根据转录组测序技术分析介导肺癌奥希替尼耐药的信号通路，发现潜在干预靶点。

体外构建肺癌奥希替尼一线/二线治疗模式下配对的敏感/耐药细胞株，采用高通量测序技术检测耐药前后细胞转录本表达量并使用R-package CORNAS筛选差异表达基因，借助Venny在线工具分析两组配对细胞株差异表达基因的交集，运用DAVID Bioinformatics Resources数据库分别对两种耐药模式下共有差异基因和特有差异基因进行京都基因与基因组百科全书(kyoto Encyclopedia of Genes and Genomes, KEGG)信号通路和基因本体(gene ontology, GO)功能富集分析。采用STRING 12.0数据库分析关键通路富集基因相互作用关系及靶点分子，借助chiplot在线工具绘制分子相互作用网络图。

两组配对细胞株共有差异表达基因显著富集在Cell Cycle通路，一线治疗耐药特有差异基因富集在Cell Cycle和Ribosome biogenesis in eukaryotes通路，二线治疗耐药特有差异基因富集在MAPK signaling通路；GO富集分析发现共有差异表达基因参与cell division，分子功能是ATP binding，一、二线治疗耐药特有差异表达基因分别参与了rRNA processing和inflammatory response生物过程，主要分子功能分别是RNA binding和Protein binding；共筛选出5种关键靶点分子，包括CCNB1(共有)、CDC25A和NOP56(一线耐药特有)、JUN和MYC(二线耐药特有)。

奥希替尼一线或二线治疗模式下共同及特有的潜在获得性耐药机制，为克服奥希替尼耐药提供潜在的治疗靶点。

To explore the signaling pathways mediating Osimertinib resistance in lung cancer based on transcriptome sequencing technology and to identify potential targets for intervention.

In vitro, we constructed paired sensitive/resistant lung cancer cell lines under the first/second line treatment modes of Osimertinib, detected the expression of cell transcripts before and after drug resistance by high-throughput sequencing and screened the differentially expressed genes by R-package CORNAS, analyzed the intersection of differentially expressed genes between the two groups of paired cell lines with the help of Venny online tool, and using the DAVID Bioinformatics Resources database to perform kyoto Encyclopedia of Genes and Genomes(KEGG) signaling pathway and gene ontology(GO) function enrichment analysis on the common and unique differentially expressed genes in two drug-resistant modes, respectively. Finally, the STRING 12.0 database was used to analyze the key pathway enriched gene interactions and target molecules, and the molecular interaction network was mapped with the help of chiplot online tool.

The shared differentially expressed genes of the two paired cell lines were significantly enriched in the Cell Cycle pathway, the first-line treatment resistance-specific differentially expressed genes were enriched in the Cell Cycle and Ribosome biogenesis in eukaryotes pathway, and the second-line treatment resistance-specific differentially expressed genes were enriched in the MAPK signaling pathway; GO enrichment analysis revealed that shared differentially expressed genes were involved in cell division, and their molecular function was ATP binding, while first- and second-line treatment resistance-specific differentially expressed genes were involved in rRNA processing and inflammatory response, and their main molecular functions were RNA binding and Protein binding, respectively; Five key target molecules were screened, including CCNB1 (common), CDC25A and NOP56 (first-line drug resistance-specific), JUN and MYC (second-line drug resistance-specific).

The study identified potential acquired resistance mechanisms common and specific to first-or second-line treatment modalities of Osimertinib, providing potential therapeutic targets for overcoming Osimertinib resistance.