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- 2018
两种细胞培养方法探究wnt/β-catenin信号通路对EMMPRIN/MMPs的影响
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Abstract:
摘要 目的:用牙龈成纤维细胞(human gingival fibroblast,HGFs)单独培养、HGFs与人永生口腔上皮细胞(human immortalized oral epithelial cell,HIOEC)直接共培养模型探究wnt/β-catenin信号通路对基质金属蛋白酶诱导因子(EMMPRIN)、明胶酶(MMP-2、MMP-9)表达的影响。方法:细胞免疫荧光检测β-catenin的表达。使用wnt/β-catenin信号通路的抑制剂Dickkopf-1(DKK-1)和激活剂wnt3a分别作用于HGFs和共培养模型。蛋白印记法、实时荧光定量PCR、明胶酶谱法检测细胞内和培养基上清中相关指标的表达。结果:β-catenin在HIOEC中表达较HGFs多。当wnt/β-catenin信号通路被抑制时,EMMPRIN、MMP-2、MMP-9的表达增加(P＜0.05),反之则减少(P＜0.05)。共培养模型较HGFs单独培养对DKK-1和wnt3a的反应更敏感。结论:共培养模型是本实验较好的实验方法。激活wnt/β-catenin信号通路可有效抑制EMMPRIN、MMP-2、MMP-9的表达
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