补精解毒方调控表皮生长因子受体信号通路治疗非小细胞肺癌的分子机理研究

Bujingjiedu had good clinical efficacy on NSCLC. In early animal experiments it showed that bujingjiedu had anti-cancer activity, with a higher rate of apoptosis.Subject adopts electrophoretic mobility change experimental explorated without resistance cell line inhibition of EGFR downstream signal transduction pathways and find the best compatibility.Build acquired resistance of erlotinib cell line model, using Western Blot analysis of EGFR in cell signaling molecules of the protein changes, to study the regulation for the function of the resistance.Using normal and drug resistance lung cancer cells tumor-burdened reconstructed to establish a nude mice model and resistance model and simulating the biological behavior of lung cancer cells in vivo to study drug intervention.This study aims to clear regulation of EGFR signaling pathway in NSCLC from genetically modified and protein levels, first proposes effect of the different compatibility proportion and mechanismor for resistant and non resistant NSCLC, and supports further development of traditional Chinese medicine in the treatment of tumor.

补精解毒方（冬虫夏草、白花蛇舌草）是我科经验方，临床上治疗非小细胞肺癌疗效确切。前期动物实验表明抗肺癌活性强，具有较高的凋亡率。课题采用电泳迁移率改变实验探查补精解毒方对无耐药细胞株EGFR下游信号转导通路的抑制作用并寻找最佳配伍。构建厄洛替尼获得性耐药细胞株模型，利用Western Blot分析细胞中EGFR信号通路中信号分子蛋白的变化，从细胞水平研究其调控厄洛替尼耐药的功能。利用普通肺癌细胞及耐药肺癌细胞原位移植法建立裸鼠荷瘤模型及耐药模型，模拟体内肺癌细胞的生物学行为，研究药物的干预作用。本研究旨在从基因修饰及蛋白水平明确补精解毒方对NSCLC中EGFR信号通路的调控，首次提出对于耐药与非耐药NSCLC，补精解毒方的不同配伍比例的治疗作用与机制，进一步发展中医扶正祛邪在肿瘤治疗中的指导意义。

本研究针对补精解毒方在临床上对EGFR突变型NSCLC有较好的效果，结合药效学与有效成分富集等方法，发现在补精解毒方中熊果酸具有很强的对EGFR二次突变的非小细胞肺癌的抑制作用。使用H1975细胞建立EGFR二次突变的非小细胞肺癌细胞和动物模型，深入探索熊果酸作用机制。发现熊果酸对EGFR二次突变的非小细胞肺癌的抑制作用与其抑制CT45A2表达密切相关；进一步研究表明熊果酸能通过抑制β-catenin激活进而抑制CT45A2表达。本研究初步揭示了补精解毒方对EGFR二次突变的非小细胞肺癌抑制作用及其作用机制，发表5篇SCI论文，1篇中文核心。