LncRNAs在非小细胞肺癌EGFR-TKIs耐药中的作用及分子机制

The mechanisms of EGFR-TKIs resistance have not been yet fully elucidated. Our research group in previous studies found that the activation of PI3K/AKT signaling and BIM downregulation was associated with EGFR-TKIs resistance. Long non-coding RNAs (lncRNAs) are closely related with cancers by involving in the regulation of cell apoptosis, proliferation and other functions.In our pre-experiment we analysized lncRNAs expression profiles using lncRNAs microarray in both EGFR-TKIs sensitive and resistant cell lines, and found over 4000 significantly different lncRNAs, and captured four target lncRNAs by bioinformatic analysis that were associated with cell apoptosis and proliferation: UCA1, BC200, BC087858 and HOTAIR. Moreover, we validated these lncRNAs using qRT-PCR in NSCLC cell lines. We speculated that lncRNAs may be involved in the EGFR-TKIs resistance, but the specific regulatory mechanisms need to be further clarified. Therefore, we will explore the role and molecular mechanisms of lncRNAs on EGFR-TKIs resistance by gene transfection, western blot and other methods respectively in vitro and vivo. In addition, the predictive value of them to EGFR-TKIs resistance will be evaluated by clinical samples.

EGFR-TKIs耐药机制仍未完全阐明。课题组前期研究发现PI3K/AKT信号活化和BIM表达下调与EGFR-TKIs耐药相关。LncRNAs与肿瘤关系密切，参与细胞凋亡、增殖等功能调控。课题组预实验采用LncRNAs芯片分析了EGFR-TKIs敏感株和耐药株的lncRNAs表达谱差异，发现了4000多个差异明显的lncRNAs，通过生物信息学分析捕获了四个与凋亡和增殖相关的目标lncRNAs：UCA1、BC200、BC087858和HOTAIR，并用定量RT-PCR在细胞株中进行验证。我们推测这些LncRNAs可能参与了EGFR-TKIs耐药，但具体调控机制有待阐明。因此，本课题拟采用基因转染、Westernblot等方法分别在细胞和动物水平探讨目的LncRNAs在EGFR-TKIs耐药中的作用及分子机制，并用临床标本加以验证，评估它们能否成为预测或干预EGFR-TKIs耐药的分子靶标。

本项目探讨了lnRNAs UCA1、HOTAIR、BC087858在EGFR-TKIs耐药中的作用和分子机制。通过细胞水平和动物水平的实验课题组研究发现，UCA1和BC087858通过PI3K/AKT信号通路发挥耐药作用，而HOTAIR通过EMT在EGFR TKIs耐药中发挥作用。同时课题组建立了EGFR-TKIs耐药NSCLC患者的样本库，通过临床标本验证进一步证实了UCA1、HOTAIR、BC087858在EGFR-TKIs耐药中的作用，发现UCA1和BC087858在获得性耐药患者中表达明显升高，而HOTAIR在原发性和获得性耐药患者中表达明显降低。此外，课题组还探讨了H19在顺铂耐药中的作用和相关机制。而lnRNAs能否成为预测EGFR-TKI耐药有效的生物标志物仍需要进一步的研究加以证实。