HMGB1藉由NF-κB通路诱导肺癌细胞增殖并抑制免疫细胞功能  

HMGB1 induces proliferation of lung cancer cells and suppresses immune cell function via NF-κB pathway

作　　者：王耀辉[1] 孟宇[2] 钱一可 陈文利 厉荣玉[1] 董博翰[1] WANG Yaohui;MENG Yu;QIAN Yike;CHEN Wenli;LI Rongyu;DONG Bohan(Department of Microbiology and Immunology,Wannan Medical College,Wuhu 241000,China;Department of Biochemistry,Wannan Medical College,Wuhu 241000,China;School of Pharmacy,Wannan Medical College,Wuhu 241000,China)

机构地区：[1]皖南医学院微生物学与免疫学教研室,芜湖241000 [2]皖南医学院生物化学教研室,芜湖241000 [3]皖南医学院药学院,芜湖241000

出　　处：《中国免疫学杂志》2025年第3期628-633,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金青年基金项目(81402351);安徽省教育厅自然科学研究项目(KJ2019A0415);安徽省高校优秀青年人才支持计划(重点)(gxyqZD2019040);皖南医学院学术和技术带头人后备人选;皖南医学院2020年青年卓越人才培养项目(wyqnyx202001)。

摘　　要：目的:探究肺癌细胞HMGB1通过NF-κB通路影响肺癌细胞本身和免疫细胞功能的分子机制。方法:Western blot检测LLC细胞、Raw264.7细胞、小鼠脾细胞中的HMGB1表达,同时用甘草酸(GA)抑制肺癌细胞中HMGB1表达,制备低HMGB1的肿瘤细胞裂解物(TCL);流式细胞术和CCK-8检测内源性HMGB1抑制或低HMGB1的TCL对肺癌细胞凋亡和增殖的影响;通过反复冻融的方法制备正常HMGB1的TCL或低HMGB1的TCL,然后用其干预Raw264.7细胞和小鼠脾细胞48 h,采用流式细胞术检测细胞凋亡和CD69表达,并采用ELISA检测细胞因子IL-2、IL-4、IL-6、TNF-α和TNF-β的分泌;Western blot检测肺癌细胞或免疫细胞中NF-κB信号通路关键信号分子的蛋白表达。结果:HMGB1在LLC细胞、Raw264.7细胞、小鼠脾细胞中都有表达,其中,LLC细胞HMGB1的表达最高,并且30μg/ml的GA对LLC细胞中HMGB1表达的抑制效果最佳。LLC细胞内源性HMGB1可以促进细胞增殖。TCL中外源的HMGB1则诱导肺癌细胞凋亡,抑制免疫细胞活化增殖。肺癌细胞中内源性HMGB1的抑制会导致NF-κB信号通路中凋亡诱导因子CASP9的激活,而在低HMGB1的TCL作用后的肺癌细胞或免疫细胞中,CASP9会被抑制。结论:肿瘤细胞HMGB1对肺癌的发生具有双重作用,促进肺癌细胞增殖的同时抑制免疫细胞的功能,进而引起肺癌的发生,该过程与不同细胞中NF-κB信号通路的激活有关。Objective:To investigate the molecular mechanisms by which HMGB1 in lung cancer cells affects the function of lung cancer cells themselves and immune cells through the NF-κB pathway.Methods:Western blot detected HMGB1 expressions in Lewis lung cancer(LLC)cells,Raw264.7 cells,and mouse spleen cells,while tumor cell lysates(TCL)with low HMGB1 was prepared by inhibiting HMGB1 expression in lung cancer cells with glycyrrhetinic acid(GA);the effects of endogenous HMGB1 inhibition or TCL with low HMGB1 on apoptosis and proliferation of lung cancer cells were detected by flow cytometry and CCK-8;TCL with normal HMGB1 or TCL with low HMGB1 was prepared by freeze-thawing;Raw264.7 cells and mouse splenocytes were treated with them for 48 h.Apoptosis and CD69 expression were detected by flow cytometry,and secretion of cytokines IL-2,IL-4,IL-6,TNF-α and TNF-β were detected by ELISA;Western blot detected lung cancer cells or immune cells.Western blot was performed to detect the protein expression of key signaling molecules of the NF-κB signaling pathway in lung cancer cells or immune cells.Results:HMGB1 was expressed in LLC cells,Raw264.7 cells,and mouse spleen cells,among which LLC cells had the highest expression of HMGB1,and 30μg/ml GA had the best inhibitory effect on HMGB1 expression in LLC cells.Endogenous HMGB1 in LLC cells could promote cell proliferation.Exogenous HMGB1 in TCL induced apoptosis in lung cancer cells and inhibited immune cell activation and proliferation.Inhibition of endogenous HMGB1 in lung cancer cells leaded to activation of the apoptosis-inducing factor CASP9 in the NF-κB signaling pathway,which was inhibited in lung cancer cells or immune cells after the action of TCL with low HMGB1.Conclusion:Tumor cell HMGB1 has a dual role in lung carcinogenesis,promoting the proliferation of lung cancer cells while suppressing the function of immune cells,which in turn causes lung carcinogenesis,a process associated with the activation of the NF-κB signaling pathway in different cells.

关 键 词：抗肿瘤免疫 细胞凋亡 NF-κB信号通路 巨噬细胞 小鼠脾细胞 肺癌 

分 类 号：R734.2[医药卫生—肿瘤]