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目的观察IL-6诱导小鼠髓源性DCs分化成熟障碍过程中SOCS3的表达变化并探讨其分子机制。方法分离小鼠骨髓细胞,采用黏附法结合GM-CSF和IL-4刺激法在体外培养髓源性DCs(BMDCs)。用LPS诱导iDCs成熟,采用Real-Time RT-PCR和Western blot方法,观察IL-6处理前后LPS诱导的BMDCs中SOCS3表达的变化,并用甲基化特异性PCR观察SOCS3启动子CpG岛的甲基化状态。结果 Real-Time RT-PCR和Western blot结果显示,LPS能显著诱导BMDCs SOCS3 mRNA和蛋白的表达上调;而经过IL-6处理后,LPS诱导BMDCs SOCS3 mRNA和蛋白表达上调的能力被显著抑制(P<0.05)。甲基化特异性PCR与单独用LPS处理的BMDCs相比,经IL-6处理后的BMDCs SOCS3启动子区CpG岛被甲基化。结论 IL-6通过诱导SOCS3基因启动子区CpG岛的甲基化来抑制SOCS3表达,进而阻遏iDCs分化成熟,促进胃癌等肿瘤细胞侵袭转移。
Objective To investigate the changes of SOCS3 expression and the molecular mechanism of IL-6-induced maturation of myeloid DCs in mice. Methods Mouse bone marrow cells were isolated and myeloid DCs (BMDCs) were cultured in vitro by adhesion method combined with GM-CSF and IL-4 stimulation. Real-time RT-PCR and Western blot were used to observe the changes of SOCS3 expression in LPS-induced BMDCs before and after IL-6 treatment. The methylation-specific PCR was used to observe the methylation of SOCS3 promoter in CpG island State of affairs Results The results of Real-Time RT-PCR and Western blot showed that LPS significantly up-regulated the expression of SOCS3 mRNA and protein in BMDCs. The ability of LPS-induced SOCS3 mRNA and protein expression in BMDCs was significantly inhibited by IL-6 treatment (P <0.05). Methylation-Specific PCR CpG islands in the SOCS3 promoter region of BMDCs treated with IL-6 were methylated compared to BMDCs treated with LPS alone. Conclusion IL-6 inhibits the expression of SOCS3 by inducing methylation of CpG island in SOCS3 promoter region, thereby inhibiting the differentiation and maturation of iDCs and promoting the invasion and metastasis of gastric cancer cells.