IL⁃18诱导的单核细胞THP⁃1对肺癌细胞增殖、迁移和侵袭的影响
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山东省自然科学基金(ZR201807060425)


Effects of IL⁃18⁃induced mononuclear cell THP⁃1 on proliferation,migration and invasion of lung cancer cells
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    目的:探讨外源性白介素18(interleukin 18,IL-18)对单核细胞THP-1的活化作用以及IL-18激活诱导的THP-1对肺癌A549细胞增殖、迁移和侵袭的影响。方法:通过IL-18体外诱导THP-1模拟激活体内肿瘤微环境中的肿瘤相关巨噬细胞(tumor associated macrophage,TAM)的作用。Transwell法检测IL-18对单核细胞THP-1的趋化作用;外源性IL-18刺激THP-1细胞后流式细胞术检测M1、M2型TAM的比例;IL-18激活诱导的THP-1和肺癌细胞A549共培养,分别以克隆形成实验、细胞划痕实验、细胞侵袭实验检测共培养后A549细胞增殖、迁移和侵袭的变化;扫描电镜观察共培养后A549细胞形态的变化;qRT-PCR和Western blot检测共培养后A549细胞上皮间质转化(epithelial mesenchymal transition,EMT)标志蛋白E-cadherin和N-cadherin、Snail以及Slug的表达。结果:IL-18对THP-1细胞具有显著的趋化作用;外源性IL-18刺激THP-1细胞表型向M2 TAM方向分化;IL-18诱导THP-1促进了A549细胞增殖、迁移和侵袭能力;扫描电镜结果显示IL-18诱导THP-1促进A549细胞伪足生长;qRT-PCR和Western blot检测结果显示经IL-18诱导的THP-1抑制A549细胞E-cadherin的表达,促进N-cadherin、Snail以及Slug的表达,说明IL-18诱导THP-1激活A549细胞发生EMT。结论:IL-18可以诱导THP-1并促进其活化,活化后的THP-1通过激活EMT机制促进肺癌A549细胞的增殖、迁移和侵袭。

    Abstract:

    Objective:To investigate the activation of monocytes(THP-1)by exogenous interleukin 18(IL-18)and the proliferation,metastasis and invasion of lung cancer A549 cells by THP-1 induced by IL-18 activation influences. Methods:The role of tumor associated macrophages(TAMs)in tumor microenvironment in vivo was simulated by IL-18 induced THP-1 in vitro. The chemotactic effect of IL-18 on mononuclear cell THP-1 was detected by Transwell method. Exogenous IL-18 stimulated THP-1 cells and detected the proportion of M1 and M2 TAMs;IL-18-activated TAMs and lung cancer cells A549 were co-cultured. The changes of proliferation,migration and invasion of co-cultured A549 cells were detected by cloning experiment,cell scratch test and cell invasion assay. The morphology of A549 cells after co-culture was observed by scanning electron microscope. The expressions of epithelial mesenchymal transition(EMT)marker proteins E-cadherin,N-cadherin,Snail and Slug in A549 cells were detected by qRT-PCR and Western blot. Results:IL-18 had a significant chemotactic effect on THP-1 cells. Exogenous IL-18 stimulated the THP-1 cell phenotype from M2 to TAMs. IL-18 induced THP-1 promoted the proliferation,migration and invasion of A549. Scanning electron microscopy showed that THP-1 promoted the growth of pseudopod of A549 cells. At the same time,the expression of E-cadherin was inhibited,and the expression of N-cadherin,Snail and Slug was promoted,indicating that IL-18 induced THP-1 activated EMT in A549 cells. Conclusion:In lung cancer,IL-18 can activate THP-1 and promote their activation. Activated THP-1 promotes the proliferation,migration and invasion of lung cancer A549 cells by activating EMT.

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聂兆超,石 星,毛海婷. IL⁃18诱导的单核细胞THP⁃1对肺癌细胞增殖、迁移和侵袭的影响[J].南京医科大学学报(自然科学版),2019,(10):1430-1436

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  • 收稿日期:2018-10-18
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  • 在线发布日期: 2019-11-01
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