CircPDE4D在非小细胞肺癌获得性耐药中的作用及机制研究

Acquired drug resistance is the main obstacle to the treatment of non-small cell lung cancer （NSCLC）patients, but the roles and mechanisms of circRNAs in drug resistance for NSCLC are still unclear. In this project, circPDE4D was screened by RNA sequencing and low-expressed in NSCLC drug-resistant cell lines. Overexpression of circPDE4D significantly promoted apoptosis of A549/DDP cells in the functional assays. CircPDE4D was distributed in the cytoplasm and nucleus by FISH. The dual-luciferase report assays showed that miR-1270 might bind to circPDE4D and APAF-1 3’UTR. In addition, western blot showed that circPDE4D might up-regulate APAF-1 and miR-1270 might down-regulate APAF-1. Bioinformatics predicted that circPDE4D might bind to many proteins. Hence, we hypothesize that circPDE4D may be involved in drug resistance of NSCLC by competitively binding of miR-1270 to regulate APAF-1 or binding proteins. This project aims to explore the roles and mechanisms of circPDE4D in drug resistance of NSCLC by employing bioinformatics, molecular biology, in vitro experiments, in vivo experiments and clinical data. This project will provide new therapeutic targets and ideas to solve the problem of acquired drug resistance of NSCLC.

获得性耐药是非小细胞肺癌患者治疗的主要障碍，而目前circRNAs在非小细胞肺癌耐药中的作用及机制尚不清楚。本项目通过全转录组测序筛选出circPDE4D在非小细胞肺癌耐药细胞株中低表达；FISH实验显示circPDE4D分布于胞浆和胞核中；功能实验显示过表达circPDE4D显著促进A549/DDP细胞凋亡；报告基因实验显示miR-1270与circPDE4D和APAF-1结合；circPDE4D上调APAF-1，miR-1270下调APAF-1；生物信息学预测circPDE4D可与多种蛋白结合。据此我们提出假说：circPDE4D可能通过竞争性结合miR-1270调控APAF-1或结合蛋白参与非小细胞肺癌耐药。本项目拟利用生物信息学、分子生物学、体外实验、体内实验和临床数据等探讨circPDE4D在非小细胞肺癌耐药中的作用及机制。本项目将为克服非小细胞肺癌耐药难题提供新靶点。

肿瘤耐药问题严重限制了非小细胞肺癌的治疗效果。本项目通过全转录组测序技术，并综合利用生物信息学和细胞生物学实验等全面系统深入地研究非小细胞肺癌耐药的机制。本项目筛选了在非小细胞肺癌耐药细胞中差异表达的circRNAs、lncRNAs、miRNAs和mRNAs，构建非小细胞肺癌耐药的ceRNA调控网络，为研究ncRNAs在非小细胞肺癌耐药中的作用奠定扎实的基础。CircRNAs在非小细胞肺癌耐药中的作用及机制尚不清楚，本项目发现circPDE4D及其宿主基因PDE4D在非小细胞肺癌耐药中的重要作用及机制，为逆转非小细胞肺癌耐药提供了一种新的治疗策略。糖酵解代谢和肿瘤耐药息息相关，本项目筛选了与糖酵解代谢和肿瘤耐药均相关的ncRNAs，并通过后续实验验证结果，为解决非小细胞肺癌耐药难题开拓了新思路。