Zhou Joshua Jiawei, Huang Yuanshen, Zhang Xue, Cheng Yabin, Tang Liren, Ma Xiaodong
Department of Anesthesiology, Pharmacology, and Therapeutics, Faculty of Medicine, University of British Columbia, Vancouver, BC, Canada.
Welichem Biotech Inc., Burnaby, BC, Canada.
Oncotarget. 2017 Sep 27;8(62):105081-105092. doi: 10.18632/oncotarget.21352. eCollection 2017 Dec 1.
EYA1 is a DNA repair enzyme that is induced after DNA damage and is upregulated in melanoma. However, its role in pathogenesis and therapeutic targeting of melanoma is unknown. Our objectives are (1) to study the relationship between EYA1 expression levels and melanoma patients' clinical pathologic parameters including survival; (2) to investigate its impact on cultured melanoma cells ; and (3) to evaluate EYA1 inhibitors' potential as a treatment of melanoma. Melanoma tissue microarrays were used to assess EYA1 protein expression in 326 melanoma tissues, and to correlate the expression with patients' clinical pathological parameters. In addition, retroviral ShRNA vectors were used to silence expression of EYA1 in A375 melanoma cells, and the resultant cells examined for changes in growth, DNA synthesis, and tumor formation . Lastly, melanoma cells were treated with benzbromarone with or without the BRAF inhibitor vemurafenib. Our results showed that EYA1 protein is low in benign nevi, but is significantly up-regulated in melanoma , and remains high in invasive and metastatic melanoma. In addition, silencing of EYA1 gene expression resulted in decreased proliferation and colony formation. These were associated with decreased cyclin D1 and increased phosphorylated histone protein γH2AX. Finally, treatment with benzbromarone, a specific inhibitor of EYA1, caused significant inhibition of melanoma cell proliferation, and increased sensitivity to the BRAF inhibitor vemurafenib. In conclusion, EYA1 gene is a pathogenic driver in melanoma pathogenesis. Targeting EYA1 may be a valuable strategy for treatment of melanoma.
EYA1是一种DNA修复酶，在DNA损伤后被诱导产生，且在黑色素瘤中表达上调。然而，其在黑色素瘤发病机制及治疗靶点方面的作用尚不清楚。我们的目标是：（1）研究EYA1表达水平与黑色素瘤患者临床病理参数（包括生存率）之间的关系；（2）研究其对培养的黑色素瘤细胞的影响；（3）评估EYA1抑制剂作为黑色素瘤治疗方法的潜力。黑色素瘤组织芯片用于评估326例黑色素瘤组织中EYA1蛋白的表达，并将该表达与患者的临床病理参数相关联。此外，利用逆转录病毒ShRNA载体沉默A375黑色素瘤细胞中EYA1的表达，并检测所得细胞在生长、DNA合成及肿瘤形成方面的变化。最后，用苯溴马隆处理黑色素瘤细胞，同时使用或不使用BRAF抑制剂维莫非尼。我们的结果显示，EYA1蛋白在良性痣中表达较低，但在黑色素瘤中显著上调，且在侵袭性和转移性黑色素瘤中仍保持高表达。此外，EYA1基因表达的沉默导致细胞增殖和集落形成减少。这些变化与细胞周期蛋白D1减少及磷酸化组蛋白γH2AX增加有关。最后，用EYA1的特异性抑制剂苯溴马隆处理可显著抑制黑色素瘤细胞增殖，并增加对BRAF抑制剂维莫非尼的敏感性。总之，EYA1基因是黑色素瘤发病机制中的致病驱动因素。靶向EYA1可能是治疗黑色素瘤的一种有价值的策略。
