PHP14通过与Vimentin相互作用影响TGF-β诱导的肝细胞AML-12上皮-间质转化

PHP14 Plays a Role in Epithelial-Mesenchymal Transition of AML-12 Cell Through Interaction with Vimentin

目的:探讨PHP14在上皮-间质转化中的作用及其可能作用机制。方法:建立了研究上皮-间质转化的细胞模型小鼠肝细胞AML-12。在AML-12中过表达PHP14,通过光学显微镜进行形态学观察,应用实时荧光定量PCR和蛋白质印迹检测AML-12上皮-间质转化中相关标志物的表达情况。通过免疫共沉淀检测PHP14与Vimentin的相互作用情况。结果:小鼠肝细胞AML-12可作为研究上皮-间质转化的较好细胞模型。在AML-12中过表达PHP14可促进AML-12细胞的上皮-间质转化,并增强其运动能力。过表达PHP14可上调Vimentin的蛋白质水平,而对其他上皮-间质转化相关标志物表达没有影响。进一步的研究发现,PHP14可与Vimentin相互作用。结论:PHP14通过与Vimentin相互作用,稳定Vimentin的蛋白质水平,从而参与并调节上皮-间质转化。

Objective:To investigate the role of PHP14 in epithelial-mesenchymal transition and its possible mechanism.Methods:A cell model was established to study epithelial-mesenchymal transition in mouse hepatocytes AML-12.Overexpression of PHP14 in AML-12.Morphological observation was carried out by light microscope.The expression of related markers in epithelial-mesenchymal transition of AML-12 was detected by real-time fluorescent quantitative PCR and Western blot.The interaction between PHP14 and Vimentin was detected by immunoprecipitation.Results:Mouse hepatocyte AML-12 can be used as a good cell model to study epithelial-mesenchymal transition.Overexpression of PHP14 in AML-12 can promote epithelial-mesenchymal transition and enhance the motility of AML-12 cells.Overexpression of PHP14 upregulated Vimentin,but had no effect on the expression of other epithelial-mesenchymal transition related markers.Further studies showed that PHP14 could interact with Vimentin.Conclusion:PHP14 can stabilize Vimentin protein level and participate in and regulate epithelial-mesenchymal transition,by interacting with Vimentin.