目的探讨微环境因子在转化生长因子β（TGF-β）诱导胃癌上皮-间充质转化（EMT）中的作用。方法建立了TGF-β诱导胃癌细胞发生EMT模型，利用实时定量PCR、免疫荧光染色技术检测了EMT相关指标的表达情况，采用Transwell迁移实验、侵袭实验、划痕修复实验检测了胃癌细胞的转移能力；同时，利用实时定量PCR（RT-qPCR）和ELISA检测了TGF-β诱导刺激后胃癌微环境因子的改变，并通过免疫印迹检测了其下游信号分子的活性。结果 TGF-β能诱导胃癌细胞NCI-N87发生EMT改变，促进癌细胞迁移和侵袭。进一步研究发现，TGF-β能诱导表皮生长因子（ EGF）和血管内皮生长因子（VEGF）表达上调，Dickkopf-1（DKK1）和分泌型卷曲受体蛋白1（SFRP1）表达降低，进而分别诱导PI3K/AKT和Wnt/β-连环蛋白（catenin）通路的激活。结论 TGF-β通过诱导胃癌微环境因子的改变促进胃癌NCI-N87细胞发生EMT。%Objective To investigate the microenvironment alteration in epithehiae-mesenchymce transition(EMT)metastasis in gastric carcinoma induced by transforming growth factor β(TGF-β). Methods The gastric carcinoma cell line NCI-N87 was treated with TGF-βto induce cells to undergone EMT,real-time quantitative PCR(RT-qPCR)and immunofluorescence staining were used to examine expression of EMT markers and wound-healing assays,and transwell migration and invasion assays were performed to determine the potential of cell migration and invasion. Further,alteration of cytokines in tumor microenvironment was detected using real-time quantitative PCR and ELISA. Moreover,the activity of its downstream signaling molecules was detected by Western blot. Results TGF-β induced gastric carcinoma cells NCI-N87 to undergone EMT as well as promote cell migration and invasion. Further,TGF-βinduced upregulation of epidermal growth foctor(EGF)and vascular endothelial growth facfor(VEGF)expression,as well as downregulation of Dickkopf-1(DKK1)and secreted frizzled receptor proein1(SFRP1),which activated PI3K/AKT and Wnt/β-catenin sequentially. Conclusion TGF-β promotes EMT by inducing microenvironment alteration in gastric carcinoma cell NCI-N87.
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