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EBV-miR-BART6-5p 靶向 Dicer1 对鼻咽癌细胞生物学特性及放射敏感性的影响。

Effects of Dicer1 targeted by EBV-miR-BART6-5p on biological properties and radiosensitivity of nasopharyngeal carcinoma.

机构信息

Department of Stomatology, Jingzhou Central Hospital, The Second Clinical Medical College, Yangtze University, Jingzhou, China.

Department of Otolaryngology, 549615The People's Hospital of Rizhao, Rizhao, China.

出版信息

Hum Exp Toxicol. 2021 Jun;40(6):977-993. doi: 10.1177/0960327120979020. Epub 2020 Dec 11.

Abstract

OBJECTIVE

To discuss the effects of Epstein-Barr virus (EBV)-encoded BamHI A rightward transcript (BART) microRNA (miR-BART6-5p) by targeting on biological properties and radiosensitivity of nasopharyngeal carcinoma (NPC).

METHODS

NPC patients (n = 96) treated with radiotherapy were collected from Jan 2010 to Jan 2011. Real-time quantitative PCR (qRT-PCR) and western blot were carried out to measure the expression of miR-BART6-5p and Dicer1. Dual luciferase reporter gene assay verified that miR-BART6-5p targeted . CCK8, wound-healing, Transwell and Annexin-FITC/PI were employed to evaluate the effects of Dicer1 mediated by miR-BART6-5p on biological characteristics of NPC cells. The radiosensitivity of miR-BART6-5p targeting was assessed and .

RESULTS

Increased miR-BART6-5p and decreased were discovered in NPC patients, displaying a close association with T-stage, clinical stage, as well as Pre-DNA of NPC. While elevated Dicer1 and miR-BART6-5p down-regulation in NPC patients were found after effective radiotherapy. Both miR-BART6-5p and Dicer1 were prognostic factors of NPC. Down-regulation of miR-BART6-5p could enhance Dicer1 expression and inhibit NPC cell proliferation, invasion and migration with promoted apoptosis. Clone formation assay also showed miR-BART6-5p down-regulation reduced planting efficiency (PE), which further decreased with the increased dose of irradiation. Injection with miR-BART6-5p inhibitors in nude mice after 6-Gy irradiation contributed to the overexpression of Dicer1 and the inhibition of tumor growth.

CONCLUSIONS

EBV-miR-BART6-5p may target to facilitate proliferation and metastasis of NPC cells and suppress apoptosis, thus being a new target for NPC therapy.

摘要

目的

探讨 Epstein-Barr 病毒(EBV)编码的 BamHI A 右向转录物(BART)microRNA(miR-BART6-5p)通过靶向 对鼻咽癌(NPC)生物学特性和放射敏感性的影响。

方法

收集 2010 年 1 月至 2011 年 1 月接受放疗的 NPC 患者(n=96),采用实时定量 PCR(qRT-PCR)和 Western blot 检测 miR-BART6-5p 和 Dicer1 的表达。双荧光素酶报告基因实验验证 miR-BART6-5p 靶向 。CCK8、划痕愈合、Transwell 和 Annexin-FITC/PI 实验评估 miR-BART6-5p 介导的 Dicer1 对 NPC 细胞生物学特性的影响。评估 miR-BART6-5p 靶向 的放射敏感性。

结果

NPC 患者中发现 miR-BART6-5p 增加, 减少,与 T 分期、临床分期和 NPC 前 DNA 密切相关。而有效放疗后 NPC 患者中发现 miR-BART6-5p 及 Dicer1 高表达下调。miR-BART6-5p 和 Dicer1 都是 NPC 的预后因素。下调 miR-BART6-5p 可增强 Dicer1 的表达,抑制 NPC 细胞的增殖、侵袭和迁移,促进细胞凋亡。克隆形成实验也表明 miR-BART6-5p 下调降低种植效率(PE),随着照射剂量的增加,PE 进一步降低。6-Gy 照射后向裸鼠注射 miR-BART6-5p 抑制剂促进 Dicer1 过表达并抑制肿瘤生长。

结论

EBV-miR-BART6-5p 可能通过靶向 促进 NPC 细胞的增殖和转移,抑制细胞凋亡,从而成为 NPC 治疗的新靶点。

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