传递;采用CCK-8法检测人DLBCL细胞系U2932细胞增殖;采用细胞划痕试验检测U2932细胞迁移;采用Transwell法检测U2932细胞侵袭;采用流式细胞术检测U2932细胞凋亡。结果外泌体水平为(5.27±0.36)×10^(10)particles/mL,平均粒径为121.69 nm,单峰,呈正态分布,直径50~150 nm,其密度约为(1.15±0.03)g/mL。与对照组比较,外泌体组在人DLBCL细胞和巨噬细胞株RAW264.7细胞中的传递均明显增多,差异均有统计学意义(P<0.05);外泌体组人DLBCL细胞系U2932细胞增殖、迁移和侵袭能力均明显高于对照组,差异均有统计学意义(P<0.05);外泌体组人DLBCL细胞系U2932细胞凋亡能力明显低于对照组,差异有统计学意义(P<0.05)。结论巨噬细胞来源的外泌体可以促进人DLBCL细胞系U2932细胞的增殖、迁移和侵袭,并抑制其凋亡,为进一步研究巨噬细胞来源外泌体在人DLBCL发生和发展中的相关机制提供了一定的理论基础。

Objective To investigate the effects of macrophage-derived exosomes on proliferation,migration,invasion and apoptosis of diffuse large B-cell lymphoma(DLBCL).Methods An in vitro macrophage polarization model was established.Exosomes were extracted,and DLBCL cells with exosomes were selected as exosome group,LBCL cells without exosomes were selected as control group.Transmission electron microscopy was used to detect the morphology of exosomes.Nanoparticle tracking analysis was used to detect the particle size and levels of exosomes.The proliferation of human DLBCL cell line U2932 was detected by CCK-8 method.DLBCL cells with exosomes were used as exosome group,and DLBCL cells without exosomes were used as control group.The migration of U2932 cells was detected by cell scratch test.Transwell assay was used to detect the invasion of U2932 cells.The apoptosis of U2932 cells was detected by flow cytometry.Results The levels of exosomes was(5.27±0.36)×10^(10) particles/mL,the average size was 121.69 nm,the diameter was 50-150 nm,and the density was(1.15±0.03)g/mL.Compared with control group,the transmission of exosomes in human DLBCL cells and macrophage cell line RAW264.7 cells was significantly increased,and the differences were statistically significant(P<0.05).The proliferation,migration and invasion abilities of human DLBCL cell line U2932 cells in exosome gr oup were significantly higher than those in control group(P<0.05).The apoptosis ability of human DLBCL cell line U2932 in exosome group was significantly lower than that in control group,and the difference was statistically significant(P<0.05).Conclusion Macrophage-derived exosomes can promote the proliferation,migration and invasion of human DLBCL cell line U2932,and inhibit its apoptosis,which provides a theoretical basis for further study on the mechanism of macrophage-derived exosomes in the occurrence and development of human DLBCL.