Comprehensive analysis of circular RNA profiling in AZD9291-resistant non-small cell lung cancer cell lines
Comprehensive analysis of circular RNA profiling in AZD9291-resistant non-small cell lung cancer cell lines
复制标题
AZD9291耐药非小细胞肺癌细胞系中环状RNA谱的综合分析
DOI：
10.1111/1759-7714.13032
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发表时间：
2019-04-01
期刊：
影响因子：
2.9
通讯作者：
Yang, Yunhai
中科院分区：
文献类型：
--
作者：
Chen, Tianxiang;Luo, Jizhuang;Yang, Yunhai
背景奥希替尼（AZD9291）是第三代EGFR酪氨酸激酶抑制剂，可有效延长EGFR突变尤其是T790M突变的非小细胞肺癌（NSCLC）患者的生存期；方法考虑到环状RNA（circRNA）在癌症中的重要作用，我们建立了AZD9291耐药的NSCLC细胞系（H1975/AZDR和HCC827/AZDR），并利用微阵列分析确定circRNA表达谱 的细胞。通过逐渐增加药物浓度来诱导H1975/AZDR和HCC827/AZDR细胞系。从 H1975、HCC827、H1975/AZDR 和 HCC827/AZDR 细胞中获得 CircRNA 微阵列表达谱，并通过定量逆转录 PCR 进行验证。对表达数据进行生物信息分析。结果成功建立H1975/AZDR和HCC827/AZDR细胞系。 H1975/AZDR和HCC827/AZDR细胞的半数抑制浓度和侵袭能力显着增强。 H1975/AZDR和HCC827/AZDR的增殖率远低于H1975和HCC827。微阵列分析鉴定出 15504 个 circRNA 在 H1975、HCC827、H1975/AZDR 和 HCC827/AZDR 细胞中差异表达。其中，7966个上调，7538个下调两倍以上。我们预测了最失调的 circRNA 中可能的 miRNA。此外，京都基因和基因组百科全书通路分析表明，最受调节的 circRNA 调节多种癌症和癌症相关通路。结论我们的结果表明，circRNA 可能在 NSCLC AZD9291 耐药中发挥作用，并且可能是基因治疗的有前途的候选分子靶点。
BackgroundOsimertinib (AZD9291), a third-generation EGFR-tyrosine kinase inhibitor, can effectively prolong survival in non-small cell lung cancer (NSCLC) patients with EGFR mutations, particularly T790M mutations; however, acquired resistance to AZD9291 is inevitable, thus exploration of the targets of resistance is urgent.MethodsConsidering the important role of circular RNAs (circRNAs) in cancers, we established AZD9291-resistant NSCLC cell lines (H1975/AZDR and HCC827/AZDR) and used microarray analysis to determine the circRNA expression profiles of the cells. The H1975/AZDR and HCC827/AZDR cell lines were induced by gradually increasing the drug concentration. CircRNA microarray expression profiles were obtained from H1975, HCC827, H1975/AZDR, and HCC827/AZDR cells and validated by quantitative reverse transcription PCR. Expression data were analyzed bioinformatically.ResultsThe H1975/AZDR and HCC827/AZDR cell lines were successfully established. The half-maximal inhibitory concentration and the invasion ability of H1975/AZDR and HCC827/AZDR cells were significantly enhanced. The proliferation rates of H1975/AZDR and HCC827/AZDR were much lower than H1975 and HCC827. Microarray analysis identified 15504 circRNAs differentially expressed in H1975, HCC827, H1975/AZDR, and HCC827/AZDR cells. Among them, 7966 were upregulated and 7538 were downregulated more than two-fold. We predicted the possible miRNAs of the top dysregulated circRNAs. Furthermore, Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that the most modulated circRNAs regulate several cancers and cancer-related pathways.ConclusionOur results reveal that circRNAs may play a role in NSCLC AZD9291 resistance and might be a promising molecular target candidate for gene therapy.