雷公藤甲素逆转肺腺癌紫杉醇耐药作用特征及靶点网络构建

Non-small cell lung cancer (NSCLC) is one of the highest morbidity and mortality malignant tumor but clinical curative effect is low. Taxol is the first-line chemotherapy drugs in NSCLC treatment, but its treatment efficiency is low caused by drug resistance. Therefore, it’s count for much to find effective ways to reverse or prevent resistance in NSCLC treatment. Our previous studies have found that tripterygium wilfordii element (TPL) has the effect of reversing drug resistance in lung adenocarcinoma paclitaxel resistant cells A549 / Taxol. This effect may be related to downregulate the expression of target gene ABCB1 and TUBB3, thus disturb microtubules network stability. Our working hypothesis is put forward that TPL play a role of multi-channel interaction and network control in reversing Taxol resistant multi-channel interaction and network control in lung adenocarcinoma taxol resistance by regulating the transmembrane efflux transporters, stabilize tubulin, inhibition of apoptosis protein expression abnormal implementation. We explore the regulatory network mechanism of TPL reversing taxol drug resistance by using network pharmacology. We verify the regulation and control mechanism and the relevant network signaling pathways of TPL response to lung adenocarcinoma taxol resistance by using pharmacology and molecular biology technology in resistant cells and resistant animal model. We build the reversing drug-resistance characteristic and target network of Triptolide response to lung adenocarcinoma of taxol resistance to provide a scientific basis in reversing tumor chemotherapy drug resistance areas，realize the equal emphasis on Chinese and western medicine.

非小细胞肺癌（non-small cell lung cancer, NSCLC）发病率高、临床疗效不高、死亡率高，紫杉醇是NSCLC一线化疗药因耐药而致疗效低下；因此，逆转或预防耐药成为NSCLC的治疗瓶颈与难题。我们前期研究发现雷公藤甲素（TPL）对肺腺癌紫杉醇耐药细胞A549/Taxol有逆转作用，其机制可能与下调靶点基因ABCB1和TUBB3表达进而干扰微管网络稳定相关。据此，提出“TPL通过调控跨膜外排转运蛋白、稳定微管蛋白不发生突变、抑制凋亡蛋白表达异常实现对紫杉醇耐药逆转的多途径交互作用和网络调控”的工作假说，运用网络药理学方法构建TPL逆转肿瘤耐药互作网络，运用药理学、分子生物学技术在耐药细胞、耐药动物模型中验证TPL逆转紫杉醇耐药的网络调控机制和相关信号通路，构建TPL逆转肺腺癌紫杉醇耐药作用特征及靶点网络，为肿瘤化疗药物耐药领域实现“中西医并重”提供科学依据。

紫杉醇在治疗非小细胞肺癌（non-small cell lung cancer, NSCLC）中常因耐药而导致效率低下，因此逆转或预防耐药成为治疗NSCLC的关键问题之一。雷公藤甲素（Triptolide）是中药雷公藤的主要有效成分，具有抗炎、镇痛、抗肿瘤及免疫调节等作用。本项目利用前期建立的A549和H460非小细胞肺癌紫杉醇耐药株，采用二代高通量测序技术，绘制了耐紫杉醇细胞株和敏感株之间的差异表达谱，以及雷公藤甲素处理紫杉醇耐药株后的表达谱，为研究雷公藤甲素逆转紫杉醇耐药肺癌细胞的特征及靶点网络构建打下良好基础。项目通过GO、KEGG和GSEA富集分析得到了耐药相关通路，并进一步通过实验发现雷公藤甲素逆转紫杉醇耐药的2种可能机制：1、因EMT被认为是肿瘤耐药的一个新的重要机制，本项目发现雷公藤甲素可通过经典的Wnt/β-catenin通路抑制耐紫杉醇A549细胞内皮-间充质转化；2、雷公藤甲素通过阻断经典SonicHedgehog（SHH）信号通路恢复了对NSCLC紫杉醇耐药细胞的敏感性。本项目还发现雷公藤甲素通过增加HSP70的转录活性进而促进MDM2泛素连接酶介导的TP53R175H /Y220C突变蛋白的降解，这为雷公藤甲素治疗表达P53获得性功能突变蛋白的非小细胞肺癌患者奠定理论基础。此外，项目还对雷公藤甲素调控耐紫杉醇肺癌肿瘤的肿瘤微环境进行了初步探索。