ecDNA中MYC/WWP1信号轴调控与卵巢癌耐药和转移的桂枝茯苓丸干预研究

染色体外DNA（ecDNA）中MYC、WWP1、EGFR等癌基因扩增导致的肿瘤异质性是多药耐药（MDR）和上皮间质转化（EMT）的重要原因，多种信号通路参与了MDR和EMT调控。MYC/WWP1信号轴调控PTEN异常导致EGFR和PI3K/AKT通路活化，促进卵巢癌MDR和EMT，与患者总生存率下降密切相关。前期研究发现桂枝茯苓丸（GFW）可上调卵巢癌耐药细胞中PTEN表达，抑制PI3K/AKT通路活化。据此，我们提出假说：桂枝茯苓丸通过靶向ecDNA中MYC/WWP1信号轴调控PTEN，逆转卵巢癌MDR和EMT。本研究拟在卵巢癌耐药细胞和耐药性移植瘤和转移瘤小鼠模型中，采用CRISPR/Cas9、FISH、ATAC-seq/see等技术，明确其干预ecDNA中MYC/WWP1信号轴，抑制MDR和EMT的关键环节和靶点，深入探讨该方逆转卵巢癌转移和耐药的多靶点、多通路调控机制。

Tumor heterogeneity caused by the amplification of oncogenes such as MYC, WWP1, and EGFR in extrachromosomal DNA (ecDNA) has an important role in tumor multidrug resistance (MDR) and epithelial-mesenchymal transformation (EMT). Multi-signal pathways are involved in the regulation of MDR and EMT. Abnormality regulated PTEN by MYC/WWP1 signal axis leads to activation of EGFR and PI3K/AKT pathways, accelerating MDR and EMT of ovarian cancer (OV), and is closely related to the decline in the overall survival rate of the OV patients. Previous studies have found that Guizhi Fuling Wan (GFW) can up-regulate PTEN expression and inhibit PI3K/AKT pathway activation in ovarian cancer drug-resistant cells. Based on this, we propose a hypothesis: GFW is targeting the MYC/WWP1 signal axis in ecDNA, and then regulate PTEN and reverse MDR and EMT in OV. This study will use CRISPR/Cas9, FISH, ATAC-seq/see and other technologies, in ovarian cancer resistant cells and xenograft/metastatic mouse resistance models, to clarify GFW’s intervention effect on MYC/WWP1 signal axis in ecDNA, and then to inhibit the key links and targets related to MDR and EMT. This study is an in-depth discussion of the multi-targets and multi-pathways regulatory mechanisms by GFW reversing OV metastasis and drug resistance.