TNF-α通过NF-κB增强Th9细胞分化抑制肺癌细胞生长  

TNF-αenhances Th9 cell differentiation and inhibits lung cancer growth through NF-κB

作　　者：李彦 顾泽鑫[2] 孟祥茹 兰冰 李静龙 李翠翠 LI Yan;GU Zexin;MENG Xiangru;LAN Bing;LI Jinglong;LI Cuicui(Department of Respiratory Medicine,Second Affiliated Hospital of Qiqihar Medical College,Qiqihar 161000,China;Section 2 of Respiratory Department,Second Affiliated Hospital of Qiqihar Medical College,Qiqihar 161000,China;Department of Thoracic Surgery,Second Affiliated Hospital of Qiqihar Medical College,Qiqihar 161000,China)

机构地区：[1]齐齐哈尔医学院附属第二医院呼吸内一科,161000 [2]齐齐哈尔医学院附属第二医院呼吸二科,161000 [3]齐齐哈尔医学院附属第二医院胸外科,161000

出　　处：《免疫学杂志》2022年第3期228-233,共6页Immunological Journal

基　　金：2019年度黑龙江省省属高等学校基本科研业务费科研项目(2019-KYYWF-1211)。

摘　　要：目的研究TNF-α通过NF-κB增强Th9细胞分化抑制肺癌细胞生长的功能并对其相关机制进行初步探讨。方法分离野生型小鼠脾脏CD4^(+)T细胞,体外诱导Th9细胞分化。荧光定量PCR检测IL-9、IL-5和IL-13表达,ELISA检测Th细胞分泌IL-9变化。荧光定量PCR检测TNF-α在不同时间点对Th9细胞中IL-9表达的影响;TNF-α刺激分化的Th9细胞,免疫印迹检测NF-κB通路相关蛋白p-IKKα/β和IkB-α表达。使用NF-κB通路阻断剂,荧光定量PCR检测IL-9、IL-5和IL-13的表达。将体外TNF-α刺激诱导分化的Th9淋巴细胞通过尾静脉注射入肺癌小鼠体内,荧光定量PCR检测小鼠脾脏中IL-9、IL-5和IL-13的表达,免疫印迹检测小鼠肺癌组织中凋亡蛋白Bax/Bcl2表达,免疫荧光染色检测小鼠肺癌组织细胞增殖情况。结果荧光定量PCR显示,TNF-α联合Th9极化细胞因子TGF-β和IL-4增加Th细胞中IL-9、IL-5和IL-13的表达;ELISA实验结果显示,Th细胞分泌IL-9明显增多;荧光定量PCR显示,TNF-α处理后的Th9细胞中IL-9的表达在第2天或第3天达到最高水平;免疫印迹实验显示,TNF-α刺激NF-κB通路相关蛋白p-IKKα/β和IkB-α表达;荧光定量PCR显示,NF-κB通路阻断剂可消除TNF-α诱导的IL-9、IL-5和IL-13的表达。最后,荧光定量PCR显示,Th9淋巴细胞通过尾静脉注射入肺癌小鼠体内后,小鼠脾脏中IL-9、IL-5和IL-13的表达明显升高,免疫印迹显示小鼠肺癌组织中凋亡蛋白Bax/Bcl2表达升高,免疫荧光染色发现细胞增殖减少。结论TNF-α可以通过NF-κB信号通路增强Th9细胞分化增强肺癌细胞凋亡,抑制肺癌细胞增殖。This study was designed to investigate the inhibitory effect of TNF-α on the growth of lung cancer cells by enhancing the differentiation of Th9 cells through NF-κB and to explore the related mechanism.CD4^(+) T cells were isolated from spleen of wild-type mice and induced to differentiation in vitro.The effect of TNF-α on IL-9 expression of Th9 cells at different time points was detected.Th9 cells were stimulated with TNF-α to block NF-κB signaling pathway,and the expression of IL-9,IL-5 and IL-13 induced by TNF-α were detected by fluorescence quantitative PCR.TNF-α-stimulated Th9 lymphocytes were injected into the tail vein of lung cancer mice in vitro to detect the expression of IL-9,IL-5 and IL-13,as well as the expression of apoptosis and proliferation related proteins in vitro.Data showed thatTNF-αcombined with Th9 polarizing cytokines TGF-β and IL-4 increased the expression of IL-9,IL-5 and IL-13 in Th cells.The expression of IL-9 in Th9 cellsreached the highest level on day 2 or day 3 after TNF-α treatment;TNF-α stimulated the expression of NF-κB pathway related proteins p-IKKα/β and IkB-αin differentiated Th9 cells.NF-κB pathway blockers could eliminateTNF-α-induced upregulation of IL-9,IL-5,IL-13.After differentiated Th9 lymphocytes injection,the expressionof IL-9,IL-5 and IL-13 in spleen of mice were significantly increased,the expression of apoptotic protein Bax/Bcl2 was increased while the cell proliferation was decreased in mouse lung cancer tissues.Taken together,TNF-α canenhance the differentiation of Th9 cells through NF-κB signaling pathway,enhance the apoptosis of lung cancercells,and inhibit the proliferation of lung cancer cells.

关 键 词：TNF-Α NF-ΚB Th9细胞分化 肺癌 

分 类 号：R734.2[医药卫生—肿瘤]