微小RNA-148a-3p对肝癌树突状细胞活化的影响及其机制  

Effect and potential mechanism of microRNA-148a-3p on the activation of dendritic cells in hepato cellularcarcinoma

作　　者：宋晓静[1] 丁方回 骆伟 倪海旭 李汛[1] Song Xiaojing;Ding Fanghui;Luo Wei;Ni Haixu;Li Xun(Department of General Surgery,the First Hospital of Lanzhou University,Lanzhou 730000,China)

机构地区：[1]兰州大学第一医院普外科,兰州730000

出　　处：《中华实验外科杂志》2023年第5期855-858,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金地区项目(82060119);兰州市科技局科技发展指导计划项目(2019-ZD-35);兰州大学第一医院院内基金项目(ldyy2018-58)。

摘　　要：目的探讨微小RNA(miR)-148a-3p对肝癌树突状细胞活化的影响及其机制。方法分离肝癌患者外周血树突状细胞,通过双荧光素酶报告验证miR-148a-3p和细胞因子信号转导抑制因子1(SOCS1)的靶向关系。树突状细胞分为4组:对照组、miR-148a-3p组、SOCS1组和miR-148a-3p+SOCS1组。通过转染miR-148a-3p类似物和/或SOCS1质粒来实现过表达。检测各组细胞中miR-148a-3p和SOCS1蛋白、抗原呈递蛋白(CD11c和CD86)、白细胞介素-6(IL-6)、干扰素-α(IFN-α)的表达水平和树突细胞迁移能力。多组间的差异采用单因素方差分析(ANOVA),组间两两比较采用SNK-q检验。结果miR-148a-3p组的miR-148a-3p水平高于对照组(4.21±0.38比1.00±0.07,t=66.461,P<0.05),SOCS1组的SOCS1蛋白表达水平显著高于对照组(2.91±0.25比1.00±0.08,t=58.212,P<0.05)。miR-148a-3p+SOCS1组的SOCS1蛋白水平显著高于miR-148a-3p组(1.06±0.10比0.24±0.03,t=62.883,P<0.05);且显著低于SOCS1组(1.06±0.10比2.91±0.25,t=54.966,P<0.05)。miR-148a-3p组的CD11(3.24±0.27比1.00±0.08)、CD86(2.96±0.25比1.00±0.07)、IL-6[(517.34±48.63)pg/ml比(365.72±31.25)pg/ml]、IFN-α[(72.91±7.09)pg/ml比(48.34±4.24)pg/ml]水平显著高于对照组(t=63.636、60.397、20.984、23.793,P<0.05),SOCS1组的CD11c、CD86、IL-6和IFN-α蛋白水平显著低于对照组(P<0.05)。miR-148a-3p+SOCS1组的CD11(1.35±0.11比3.24±0.27、0.21±0.02)、CD86(0.98±0.09比2.96±0.25、0.23±0.03)、IL-6[(371.06±35.98)pg/ml比(517.34±48.63)、(278.85±26.44)pg/ml]、IFN-α[(49.28±5.06)pg/ml比(72.91±7.09)、(30.56±3.21)pg/ml]水平显著低于miR-148a-3p组且显著高于SOCS1组(t=51.861、81.572、59.615、78.651、19.345、16.347、21.703、24.992,P<0.05)。miR-148a-3p组迁移能力[(42.96±6.01)%比(28.33±4.25)%]显著高于对照组(t=15.900,P<0.05);SOCS1组的迁移能力[(16.35±2.73)%比(28.33±4.25)%]显著低于对照组(t=18.973,P<0.05);miR-148a-3p+SOCS1组的迁移能力[(30.95±4.51)%比(42.96±6.01)%、(16.35±2.73)%]显著�Objective To explore the effect and potential mechanism of microRNA(miR)-148a-3p on the activation of dendritic cells in hepatocellular carcinoma(HCC).Methods Dendritic cells were iso-lated from peripheral blood of patients with HCC.The targeting relationship between miR-148a-3p and suppressors of cytokine signaling 1(SOCSI)was verified by dual luciferase reporter gene assay.The cultured dendritic cells were divided into control group,miR-148a-3p group,SOCS1 group and miR-148a-3p+SOCS1 group.Cells were transfected with miR-148a-3p mimics and/or SOCS1 plasmid to establish overex-pression models.The expression levels of miR-148a-3p and SOCS1 proteins,membrane proteins provided by antigen-presenting cells(CD11c and CD86),interleukin-6(IL-6),interferon-α(IFN-α)and dendritic cell migration ability were measured.Results Expression level of miR-148a-3p was(4.21±0.38)in miR-148a-3p group,which was higher than(1.00±0.07)in control group(t=66.461,P<0.05),and the protein expression level of SOCS1 was(2.91±0.25)in SOCS1 group,significantly higher than(1.00±0.08)in control group(t=58.212,P<0.05).The protein expression level of SOCS1 was(1.06±0.10)in miR-148a-3p+SOCS1 group,significantly higher than(0.24±0.03)in miR-148a-3p group(t=62.883,P<0.05),and significantly lower than(2.91±0.25)in SOCS1 group(t=54.966,P<0.05).Levels of CD11,CD86,IL-6 and IFN-αwere(3.24±0.27),(2.96±0.25),(517.34±48.63)and(72.91±7.09)pg/ml respectively in miR-148a-3p group,which were higher than(1.00±0.08),(1.00±0.07),(365.72±31.25)and(48.34±4.24)pg/ml in control group(t=63.636,60.397,20.984,23.793,P<0.05).Levels of CD11,CD86,IL-6 and IFN-αin SOCS1 group were significantly higher than those in control group(P<0.05).Levels of CD11,CD86,IL-6 and IFN-αwere(1.35±0.11),(0.98±0.09),(371.06±35.98)and(49.28±5.06)pg/ml in miR-148a-3p+S0CS1 group,which were lower than(3.24±0.27),(2.96±0.25),(517.34±48.63),and(72.91±7.09)pg/ml in miR-148a-3p group,and higher than(0.21±0.02),(2.96±0.25),(278.85±26.44)and(30.56±3.21)pg/ml in S0CS1 group

关 键 词：肝癌 树突状细胞 细胞因子信号转导抑制因子1 

分 类 号：R735.7[医药卫生—肿瘤]