【摘要】 目的:探讨肿瘤相关巨噬细胞（TAMs）来源的白细胞介素6 （IL-6）对口腔鳞状细胞癌（OSCC） Cal27细胞中白血病抑制因子（LIF）表达及侵袭和迁移的影响,并阐明其可能的作用机制。方法:体外培养人OSCC Cal27细胞和小鼠腹腔巨噬细胞（Raw264.7细胞）。Raw264.7细胞分为对照组和实验组,对照组细胞用普通培养基孵育,实验组细胞用Cal27细胞上清液孵育得到TAMs,实时荧光定量PCR （RT-qPCR）法和酶联免疫吸附测定（ELISA）法检测2组细胞中IL-6 mRNA表达水平及IL-6水平。Cal27细胞分为对照组和实验组(5μg·L^-1Tocilizumab),对照组直接加入TAMs上清液,实验组提前用IL-6受体（IL-6R）抑制剂Tocilizumab干预后加入TAMs上清液,Transwell小室实验检测2组Cal27细胞迁移和侵袭能力,细胞划痕实验检测2组Cal27细胞划痕愈合率,RT-qPCR法和Western blotting法检测2组Cal27细胞中LIF mRNA及蛋白表达水平。结果:RT-qPCR法和ELISA法检测,与对照组Raw264.7细胞比较,实验组TAMs中IL-6 mRNA表达水平和IL-6水平明显升高（P<0.01）。Transwell小室实验,与对照组比较,实验组Cal27细胞迁移和侵袭细胞数明显减少（P<0.01）。细胞划痕实验,与对照组比较,实验组Cal27细胞划痕愈合率明显降低（P<0.01）。RT-qPCR法和Western blotting法检测,与对照组比较,实验组Cal27细胞中LIF mRNA和蛋白表达水平明显降低（P<0.01）。结论:TAMs来源的IL-6通过上调肿瘤细胞中LIF表达进而促进OSCC Cal27细胞的侵袭和迁移。更多还原

【Abstract】 Background:To investigate the effects of tumor associated macrophages（TAMs）-derived interleukin-6（IL-6）on the leukemia inhibitory factor（LIF）expression in oral squamous cell carcinoma（OSCC） Cal27 cells and the invasion and migration of OSCC Cal27 cells,and to clarify their possible mechanisms. Methods:The human OSCC Cal27 cells and mouse peritoneal macrophages（Raw264. 7 cells）were cultured in υitro. The Raw264. 7 cells were divided into control and experimental groups. The cells in control group were incubated with DMEM,and the cells in experimental group were incubated with Cal27 conditioned medium（Cal27-CM）to obtain the TAMs. The expression levels of IL-6 mRNA and IL-6 levels in the cells in two groups were detected by real-time fluorescence quantitative PCR（RT-qPCR）and enzyme-linked immunosorbent assay（ELISA） methods. The Cal27 cells were divided into control group and experimental group(5 μg·L^-1Tocilizumab). The Cal27 cells in control group were directly treated with TAMs conditioned medium,and the Cal27 cells in experimental group were treated with interleukin 6 receptor（IL-6R）inhibitor Tocilizumab in advance and then treated with TAMs conditioned medium. Transwell chamber experiment was used to detect the migration and invasion abilities of Cal27 cells in two groups. Cell scratch assay was used to detect the scratch healing rates of Cal27 cells in two groups. RT-qPCR and Western blotting methods were used to detect the expression of levels LIF mRNA and protein in Cal27 cells in two groups. Results:The results of RT-qPCR and ELISA showed that the mRNA expression level of IL-6 and the IL-6 level in the TAMs in experimental group were significantly higher than those in the RAW264. 7 cells in control group（P<0. 01）. The results of Transwell chamber experiment showed that compared with control group,the number of migration and invasion cells in experimental group was significantly reduced（P<0. 01）. The results of cell scratch experiment showed that compared with control group,the scratch healing rate of the Cal27 cells in experimental group was significantly decreased（P<0. 01）. The RT-qPCR and Western blotting results showed that compared with control group,the expression levels of LIF mRNA and protein in the Cal27 cells in experimental group were decreased down-regulated（P<0. 01）. Conclusion:TAMs-derived IL-6 promotes the invasion and migration of OSCC Cal27 cells by up-regulating the expression of LIF in tumor cells.更多还原