IGF1R-RACK1-STAT3通路在苦蘵内酯P抗EGFR T790M突变非小细胞肺癌中的作用研究

项目名称： IGF1R-RACK1-STAT3通路在苦蘵内酯P抗EGFR T790M突变非小细胞肺癌中的作用研究

项目编号： No.81503297

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 戴春艳

作者单位： 浙江中医药大学

项目金额： 18万元

中文摘要： EGFR T790M继发性突变耐药NSCLC患者是现阶段治疗的难点。我们前期研究发现清热解毒类中药灯笼草的新型抗肿瘤活性成分苦蘵内酯P显著抑制T790M突变NSCLC细胞增殖和促进凋亡，其作用机制可能与其下调IGF1R近膜区Y950和STAT3的磷酸化水平有关。本研究拟通过移植瘤模型验证苦蘵内酯P对T790M突变NSCLC的抑制效应，免疫共沉淀和IGF1R Y950F突变质粒转染等方法明确苦蘵内酯P抑制IGF1R Y950，阻抑IGF1R-RACK1-STAT3复合体形成的分子机制，应用免疫印迹、免疫荧光等技术阐明苦蘵内酯P抑制IGF1R-STAT3通路活性，下调T790M突变NSCLC细胞增殖、抗凋亡相关靶基因转录表达的作用机制。通过本项目研究，揭示苦蘵内酯P抗T790M突变NSCLC的作用及机制，不仅为其今后研发和联合用药提供科学依据，而且为临床治疗T790M突变NSCLC提供新思路。

中文关键词： 苦蘵内酯P；非小细胞肺癌；胰岛素样生长因子1受体；信号转导子与转录活化子3

英文摘要： Mounting evidence showed that the major problem in NSCLC treatment is drug resistance caused by the T790M mutation in EGFR kinase. Thus, development of novel anti-tumor agents for treating NSCLC is urgently needed. Traditional Chinese herb Physalis angulata has the function of clear heat and detoxifying. Recently, our group isolated the active ingredient of Physalis angulata, physagulin P. We found that physagulin P could significantly inhibit cell proliferation and promote apoptosis by reducing the phosphorylation level of IGF1R and its downstream signaling molecule STAT3 in NSCLC cells with the EGFR T790M mutation. In this proposal, we aim to investigate the molecular mechanism underlying the therapeutic potential of physagulin P in T790M-positive NSCLC cells. Both cell model and xenograft model of T790M-positive NSCLC will be used to investigate the therapeutic efficacy of physagulin P. Co-immunoprecipitation and dominant negative mutant approach will be applied to prove the specific inhibitory role of physagulin P to IGF1R Y950, which could prevent the interaction between adaptor proteins RACK1 and STAT3. Additionally, Western blotting, immunofluorescence, and dual luciferase assays will be performed to detect the phosphorylation levels of STAT3 and its downstream targets. Our findings will not only provide the scientific evidence for the development of a next-generation IGF1R inhibitor physagulin P, but also shed new insights on improving the clinical outcomes of NSCLC patients with the EGFR T790M mutation by designing a new combined chemotherapy regimen.

英文关键词： physagulin P;non-small cell lung cancer;IGF1R;STAT3