肺癌细胞外泌体miR-24-3p调控CD8^(+)T细胞抗肿瘤的功能及机制研究  

Study on the function and mechanism of lung cancer cells exosomal miR-24-3p regulating the anti-tumor of CD8^(+) T cell

作　　者：宋明磊 曹富民[1] 邢晓英 高立平[1] 井洪家 SONG Minglei;CAO Fumin;XING Xiaoying;GAO Liping;JING Hongjia(Department of Thoracic Surgery,the Fourth Hospital of Hebei Medical University,Hebei Shijiazhuang 050011,China;General Medical Department,the Second Hospital of Hebei Medical University,Hebei Shijiazhuang 050011,China;Department of Cardiothoracic Surgery,Chengde Central Hospital,Hebei Chengde 067000,China)

机构地区：[1]河北医科大学第四医院胸外科,河北石家庄050011 [2]河北医科大学第二医院全科医疗科,河北石家庄050011 [3]承德市中心医院心胸外科,河北承德067000

出　　处：《现代肿瘤医学》2023年第9期1601-1607,共7页Journal of Modern Oncology

基　　金：河北省重点科技研究计划(编号:20180554)。

摘　　要：目的:探究肺癌细胞外泌体miR-24-3p调控CD8^(+)T细胞抗肿瘤的功能及机制。方法:将16HBE exo、A549 exo、H522 exo、H460 exo、miR-NC exo、miR-24-3p exo与CD8^(+)T细胞共孵育(16HBE exo组、A549 exo组、H522 exo组、H460 exo组、miR-NC exo组、miR-24-3p exo组),与PBS共孵育组(PBS组)作为对照,CD8^(+)T与miR-24-3p exo共孵育后转染FGF11质粒(miR-24-3p exo+FGF11组)。CCK8法检测CD8^(+)T细胞的增殖能力。双荧光素酶报告基因实验验证miR-24-3p和FGF11的靶向关系。将各组CD8^(+)T细胞与A549细胞以20∶1的比例共孵育4 h(16HBE exo/CD8^(+)T组、A549 exo/CD8^(+)T组、H522 exo/CD8^(+)T组、H460 exo/CD8^(+)T组、miR-NC exo/CD8^(+)T组、miR-24-3p exo/CD8^(+)T组、PBS/CD8^(+)T组、miR-24-3p exo+FGF11/CD8^(+)T组),Elisa检测共孵育后细胞上清中INF-γ和IL-2的浓度,LDH法检测CD8^(+)T细胞对A549细胞的杀伤能力。结果:相比于PBS组,A549 exo组、H522 exo组、H460 exo组CD8^(+)T细胞增殖能力显著降低(P<0.05)。A549 exo/CD8^(+)T组、H522 exo/CD8^(+)T组、H460 exo/CD8^(+)T组细胞上清中INF-γ和IL-2的浓度显著下降(P<0.001),CD8^(+)T细胞杀伤能力亦显著下降(P<0.001)。双荧光素报告基因检测结果显示FGF11为miR-24-3p的靶基因。miR-24-3p exo组CD8^(+)T细胞增殖能力显著低于miR-NC exo组,miR-24-3p exo+FGF11组CD8^(+)T细胞增殖能力显著高于miR-24-3p exo组(P<0.05)。miR-24-3p exo/CD8^(+)T组细胞上清中INF-γ和IL-2浓度及CD8^(+)T细胞杀伤能力均显著低于miR-NC exo/CD8^(+)T组(P<0.001), miR-24-3p exo+FGF11/CD8^(+)T组细胞上清中INF-γ和IL-2浓度及CD8^(+)T细胞杀伤能力显著高于miR-24-3p exo/CD8^(+)T组(P<0.001)。结论:肺癌细胞外泌体miR-24-3p可通过靶向抑制FGF11而抑制CD8^(+)T细胞的抗肿瘤功能。Objective:To explore the function and mechanism of lung cancer cells exosomal miR-24-3p on the anti-tumor function of CD8^(+)T cells.Methods:16HBE exo,A549 exo,H522 exo,H460 exo,miR-NC exo,miR-24-3p exo and PBS were co-incubated with CD8^(+)T cells(16HBE exo group,A549 exo group,H522 exo group,H460 exo group,miR-NC exo group,miR-24-3p exo group,PBS group),CD8^(+)T was co-incubated with miR-24-3p exo and transfected into FGF11 plasmid(miR-24-3p exo+FGF11 group).The proliferation ability of CD8^(+)T cells was detected by CCK8 method.The dual-luciferase reporter gene experiment verified the targeting relationship between miR-24-3p and FGF11.The above groups of CD8^(+)T cells and A549 cells were co-incubated for 4 h at a ratio of 20∶1(16HBE exo/CD8^(+)T group,A549 exo/CD8^(+)T group,H522 exo/CD8^(+)T group,H460 exo/CD8^(+)T group,miR-NC exo/CD8^(+)T group,miR-24-3p exo/CD8^(+)T group,PBS/CD8^(+)T group,miR-24-3p exo+FGF11/CD8^(+)T group).Elisa was used to detect the concentrations of INF-γand IL-2 in the supernatant,and the killing ability of CD8^(+)T cells in each group was detected by LDH method.Results:Compared with the PBS group,the proliferation ability of CD8^(+)T cells in the A549 exo group,H522 exo group and H460 exo group was significantly decreased(P<0.05).The concentrations of INF-γand IL-2 in the supernatant of A549 exo/CD8^(+)T group,H522 exo/CD8^(+)T group and H460 exo/CD8^(+)T group were significantly decreased(P<0.001),and the killing ability of CD8^(+)T cell was also significantly decreased(P<0.001).The results of dual fluorescein reporter gene assay showed that FGF11 was the target gene of miR-24-3p.The proliferation ability of CD8^(+)T cells in miR-24-3p exo group was significantly lower than that in miR-NC exo group,and the proliferation ability of CD8^(+)T cells in miR-24-3p exo+FGF11 group was significantly higher than that in miR-24-3p exo group(P<0.05).The concentrations of INF-γand IL-2 in the supernatant and CD8^(+)T cell killing ability of the miR-24-3p exo/CD8^(+)T group were signific

关 键 词：肺癌 外泌体 miR-24-3p CD8^(+)T细胞 

分 类 号：R734.2[医药卫生—肿瘤]