Department of Anatomical and Cellular Pathology, State Key Laboratory of Translational Oncology, Sir Y.K. Pao Cancer Center, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong, China.
Department of Biomedical Sciences and Center of Nasopharyngeal Carcinoma Research, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, China.
J Cell Mol Med. 2020 Nov;24(22):13523-13535. doi: 10.1111/jcmm.16007. Epub 2020 Oct 19.
Nasopharyngeal carcinoma (NPC) is an Epstein-Barr virus (EBV)-associated epithelial malignancy. The high expression of BART-miRNAs (miR-BARTs) during latent EBV infection in NPC strongly supports their pathological importance in cancer progression. Recently, we found that several BART-miRNAs work co-operatively to modulate the DNA damage response (DDR) by reducing Ataxia-telangiectasia-mutated (ATM) activity. In this study, we further investigated the role of miR-BARTs on DDR. The immunohistochemical study showed that the DNA repair gene, BRCA1, is consistently down-regulated in primary NPCs. Using computer prediction programs and a series of reporter assays, we subsequently identified the negative regulatory role of BART2-3p, BART12, BART17-5p and BART19-3p in BRCA1 expression. The ectopic expression of these four miR-BARTs suppressed endogenous BRCA1 expression in EBV-negative epithelial cell lines, whereas BRCA1 expression was enhanced by repressing endogenous miR-BARTs activities in C666-1 cells. More importantly, suppressing BRCA1 expression in nasopharyngeal epithelial cell lines using miR-BART17-5p and miR-BART19-3p mimics reduced the DNA repair capability and increased the cell sensitivity to the DNA-damaging chemotherapeutic drugs, cisplatin and doxorubicin. Our findings suggest that miR-BARTs play a novel role in DDR and may facilitate the development of effective NPC therapies.
鼻咽癌(NPC)是一种 Epstein-Barr 病毒(EBV)相关的上皮恶性肿瘤。在 NPC 中潜伏 EBV 感染时 BART-miRNAs(miR-BARTs)的高表达强烈支持它们在癌症进展中的病理重要性。最近,我们发现几种 BART-miRNAs 协同作用,通过降低共济失调毛细血管扩张突变(ATM)活性来调节 DNA 损伤反应(DDR)。在这项研究中,我们进一步研究了 miR-BARTs 在 DDR 中的作用。免疫组织化学研究表明,BRCA1 基因在原发性 NPC 中持续下调。我们使用计算机预测程序和一系列报告基因实验,随后确定了 BART2-3p、BART12、BART17-5p 和 BART19-3p 在 BRCA1 表达中的负调控作用。这四个 miR-BARTs 的异位表达抑制了 EBV 阴性上皮细胞系中的内源性 BRCA1 表达,而在 C666-1 细胞中抑制内源性 miR-BARTs 活性则增强了 BRCA1 表达。更重要的是,使用 miR-BART17-5p 和 miR-BART19-3p 模拟物抑制鼻咽上皮细胞系中的 BRCA1 表达降低了 DNA 修复能力,并增加了细胞对 DNA 损伤化学治疗药物顺铂和阿霉素的敏感性。我们的研究结果表明,miR-BARTs 在 DDR 中发挥新的作用,并可能促进有效的 NPC 治疗方法的发展。