外泌体介导miR-3473b靶向NF-κB对肺癌细胞肺内转移及B细胞数量的影响  

Effect of Exosomes-mediated miR-3473b Targeting NF-κB on Lung Cancer Cell Lung Metastasis and B Cell Number

作　　者：李冯洋 LI Fengyang(Zhumadian Central Hospital,Zhumadian,463000)

机构地区：[1]河南省驻马店市中心医院,463000

出　　处：《实用癌症杂志》2023年第6期871-875,共5页The Practical Journal of Cancer

摘　　要：目的探讨外泌体介导miR-3473b靶向NF-κB对肺癌细胞肺内转移及B细胞数量的影响。方法提取小鼠肺癌细胞(LLC)外泌体,鉴定外泌体形态及标志物表达。将Cy5.5标记的LLC来源外泌体与成纤维细胞共培养,共聚焦显微镜观察外泌体行踪。RT-PCR检测细胞内及外泌体内miR-3473b表达。分析LLC来源外泌体对小鼠肺内转移及B细胞数量的影响。蛋白免疫印迹法LLC来源外泌体miR-3473b对肺成纤维细胞NF-κB通路的影响。结果LLC来源外泌体微囊泡直径为30~100 nm,外泌体标记蛋白Hsp90、CD63呈阳性表达;LLC来源外泌体内miR-3473b表达水平显著高于细胞内(P<0.01);LLC来源外泌体组小鼠肺转移数目及面积显著多于对照组(P<0.05);LLC来源外泌体+miR-3473b抑制剂组小鼠肺转移数目及面积显著低于LLC来源外泌体组(P<0.05);LLC来源外泌体组小鼠肺内CD45+标记免疫细胞中B细胞百分比显著高于对照组(P<0.05);LLC来源外泌体+miR-3473b抑制剂组小鼠肺内CD45+标记免疫细胞中B细胞百分比显著低于LLC来源外泌体组(P<0.05);LLC来源外泌体组小鼠成纤维细胞p65和p-p65水平显著高于对照组,Nfkbid水平显著低于对照组(P<0.05);LLC来源外泌体+miR-3473b抑制剂组小鼠成纤维细胞p65和p-p65水平显著低于LLC来源外泌体组,Nfkbid水平显著高于对照组(P<0.05)。结论外泌体介导miR-3473b可能通过抑制肺成纤维细胞Nfkbid表达,靶向激活NF-κB信号通路,促进肺癌细胞肺内转移以及诱导肺内B细胞聚集。Objective To investigate the effect of exosomes-mediated miR-3473b targeting NF-κB on lung cancer cell lung metastasis and B cell number.Methods The exosomes of Lewis lung carcinoma(LLC)cells were extracted,and the morphology and marker expression of the exosomes were identified.Cy5.5 labeled LLC derived exosomes were co cultured with fibroblasts,and the exosomes were observed by confocal microscopy.The expression of miR-3473b was detected by RT-PCR.Effect of LLC derived exosomes on lung metastasis and the number of B cells in mice was analyzed.The effect of miR-3473b on NF-κB pathway in lung fibroblasts was detected by Western blot.Results The diameter of LLC derived exosomes was 30~100nm,with positive expression of HSP90 and CD63.The expression level of miR-3473b in LLC derived exocrine was significantly higher than that in cells(P<0.05).The number and area of lung metastasis in LLC derived exosomes group were significantly higher than those in the control group(P<0.05).The number and area of lung metastases in LLC derived exosomes+miR-3473b inhibitor group were significantly lower than those in LLC derived exosomes group(P<0.05).The percentage of B cells in lung CD45+labeled immune cells in LLC derived exosomes group was significantly higher than that in the control group(P<0.05).The percentage of B cells in lung CD45+labeled immune cells in LLC+miR-3473b inhibitor group was significantly lower than that in LLC derived exosomes group(P<0.05).The levels of p65 and p-p65 in LLC derived exosomes group were significantly higher than those in the control group,and the Nfkbid level was significantly lower than that in the control group(P<0.05);The levels of p65 and p-p65 in LLC derived exosomes+miR-3473b inhibitor group were significantly lower than those in LLC derived exosomes group,and Nfkbid level was significantly higher than that in the control group(P<0.05).Conclusion Exosomes-media-ted miR-3473b may inhibit lung fibroblast Nfkbid expression and target activation of NF-κB signaling pathway to promote lung

关 键 词：肺癌 肿瘤细胞源性外泌体 miR-3473b NF-κB信号通路 成纤维细胞 肺内转移 B细胞 

分 类 号：R734.2[医药卫生—肿瘤]