He Wei, Pang Lina, Gong Shuai, Wang Xin, Hou Lixia
Department of Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China.
Department of Radiotherapy, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China.
J Cancer. 2020 Jun 6;11(16):4801-4809. doi: 10.7150/jca.47495. eCollection 2020.
Nei endonuclease VIII-like 2 (NEIL2) is a gene encoding DNA repair enzyme, which is involved in the base excision repair (BER) pathway in mammalian cells. Cisplatin is a common cytotoxic anti-tumor agent in clinic by destroying normal structure of DNA and inducing cell apoptosis. However, how NEIL2 affects the sensitivity of NSCLC to cisplatin is still unclear. The clinical data from 206 patients diagnosed pathologically were collected. The DNA sequencing of gene 3'UTR and the PFS curve of NSCLC patients receiving cisplatin-based chemotherapy were performed. Western blot analysis and immunohistochemistry were used to detect NEIL2 protein expression. Human NSCLC cell lines A549 and H1299 were cultured and evaluated for cell viability. RT-PCR was performed for quantitative detection of miR-548a. 3'UTR reporter plasmid was constructed and luciferase reporter assay was used to verify the target gene regulated by miR-548a. In this study, we found that the gene had the polymorphism (T/C) in rs8191670 and it is associated with the PFS of advanced NSCLC patients. MiR-548a targets 3'UTR to suppress its expression. Upregulation of NEIL2 expression or downregulation of miR-548a could reduce the sensitivity of NSCLC cells to cisplatin. Our results demonstrated that NEIL2 gene rs8191670 polymorphism affects the PFS of advanced NSCLC patients, and the underlying molecular mechanisms may be that miR-548a can regulate NEIL2 expression by binding to its 3'UTR seed region containing rs8191670.
内核酸酶VIII样蛋白2（NEIL2）是一种编码DNA修复酶的基因，它参与哺乳动物细胞中的碱基切除修复（BER）途径。顺铂是临床上一种常见的细胞毒性抗肿瘤药物，可破坏DNA的正常结构并诱导细胞凋亡。然而，NEIL2如何影响非小细胞肺癌（NSCLC）对顺铂的敏感性仍不清楚。收集了206例经病理诊断患者的临床数据。对基因3'UTR进行DNA测序，并绘制接受顺铂化疗的NSCLC患者的无进展生存期（PFS）曲线。采用蛋白质免疫印迹分析和免疫组织化学检测NEIL2蛋白表达。培养人NSCLC细胞系A549和H1299，并评估细胞活力。进行逆转录聚合酶链反应（RT-PCR）定量检测miR-548a。构建3'UTR报告质粒，并使用荧光素酶报告基因检测法验证受miR-548a调控的靶基因。在本研究中，我们发现该基因在rs8191670位点存在多态性（T/C），且与晚期NSCLC患者的PFS相关。miR-548a靶向3'UTR以抑制其表达。上调NEIL2表达或下调miR-548a可降低NSCLC细胞对顺铂的敏感性。我们的结果表明，NEIL2基因rs8191670多态性影响晚期NSCLC患者 的PFS，其潜在分子机制可能是miR-548a可通过与包含rs8191670的3'UTR种子区域结合来调节NEIL2表达。
