Huang Dabing, Bian Geng, Pan Yueyin, Han Xinghua, Sun Yubei, Wang Yong, Shen Guodong, Cheng Min, Fang Xiang, Hu Shilian
Shandong University School of Medicine, Jinan, 250012 China.
Department of Geriatrics, Anhui Provincial Hospital, Anhui Medical University, Hefei, 230031 Anhui China.
Cancer Cell Int. 2017 Mar 1;17:32. doi: 10.1186/s12935-017-0389-7. eCollection 2017.
MicroRNAs (miRNAs) was reported to be involved in cancer radio-resistance, which remains a major obstacle for effective cancer therapy.
The differently expressed miRNAs were detected by RNA-seq experiment in nasopharyngeal cancer (NPC) cells. MiR-20a-5p was selected as our target, which was subject to finding its target gene Rab27B via bioinformatics analysis. The qRT-PCR, western blot and the luciferase reporter assays were performed to confirm Rab27B as the target of miR-20a-5p. In addition, the roles of miR-20a-5p in NPC radio-resistance were detected by transfection of either miR-20a-5p-mimic or miR-20a-5p-antagomiR. The involvement of Rab27B with NPC radio-resistance was also detected by the experiments with siRNA-mediated repression of Rab27B or over-expression of GFP-Rab27B. Wound healing and invasion assays were performed to detect the roles of both miR-20a-5p and Rab27B.
MiR-20a-5p promotes NPC radio-resistance. We identified that its target gene Rab27B negatively correlates with miR-20a-5p-mediated NPC radio-resistance by systematic studies of a radio-sensitive (CNE-2) and resistant (CNE-1) NPC cell lines. Repression of Rab27B by siRNA suppresses cell apoptosis and passivates CNE-2 cells, whereas over-expression of Rab27B triggered cell apoptosis and sensitizes CNE-1 cells.
MiR-20a-5p and its target gene Rab27B might be involved in the NPC radio-resistance. Thus the key players and regulators involved in this pathway might be the potential targets for developing effective therapeutic strategies against NPC.
据报道,微小RNA(miRNA)与癌症放射抗性有关,而癌症放射抗性仍然是有效癌症治疗的主要障碍。
通过RNA测序实验检测鼻咽癌(NPC)细胞中差异表达的miRNA。选择miR-20a-5p作为研究对象,通过生物信息学分析寻找其靶基因Rab27B。进行qRT-PCR、蛋白质免疫印迹和荧光素酶报告基因检测以确认Rab27B是miR-20a-5p的靶基因。此外,通过转染miR-20a-5p模拟物或miR-20a-5p拮抗剂来检测miR-20a-5p在NPC放射抗性中的作用。通过siRNA介导的Rab27B抑制或GFP-Rab27B过表达实验也检测了Rab27B与NPC放射抗性的关系。进行伤口愈合和侵袭实验以检测miR-20a-5p和Rab27B的作用。
miR-20a-5p促进NPC放射抗性。通过对放射敏感(CNE-2)和抗性(CNE-1)NPC细胞系的系统研究,我们确定其靶基因Rab27B与miR-20a-5p介导的NPC放射抗性呈负相关。siRNA抑制Rab27B可抑制细胞凋亡并使CNE-2细胞失活,而Rab27B的过表达则触发细胞凋亡并使CNE-1细胞敏感。
miR-20a-5p及其靶基因Rab27B可能参与NPC放射抗性。因此,该途径中涉及的关键参与者和调节因子可能是开发针对NPC的有效治疗策略的潜在靶点。