基于对AKT活性的调控效应研究黄芪抑制MSCs介导肺癌细胞对厄洛替尼耐药性的生物学机制

The emergence of targeted-drug resistance is a major cause of lung cancer with poor prognosis, re-occurrence and metastasis. Mesenchymal stem cells (MSCs) can locate near tumor and secret cytokines, helping construct drug-resistance. Astragalus mongholicus has the anti-tumor efficacy. This project presumes that combination of Chinese Traditional Medicine Astragalus mongholicus and targeted-drug erlotinib contributes to reduce MSCs-mediated drug resistance of lung cacner cells, based on the “bu yang yi qi and fu zheng qu xie” theory of Chinese Traditional Medicine Astragalus mongholicus. Having found that Astragalus mongholicus has the ability to reduce activities of resistance-related molecule AKT, we intend to explore the possible way by combining Chinese with Western Medicine to reduce targeted-drug resistance of lung cancer cells. Firstly, we aim to construct targeted-drug resistance model by co-culturing EGFR-mutant lung cancer cell line HCC827 and MSCs in vitro, plus with erlotinib administration. Secondly, we intend to identify the key cytokines involving appearance of erlotinib-resistance in HCC827 by using antibody-chip technique and detect its influence on AKT signals by using Western blot analysis. Finally, we aim to explore the anti-tumor effect on HCC827 and inhibitory effct on AKT by treatment of Astragalus mongholicus and erlotinib to drug-resistance model, hoping to prevent drug-resistance of tumor cells by combination of traditional Chinese and Western medicine. We expect to explore the mechanism of MSCs-meidated drug-resistance and the way for combination Astragali with chemotherapy in the treatment of lung cancer.

耐药性是肺癌治疗预后差、易复发、易转移的主要原因。间充质干细胞（MSCs）可定位于肿瘤周围并分泌细胞因子作用于肿瘤细胞，有利于耐药性的形成。中药黄芪具有防癌抗癌的功能。本项目基于黄芪“补阳益气、扶正祛邪”理论，提出联合使用黄芪及靶向药物厄洛替尼有助于降低MSCs介导的肺癌细胞耐药的假说。本项目在观察黄芪抑制肺癌细胞耐药相关分子AKT活性的基础上，研究利用黄芪降低肺癌细胞耐受厄洛替尼的可能性；通过EGFR突变的人肺癌细胞株HCC827与MSCs进行共培养及靶向药物厄洛替尼干预，建立HCC827细胞的体外耐药模型；利用抗体芯片确定诱发HCC827细胞产生耐药性的关键细胞因子，通过蛋白印记检测该因子对癌细胞内AKT活性的影响；检测黄芪与厄洛替尼共同干预对耐药HCC827细胞的杀伤及对AKT活性的抑制作用，从而探索MSCs介导HCC827产生耐药性的机制及黄芪联合化疗降低肺癌耐药性的途径。