长链非编码RNA NEAT1通过诱导BIN1选择性剪接促进肺腺癌EGFR-TKI耐药机制研究

EGFR-TKI耐药是影响肺腺癌靶向治疗效果最主要原因，目前仍有部分机制不明。促凋亡信号分子失活是EGFR-TKI耐药重要标志，其调控因素复杂。lncRNA在包括选择性剪接调控在内的多种肿瘤相关事件中发挥关键作用。我们前期研究发现，促凋亡分子桥接整合因子1（BIN1）在肺腺癌中低表达并诱导EGFR-TKI耐药，同时异常选择性剪接是导致其低表达主要原因；通过基因测序和overlap分析发现lncRNA NEAT1在肺腺癌中同时与BIN1异常选择性剪接和EGFR-TKI耐药有关。由此提出假说：NEAT1通过诱导BIN1发生异常选择性剪接促进EGFR-TKI耐药。本项目将从临床样本、细胞实验、动物模型多层次，利用RIP、剪接模式分析和TUNEL等技术深入探讨其分子调控机制。系统性绘制NEAT1参与肺腺癌EGFR-TKI耐药过程的分子调控网络，为逆转EGFR-TKI耐药治疗提供新的理论依据。

EGFR-TKI has become one of the most common and efficient option in treating lung adenocarcinoma. However, resistance severely restricted the treatment efficacy of EGFR-TKIs. The frequent mechanisms of epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) resistance include T790M mutation, c-MET amplification, PIK3CA mutation, pathological phenotype transition, MAPK pathway continuous activation, and epithelial mesenchymal transition, etc. Unfortunately, the mechanisms of about 30% cases of EGFR-TKI resistance remain elusive. Inactivation of pro-apoptotic protein is one of the hallmarkers of EGFR-TKI resistance. Due to its complexity, the mechanisms of pro-apoptotic protein anergy is still little understood. Long non-coding RNA (lncRNA) is a kind of non-encoding RNA transcripts > 200 nucleotides in length and recently identified as crucial regulators of gene expression in cancer. Our preliminary study revealed that the low expression of pro-apoptotic protein bridging integrator-1 (BIN1) is correlated with EGFR-TKI resistance. At the same time, we found that aberrant alternative splicing accounted for this low expression. Aiming at elucidating the related factors, we assayed the lncRNA expression profile with sequencing chips, then we used overlap analysis and found that lncRNA NEAT1 was correlated with BIN1 aberrant alternative splicing and EGFR-TKI resistance simultaneously. Taken above, we put forward a hypothesis that NEAT1 could promote lung adenocarcinoma EGFR-TKI resistance through inducing aberrant BIN1 alternative splicing. In this project, we plan to use molecular biological methods including RIP, splicing analysis, and TUNEL, etc. to explore the regulating mechanisms from clinical sample, cell experiment, and in vitro model aspects. Thereout, to systematically describe the molecular regulation network of NEAT1 in EGFR-TKI resistance and provide novel theoretical foundation to reverse EGFR-TKI resistance.