养正消积胶囊逆转非小细胞肺癌吉非替尼耐药的作用及分子机制研究

Currently, molecular targeted therapy is an important therapeutic strategy for advanced non-small cell lung cancer (NSCLC). Epidermal growth factor receptor -tyrosine kinase inhibitor ((EGFR-TKI) has become the standard targeted therapy for lung cancer treatment, however, most patients will develop drug resistance after a period of time. It is emergent to solve the challenge of the acquired drug-resistance of EGFR-TKI in NSCLC treatment. .Previous data have shown that the unfolded protein response (UPR), mediated by endoplasmic reticulum molecular chaperone prolyl4-hydroxylase beta polypeptide(P4HB) might be one of the important mechanisms of EGFR-TKI resistance. In clinical, we found that Yangzhengxiaoji Capsule (YZXJ), combined with Gefitinib had better effects than Gefitinib alone. In further study, we found that the acquired drug-resistance of Gefitinib in lung adenocarcinoma cells were reversed to a certain degree by YZXJ, and at the same time, the expression of P4HB was down-regulated. Based on reports in the literatures and our preliminary studies, we hypothesized “YZXJ might reverse the acquired drug-resistance of Gefitinib, which might be via the regulation of UPR mechanisms mediated by molecular chaperone P4HB”..To confirm this hypothesis, Gefitinib sensitive or resistant lung cancer cell lines and the tumor xenograft mouse models will be established. We also built P4HB recombinant lentivirus by overexpression and RNAi. Experiment methods such as, flow cytometry, immunofluorescence, MTT assay, immunohistochemistry staining, Real-time PCR, Western blot and many other experimental techniques will be used to explore the effects of YZXJ on the acquired drug-resistance of Gefitinib in vitro and in vivo, as well as to investigate the molecular mechanisms. .The study is expected to elucidate the effects and mechanisms of YZXJ, and thus to provide new ideas and theoretical basis for the treatment of NSCLC.

分子靶向治疗是晚期非小细胞肺癌（NSCLC）的重要治疗策略之一，但EGFR-TKIs获得性耐药问题亟待解决，分子伴侣P4HB介导的内质网应激反应可能是导致耐药的重要机制。我们研究发现，养正消积胶囊与吉非替尼联用有较好临床疗效；干预肺癌耐药细胞后可在一定程度上逆转耐药，同时P4HB表达下调，但其具体机制尚不明确。基于文献报道和前期研究，提出养正消积胶囊可能通过调控P4HB介导的内质网应激反应逆转肺癌吉非替尼耐药的假说。为验证此假说，拟分别构建吉非替尼敏感及耐药细胞株、敏感及耐药肿瘤异种移植裸鼠模型，应用基因过表达、干扰手段上调或下调P4HB表达，给予养正消积和/或吉非替尼干预，应用组织病理学、免疫荧光染色、流式细胞术、RT-PCR、Western blot等技术，在体内外研究中探讨养正消积胶囊逆转吉非替尼耐药的作用及机制。本研究将为进一步阐明肺癌吉非替尼耐药的机制及防治策略提供新思路和依据。

晚期非小细胞肺癌（NSCLC）的治疗中，分子靶向治疗是重要治疗策略之一，但EGFR-TKIs获得性耐药问题是临床常见的难点。分子伴侣P4HB介导的内质网应激反应可能是导致耐药的重要机制。本课题组前期研究发现，养正消积胶囊与吉非替尼联用有较好临床疗效；干预肺癌耐药细胞后可在一定程度上逆转耐药，同时P4HB表达下调，但其具体机制尚不明确。本研究分为体内研究和体外研究，分别构建吉非替尼敏感及耐药细胞株、敏感及耐药肿瘤异种移植裸鼠模型，应用基因过表达、干扰手段上调或下调P4HB表达，给予养正消积和/或吉非替尼干预，应用组织病理学、 疫荧光染色、流式细胞术、RT-PCR、Western blot等多种技术，在体内外研究中探讨养正消积胶囊 逆转吉非替尼耐药的作用及机制。研究发现，养正消积胶囊能逆转肺癌吉非替尼耐药，具体机制可能与调控P4HB介导的内质网应激反应有关。