NCI-60肿瘤细胞系转录因子蛋白质组学分析及耐药转录因子ESRRA的机制研究

Transcription factors(TF) play an important role in tumor development and anti-tumor drug therapy. In the previous study, we used a synthetic DNA containing a concatenated tandem array of the consensus TF response elements (catTFRE) as an affinity reagent and combined mass spectrometry-based proteomics to construct a transcription factor proteomic map of NCI-60 cell line. Integrating proteomics data with anti-tumor drug efficacy, we found nuclear receptor ESRRA was associated with platinum drug resistance. In the data set, a total of 849 TFs was identified, with an average of 380 per cell line. Tumor cells still retain their tissue and organ source characteristics at the transcription factor level, which partly lost at transcriptome or proteome level. Knockdown of ESRRA in HT29 cell line significantly enhanced the pharmacodynamic effect of Carboplatin. However, the mechanism by which nuclear receptor ESRRA causes platinum resistance in tumor cells remains unclear. In this study, we intend to use modern molecular biology methods such as mass spectrometry, CRISPR and ChIP-seq to explore the following questions. We aim to identify co-regulatory factors interacting with ESRRA and their downstream target genes, which may explain the mechanism of drug resistance they regulated. We will also demonstrate their clinical significance in colon cancer. This study will provide a basis for understanding the characteristics of different tumor cells at the transcription factor level, find new transcription factors related to the efficacy of anticancer drugs, and guide the combination therapy of tumors.

转录因子在肿瘤发生发展及抗肿瘤药物治疗中具有重要的作用。前期研究中，我们通过串联多拷贝双链DNA结合元件(catTFRE)富集转录因子，联用基于质谱的蛋白质组学技术，构建了NCI-60细胞系转录因子蛋白质组学图谱，并筛选出与铂类药物耐药相关的核受体ESRRA。研究发现：①共鉴定到转录因子849个，平均每种细胞系380个；②肿瘤细胞在转录因子层面仍保留其组织器官来源特征；③在HT29细胞系中，敲低ESRRA能显著提高卡铂药效。但是ESRRA产生耐药性的机制仍不清楚。本研究拟利用质谱分析、CRISPR、ChIP-seq等现代分子生物学方法，旨在：①鉴定与ESRRA作用的共调节因子；②鉴定ESRRA调控的下游靶基因，明确其产生耐药性的机理；③ESRRA在结直肠癌组织中的表达临床意义。本研究将为从转录因子层面认识不同肿瘤细胞特征、寻找与抗癌药物药效相关转录因子和指导肿瘤联合用药提供依据。