BackgroundDNA damage repair is frequently dysregulated in high grade serous ovarian cancer (HGSOC), which can lead to changes in chemosensitivity and other phenotypic differences in tumours. RFWD3, a key component of multiple DNA repair and maintenance pathways, was investigated to characterise its impact in HGSOC.MethodsRFWD3 expression and association with clinical features was assessed using in silico analysis in the TCGA HGSOC dataset, and in a further cohort of HGSOC tumours stained for RFWD3 using immunohistochemistry. RFWD3 expression was modulated in cell lines using siRNA and CRISPR/cas9 gene editing, and cells were characterised using cytotoxicity and proliferation assays, flow cytometry, and live cell microscopy.ResultsExpression of RFWD3 RNA and protein varied in HGSOCs. In cell lines, reduction of RFWD3 expression led to increased sensitivity to interstrand crosslinking (ICL) inducing agents mitomycin C and carboplatin. RFWD3 also demonstrated further functionality outside its role in DNA damage repair, with RFWD3 deficient cells displaying cell cycle dysregulation, reduced cellular proliferation and reduced migration. In tumours, low RFWD3 expression was associated with increased tumour mutational burden, and complete response to platinum chemotherapy.ConclusionRFWD3 expression varies in HGSOCs, which can lead to functional effects at both the cellular and tumour levels.

中文翻译：

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RFWD3 调节对铂类化疗的反应并促进高级别浆液性卵巢癌的癌症相关表型

背景 DNA 损伤修复在高级别浆液性卵巢癌 (HGSOC) 中经常失调，这可能导致化疗敏感性的变化和肿瘤的其他表型差异。 RFWD3 是多种 DNA 修复和维护途径的关键组成部分，我们对其进行了研究，以表征其对 HGSOC 的影响。方法使用以下方法评估 RFWD3 表达及其与临床特征的关联：计算机模拟在 TCGA HGSOC 数据集中进行分析，并在使用免疫组织化学对 RFWD3 染色的 HGSOC 肿瘤的进一步队列中进行分析。使用 siRNA 和 CRISPR/cas9 基因编辑在细胞系中调节 RFWD3 表达，并使用细胞毒性和增殖测定、流式细胞术和活细胞显微镜对细胞进行表征。结果 RFWD3 RNA 和蛋白质的表达在 HGSOC 中存在差异。在细胞系中，RFWD3 表达的减少导致对链间交联 (ICL) 诱导剂丝裂霉素 C 和卡铂的敏感性增加。 RFWD3 还展示了其 DNA 损伤修复作用之外的更多功能，RFWD3 缺陷细胞表现出细胞周期失调、细胞增殖减少和迁移减少。在肿瘤中，RFWD3 低表达与肿瘤突变负荷增加以及对铂类化疗的完全反应相关。结论 RFWD3 表达在 HGSOC 中存在差异，这可能导致细胞和肿瘤水平的功能影响。