LPS通过宫颈癌细胞来源的IL-6调节巨噬细胞分泌和趋化活性  

IL-6 expression promoted by LPS in cervical cancer cells regulates cytokine expression and recruitment of macrophages

作　　者：张春焕 阚士锋[1] 魏守娟 张之杰 Zhang Chunhuan;Kan Shifeng;Wei Shoujuan(Department of Clinical Laboratory,Qilu Hospital of Shandong University,Jinan 250012)

机构地区：[1]山东大学齐鲁医院检验科,济南250012 [2]山东省妇幼保健院超声科,济南250014

出　　处：《现代妇产科进展》2021年第12期905-909,共5页Progress in Obstetrics and Gynecology

摘　　要：目的探讨LPS对子宫颈癌细胞-巨噬细胞相互作用的影响和作用机制。方法LPS作用于HeLa细胞系,ELISA检测LPS对HeLa细胞细胞因子分泌的影响。以PMA于体外诱导THP-1来源巨噬细胞,在诱导分化过程中加经LPS处理后的HeLa细胞条件培养基,检测含有LPS的HeLa细胞条件培养基对巨噬细胞分泌谱的作用;Transwell实验检测LPS处理后HeLa细胞条件培养基对巨噬细胞募集作用。利用siRNA转染敲除HeLa细胞中IL-6的表达,观察IL-6敲除的HeLa细胞经LPS刺激后对巨噬细胞分泌和募集的作用。Western blot法检测经LPS刺激后HeLa细胞中NF-κB通路活化状态,验证NF-κB通路在LPS诱导HeLa细胞分泌IL-6中的作用。结果经LPS处理后HeLa细胞IL-6分泌水平显著上调;经LPS诱导的HeLa细胞上清显著增强巨噬细胞IL-1β、IL-6的分泌,并抑制巨噬细胞IL-10和CCL22的分泌;经LPS诱导的HeLa细胞上清对巨噬细胞的募集作用显著增强;HeLa细胞敲除IL-6可逆转LPS对巨噬细胞分泌和募集的调节作用;LPS激活HeLa细胞NF-κB通路活性,NF-κB通路抑制剂PDTC显著逆转LPS对HeLa细胞IL-6分泌的促进作用。结论LPS通过NF-κB通路促进子宫颈癌细胞IL-6的分泌;子宫颈癌细胞通过IL-6调节巨噬细胞的分泌活性及向肿瘤细胞的募集作用,提示LPS可能通过间接调节肿瘤浸润巨噬细胞的数量和功能,诱导肿瘤促炎微环境的形成。Objective:To investigate the role and mechanism of LPS in the interaction between cervical cancer cells and macrophages.Methods:HeLa cells were treated with LPS and their cytokine secretion profiles were analyzed by ELISA.The conditioned medium of cervical cancer cells after LPS treatment was collected and added during the differentiation process of THP-1 cells to macrophages induced by PMA,and the cytokine transcription and secretion of macrophages were analyzed.The recruitment of macrophages by LPS-treated HeLa cells was also analyzed using a transwell experiment.IL-6 siRNA was transfected into HeLa cells to explore its role in LPS-regulated secretion and recruitment of THP-1-derived macrophages.Finally,Western blot was performed to explore the role of NF-κB in LPS-promoted IL-6 expression by cervical cancer cells.Results:IL-6 was significantly up-regulated in HeLa cells after LPS treatment.LPS-stimulated supernatant of HeLa cells promoted secretion of M1-type cytokine,including IL-1βand IL-6,in THP-1-derived macrophages.However,it inhibited the expression of M2-type cytokine,including IL-10 and CCL22.IL-6 inhibition in HeLa cells by siRNA transfection reversed the effects of LPS-treated supernatant.The recruitment of macrophages by LPS-stimulated Hela cells supernatant was also enhanced and was significantly inhibited by IL-6 siRNA transfection.Finally,phosphorylation of the NF-κB signaling pathway in HeLa cells occurred quickly after LPS treatment.The NF-κB signaling pathway inhibitor,PDTC,significantly inhibited LPS-induced IL-6 expression.Conclusion:LPS promotes IL-6 secretion in cervical cancer cells through activation of the NF-κB pathway.The elevated IL-6 expression was involved in regulating the secretion profile of THP-1-derived macrophages,and promoted their recruitment by cervical cancer cells.These results suggest that LPS might involve in the regulation of cervical cancer cells on tumor-infiltrated macrophages,and contribute to pro-inflammatory tumor microenvironment.

关 键 词：脂多糖 宫颈癌细胞 巨噬细胞 白细胞介素-6 信号通路 

分 类 号：R737.3[医药卫生—肿瘤]