目的:在构建三阴性乳腺癌(TNBC)的蛋白质相互作用网络基础上,探讨化痰散结方对TNBC MDAMB-231细胞增殖抑制作用以及对Hedgehog(Hh)通路转录因子Gli1表达的影响。方法:筛选在线人类孟德尔遗传数据库(OMIM)获得TNBC相关基因;使用STRING进行文本挖掘并构建TNBC蛋白质相互作用网络;将互作数据读入Cytoscape3.4.0,分别应用插件JEPETTO 1.3.1和MCODE1.4.2实现拓扑分析和聚类分析;基于DAVID数据库富集关键通路用于表征簇的生物学重要性。针对关键通路Hh及其转录因子Gli1,采用MTT法检测化痰散结方对MDAMB-231细胞的增殖抑制率,Western blot法检测Gli1蛋白的表达情况。结果:构建了由1 537个节点(蛋白质)和23 681个边(相互作用)组成的蛋白质相互作用网络,分析得出关键通路H h信号传导通路和基因G l i1。实验结果表明化痰散结方能抑制MDA-MB-231细胞的增殖并能降低Gli1蛋白的表达。结论:TNBC蛋白质相互作用网络是一个受多信号通路传导、多基因调控的复杂过程,其中H h通路及其下游转录基因G l i1很可能是关键节点,而化痰散结方对MDA-MB-231细胞增殖的抑制作用机制之一是降低Hh通路中Gli1的表达。
Objective: To discuss the effects of Huatan Sanjie Formula on expression of transcription factor Gli1 in Hedgehog signaling pathway and the inhibition of TNBC MDA-MB-231 cells proliferation based on the protein interaction network. Methods: TNBC related genes were gotten by screening the Online Mendelian Inheritance in Man(OMIM) database. The interaction data were put in Cytoscape 3.4.0 software to implement topological analysis and cluster analysis by JEPETTO 1.3.1 and MCODE 1.4.2 before constructing TNBC protein interaction network with text mining in STRING based on the essential pathway of DAVID datebase enrichment was used for biological importance of representation cluster. The expressions of protein Gli1 were detected with Western blot and the inhibition ratio of TNBC MDA-MB-231 cells proliferation was determined by MTT. Results: The protein interaction network was constructed that included 1 537 nodes(proteins) and 23 681 sides(interactions) and analyzed the Hedgehog essential pathway and gene Gli1. The research showed that Huatan Sanjie Formula could inhibit the TNBC MDA-MB-231 cells proliferation and reduce the expression of Gli1 protein. Conclusion: TNBC protein interaction network is a complex process that was mediated by multiple signaling pathways and regulated by multiple genes, and the Hedgehog signaling pathway and its downstream transcription gene Gli1 as essential point, while, one of the mechanism about inhibition of Huatan Sanjie Formula to the TNBC MDAMB-231 cells proliferation is related to reduce the expression of Gli1 protein in Hedgehog signaling pathway.
China Journal of Traditional Chinese Medicine and Pharmacy