P-糖蛋白介导的化疗药物外排是肿瘤细胞产生多药耐药性的重要原因。而多药耐药性与肿瘤的发展、侵袭转移密切相关。如何发现具有逆转肿瘤耐药性及协同抗肿瘤发展、转移多重作用的化合物，对于改善化疗疗效至关重要。本项目利用建立的P-糖蛋白抑制剂的高通量筛选及活性验证的细胞模型，从中药提取物/单体成分中获得机制明确的P-糖蛋白抑制剂，进一步以乳腺癌敏感及耐药细胞为模型，以肿瘤相关信号通路的生物标志物为靶分子，通过P-糖蛋白抑制剂的靶向代谢组学研究，以及与肿瘤血管生成、侵袭转移相关的二维细胞共培养模型，靶向代谢物的在线微透析-液/质联用实时分析及细胞相互作用的代谢组学研究，结合它们对糖酵解途径相关限速酶的酶抑制活性筛选，明确具有抑制P-糖蛋白表达作用的中药活性成分的协同抗肿瘤发展机制，结合三维肿瘤多细胞球体浸润模型，探讨其抗肿瘤侵袭转移的能力，为具有多靶点、多途径抗肿瘤作用的新药研发提供科学依据。

That P-glycoprotein (P-gp) mediated drug efflux is the important reason which the tumor cells produce multi-drug resistance. The multidrug resistance is closely related to the development, invasion and metastasis of tumor. How to find reversal agents against tumor drug resistance, which have synergistic action with chemotherapy drugs on anti-tumor development and transfer, is very import to improve the curative effect of chemotherapy. In the project, the high-throughput screening method and verification mode of P - glycoprotein inhibitors will be established and they will be used for the screening and verification of P-gp inhibitors from Chinese medicine extracts/compounds. The mechanism of these new P-gp inhibitors will be clarified. Further, based on the tumor cell models, targeting metabonomics method will be used to investigate the anti-tumor mechanism of these P-gp inhibitors from Chinese medicine. The target molecules in targeting metabonomics analysis are related biomarkers with tumor signal pathways. Moreover, the activities of these P-gp inhibitors on anti-angiogenesis and invasion and metastasis of tumor will be studied using the co-culture cell model and the real-time analysis by MD-UPLC-MS/MS, and cellular metabolomics. Their inhibitory activities on related enzymes with glycolysis pathway will be screened. Based on the above research, the synergistic action mechanism of the P–gp inhibitors from Chinese medicine with chemotherapy drugs on anti-tumor will be clarified. Combined with the development of three-dimensional cell model, the anti-invasion and anti-transfer abilities will be investigated. These results will provide the scientific basis for research and development of new anti-tumor drugs with multiple targets and multiple ways.