三叶青黄酮类成分调控节律基因-糖酵解途径逆转NSCLC获得性EGFR-TKIs耐药的机制研究

EGFR-TKIs是治疗EGFR敏感突变型晚期非小细胞肺癌(NSCLC)的一线靶向药物，但不可避免的获得性耐药已成为其应用瓶颈。三叶青为临床常用抗肿瘤畲药，有清热解毒、化瘀止痛等功效。申请人前期研究发现三叶青黄酮类成分具有逆转NSCLC获得性EGFR-TKIs耐药；下调糖酵解增强顺铂抗NSCLC活性；选择性调控NSCLC细胞节律基因表达的作用。我们推测三叶青黄酮类成分可能通过抑制昼夜节律基因介导的糖酵解发挥逆转EGFR-TKIs耐药作用。本项目通过体外构建EGFR-TKIs耐药细胞株模型和体内裸鼠皮下成瘤模型，观察三叶青黄酮类成分对节律基因介导的糖酵解过程及EGFR-TKIs耐药的影响，阐明其逆转EGFR-TKIs耐药的药效物质基础，揭示其调控糖酵解昼夜节律逆转EGFR-TKIs耐药的作用靶点及其机制，并从时辰疗法角度优化三叶青黄酮类成分联合EGFR-TKIs治疗NSCLC的给药方案。

EGFR-TKIs are the first-line molecular targeted drugs for the treatment of advanced non-small cell lung cancer (NSCLC) patients with EGFR-sensitive mutations, but inevitable acquired resistance has seriously limited their clinical application. Tetrastigma hemsleyanum is a commonly used clinical anti-tumor She medicine as heat-clearing and detoxifying, resolving blood stasis and relieving pain. The applicant's previous study found that flavonoids from Tetrastigma hemsleyanum have the effect of reversing acquired EGFR-TKIs resistance in NSCLC. In addition, flavonoids from Tetrastigma hemsleyanum down-regulate glycolysis then enhance the antitumor activity of DDP against NSCLC. More importantly, flavonoids from Tetrastigma hemsleyanum can selectively regulate the expression of circadian rhythm genes in NSCLC cells. We hypothesize that flavonoids from Tetrastigma hemsleyanum may reverse acquired EGFR-TKIs resistance via suppressing circadian gene-mediated glycolysis. In this project, we will constructe EGFR-TKIs resistant NSCLC cell line model in vitro and subcutaneous tumorigenesis model in nude mice in vivo to observe the effects of flavonoids from Tetrastigma hemsleyanum on the circadian genes-mediated glycolysis process and EGFR-TKIs resistance. Then, reveal the pharmacodynamic material, drug targets and mechanism of flavonoids from Tetrastigma hemsleyanum on acquired EGFR-TKIs resistance of NSCLC via suppressing circadian gene-mediated glycolysis. Finally, this project will provide a basis for the chronotherapy of flavonoids from Tetrastigma hemsleyanum combined with EGFR-TKIs in the treatment of NSCLC.