环状 RNA ZNF609 通过与 microRNA-150-5p 竞争促进鼻咽癌的生长和转移。

CircRNA ZNF609 promotes growth and metastasis of nasopharyngeal carcinoma by competing with microRNA-150-5p.

机构信息

Department of Otolaryngology Head and Neck Surgery, the Affiliated Hospital of Southwest Medical University, Luzhou, China.

出版信息

Eur Rev Med Pharmacol Sci. 2019 Apr;23(7):2817-2826. doi: 10.26355/eurrev_201904_17558.

DOI:10.26355/eurrev_201904_17558

PMID:31002133

Abstract

OBJECTIVE

This study aims to explore the biological function of circular RNA ZNF609 (circ-ZNF609) in regulating the occurrence and progression of nasopharyngeal carcinoma (NPC), and to investigate the possible underlying mechanism.

PATIENTS AND METHODS

The expression levels of circ-ZNF609, microRNA-150-5p and Sp1 in NPC tissues and normal nasopharyngeal epithelial tissues were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Circ-ZNF609 expression was silenced in NPC cell lines (5-8F and HONE-1). Cellular behaviors of NPC cells were determined using Cell Counting Kit-8 (CCK-8), wound healing and transwell assay. The binding relationship among microRNA-150-5p, circ-ZNF609 and Sp1 was detected by Dual-Luciferase reporter gene assay. In addition, the protein expression of Sp1 after altering expression of circ-ZNF609 or microRNA-150-5p was detected by Western blot.

RESULTS

The expression levels of circ-ZNF609 and Sp1 in NPC tissues were markedly higher than those of normal nasopharyngeal epithelial tissues. However, the expression of microRNA-150-5p was significantly lower in NPC tissues. The knockdown of circ-ZNF609 in NPC cells 5-8F and HONE-1 significantly inhibited the proliferative, migratory and invasive behaviors of NPC cells. Meanwhile, microRNA-150-5p knockdown in NPC cells showed the opposite effect on cellular behaviors of NPC cells. Dual-Luciferase reporter gene assay revealed that circ-ZNF609 could bind to microRNA-150-5p, and Sp1 was a target gene of microRNA-150-5p. Western blot results showed that circ-ZNF609 could stabilize the expression of Sp1, while microRNA-150-5p degraded Sp1 expression. Furthermore, the knockdown of Sp1 in NPC cells reversed the carcinogenic effect of circ-ZNF609.

CONCLUSIONS

Highly expressed circ-ZNF609 adsorbs microRNA-150-5p to upregulate Sp1 expression, thereby promoting the proliferation and metastatic ability of NPC cells.

摘要

目的

本研究旨在探讨环状 RNA ZNF609（circ-ZNF609）在调控鼻咽癌（NPC）发生和发展中的生物学功能，并探讨其可能的潜在机制。

方法

采用实时定量聚合酶链反应（qRT-PCR）检测 NPC 组织和正常鼻咽上皮组织中 circ-ZNF609、miR-150-5p 和 Sp1 的表达水平。在 NPC 细胞系（5-8F 和 HONE-1）中沉默 circ-ZNF609 的表达。采用细胞计数试剂盒-8（CCK-8）、划痕愈合和 Transwell 实验测定 NPC 细胞的细胞行为。通过双荧光素酶报告基因实验检测 miR-150-5p、circ-ZNF609 和 Sp1 之间的结合关系。此外，通过 Western blot 检测改变 circ-ZNF609 或 miR-150-5p 表达后 Sp1 的蛋白表达。

结果

NPC 组织中 circ-ZNF609 和 Sp1 的表达水平明显高于正常鼻咽上皮组织，而 miR-150-5p 在 NPC 组织中的表达明显降低。NPC 细胞 5-8F 和 HONE-1 中 circ-ZNF609 的敲低显著抑制 NPC 细胞的增殖、迁移和侵袭能力。同时，NPC 细胞中 miR-150-5p 的敲低对 NPC 细胞的细胞行为表现出相反的作用。双荧光素酶报告基因实验表明，circ-ZNF609 可与 miR-150-5p 结合，Sp1 是 miR-150-5p 的靶基因。Western blot 结果显示，circ-ZNF609 可稳定 Sp1 的表达，而 miR-150-5p 降解 Sp1 的表达。此外，NPC 细胞中 Sp1 的敲低逆转了 circ-ZNF609 的致癌作用。