摘要
目的:探讨IL-18在口腔鳞癌(OSCC)患者肿瘤组织中的表达水平及肿瘤相关巨噬细胞(TAMs)对OSCC肺转移病灶形成的影响。方法:本研究共纳入2015年1月至2017年12月我院收治的38例OSCC患者,另选取47例同期在我院体检中心进行体检的健康志愿者作为对照。ELISA法检测各组研究对象血清IL-18水平;免疫组化和Western blot分析OSCC患者肿瘤组织和癌旁组织中IL-18表达。建立小鼠肺转移模型,采用慢病毒转染法构建OSCC Tca8113 IL-18高表达细胞系(Tca8113-IL-18),Western blot检测IL-18、AKT、P-AKT、mTOR和P-mTOR表达。裸鼠尾静脉注射Tca8113与Tca8113-IL-18细胞,45 d后统计小鼠肺组织转移病灶数,CD11b和F4/80标记后流式细胞术分选巨噬细胞;ELISA检测小鼠血清中CXCL9、CXCL10、CSF-1和CCL-2表达;qRT-PCR检测小鼠肿瘤组织CXCL9、CXCL10、CSF-1和CCL-2表达;Western blot检测CXCL9、CXCL10、AKT、P-AKT、mTOR和P-mTOR表达。结果:OSCC患者血清中IL-18表达显著高于健康志愿者,肿瘤组织中IL-18表达显著高于癌旁组织(P<0.05)。IL-18在Tca8113-IL-18高表达,成功构建高表达细胞系,P-AKT和P-mTOR在Tca8113-IL-18细胞系中表达高于Tca8113细胞。IL-18可显著增加Tca8113在裸鼠肺部形成转移病灶数(P<0.05),Tca8113-IL-18组裸鼠肺部转移瘤数明显高于Tca8113组。流式细胞术分选结果显示,Tca8113-IL-18组裸鼠肿瘤组织中TAMs数明显增多。ELISA结果显示Tca8113-IL-18裸鼠血清中CXCL9和CXCL10表达明显低于Tca8113组,而CSF-1和CCL-2表达明显升高。qRT-PCR和Western blot显示Tca8113-IL-18肿瘤组织中CXCL9和CXCL10表达明显低于Tca8113组,而CSF-1和CCL-2表达明显增加,Tca8113-IL-18组P-AKT和P-mTOR表达高于Tca8113组。结论:OSCC患者血清和肿瘤组织中IL-18表达显著升高,IL-18可能通过激活PI3K/AKT/mTOR信号通路下调CXCL9、CXCL10表达,从而募集TAMs促进OSCC肺部转移病灶形成。
Objective:To investigate expression of IL-18 in tumor tissues of patients with oral squamous cell carcinoma(OSCC)and effect of tumor-associated macrophages(TAMs)on formation of lung metastases in OSCC.Methods:38 patients with OSCC admitted to our hospital from January 2015 to December 2017 were included,another 47 healthy people who underwent physical examination at physical examination center of our hospital were selected as controls. Serum IL-18 level was measured by ELISA of each group. IL-18 expression in tumor tissues and adjacent tissues of patients with OSCC was detected by immunohistochemistry and Western blot. A mouse lung metastasis model was established. IL-18 high-expression cell line of oral OSCC Tca8113 was constructed by lentiviral transfection(Tca8113-IL-18). Expressions of IL-18,AKT,P-AKT,mTOR and P-mTOR were detected by Western blot. Nude mice were injected Tca8113 and Tca8113-IL-18 cells into tail vein,number of metastatic lesions in lung tissue was analyzed at 45 d.Macrophages were sorted by flow cytometry marked with CD11 b and F4/80. Expressions of CXCL9,CXCL10,CSF-1 and CCL-2 in serum were detected by ELISA. Expressions of CXCL9,CXCL10,CSF-1 and CCL-2 in tumor tissue were detected by qRT-PCR. CXCL9,CXCL10,AKT,P-AKT,mTOR and P-mTOR expressions were detected by Western blot.Results:Expression of IL-18 in serum of patients with OSCC was significantly higher than that in healthy volunteers,and expressions of IL-18,CXCL9 and CXCL10 in tumor tissues were significantly higher than those in adjacent tissues(P<0.05). IL-18 was highly expressed in Tca8113-IL-18 cell line,high expression of cell line was successfully constructed,P-AKT and P-mTOR expression were higher in Tca8113-IL-18 cell line than Tca8113 cells. IL-18 significantly increased number of Tca8113 metastatic lesions in lungs of mice(P<0.05),number of lung metastases in Tca8113-IL-18 group was significantly higher than that in Tca8113 group. Flow cytometry showed that number of TAMs in tumor tissues of Tca8113-IL-18 mice increased significantly. ELISA showed that expressions of CXCL9 and CXCL10 in Tca8113-IL-18 mice was significantly lower than that in Tca8113 group,while expressions of CSF-1 and CCL-2 were significantly increased. qRT-PCR and Western blot showed that expressions of CXCL9 and CXCL10 in Tca8113-IL-18 tumor tissues were significantly lower than that in Tca8113 group,while expressions of CSF-1 and CCL-2 were significantly increased. Expression of P-AKT and P-mTOR in Tca8113-IL-18 was higher than that in Tca8113 group.Conclusion:IL-18 levels in serum and tumor tissues of patients with OSCC are significantly increased. IL-18 may down-regulate expressions of CXCL9 and CXCL10 by activating PI3 K/AKT/mTOR signaling pathway to recruit TAMs to promote formation of OSCC lung metastatic lesions.
作者
周卫
曾华兴
肖才文
ZHOU Wei;ZENG Hua-Xing;XIAO Cai-Wen(Department of Otolaryngology Head and Neck Surgery,Affiliated Hospital of Jianghan University,the Sixth Hospital of Wuhan,Wuhan 430056,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2021年第10期1221-1225,共5页
Chinese Journal of Immunology
