环状 RNA-ABCB10 通过调控 miR-620/FABP5 轴在神经胶质瘤细胞中发挥癌基因作用。
Circ-ABCB10 acts as an oncogene in glioma cells via regulation of the miR-620/FABP5 axis.
机构信息
Department of Neurology, The Fifth Affiliated Hospital of Wenzhou Medical University, Lishui, Zhejiang, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Jun;24(12):6848-6857. doi: 10.26355/eurrev_202006_21674.
OBJECTIVE
This study aims to investigate the biological function of circular RNA ABCB10 (circ-ABCB10) in regulating the progression of glioma and to study the possible underlying mechanisms.
PATIENTS AND METHODS
The expression levels of circ-ABCB10, miR-620 and FABP5 mRNA in glioma tissues, normal surrounding tissues and glioma cell lines were measured by Real-time PCR (RT-PCR). Circ-ABCB10 was silenced by siRNA in glioma cell lines (U87, T98G). The proliferation, migration and invasion of glioma cells were measured by MTT, wound healing and transwell assays, respectively. The relationship between circ-ABCB10, miR-620 and FABP5 was tested by Dual-Luciferase assay. The expression of proteins was measured by Western blot. The cell cycle distribution and apoptosis were measured by flow cytometry.
RESULTS
The expression levels of circ-ABCB10 and FABP5 in glioma tissues and cells were significantly higher than those in their normal counterparts. Moreover, the expression of miR-620 was lower in glioma tissues. Silencing of circ-ABCB10 in glioma cells significantly inhibited the proliferation, migration and invasion of glioma cells. Moreover, downregulation of circ-ABCB10 induced cell cycle arrest and apoptosis in glioma cells. Furthermore, inhibition of miR-620 showed the opposite effects to silencing circ-ABCB10 on glioma cells. Dual-Luciferase reporter assays demonstrated that circ-ABCB10 could bind to miR-620 and that FABP5 was a direct target of miR-620. Western blot results showed that circ-ABCB10 could stabilize the expression of FABP5, while miR-620 decreased the expression of FABP5. Furthermore, overexpression of FABP5 abrogated the silencing effects of circ-ABCB10 in glioma cells.
CONCLUSIONS
These data suggest that circ-ABCB10 affects glioma progression by regulating the miR-620/FABP5 axis, and circ-ABCB10 might be used as a potential target for the treatment of glioma.
目的
本研究旨在探讨 ABCB10 环状 RNA（circ-ABCB10）在调控神经胶质瘤进展中的生物学功能，并研究其可能的作用机制。
方法
采用实时荧光定量 PCR（RT-PCR）检测神经胶质瘤组织、正常周围组织和神经胶质瘤细胞系中 circ-ABCB10、miR-620 和 FABP5mRNA 的表达水平。用 siRNA 沉默神经胶质瘤细胞系（U87、T98G）中的 circ-ABCB10。采用 MTT、划痕愈合和 Transwell 实验分别检测神经胶质瘤细胞的增殖、迁移和侵袭能力。通过双荧光素酶报告基因实验检测 circ-ABCB10、miR-620 与 FABP5 之间的关系。采用 Western blot 检测蛋白表达。采用流式细胞术检测细胞周期分布和凋亡。
结果
神经胶质瘤组织和细胞中 circ-ABCB10 和 FABP5 的表达水平明显高于其正常对应物。此外，神经胶质瘤组织中 miR-620 的表达水平较低。沉默神经胶质瘤细胞中的 circ-ABCB10 可显著抑制神经胶质瘤细胞的增殖、迁移和侵袭。此外，下调 circ-ABCB10 可诱导神经胶质瘤细胞发生细胞周期阻滞和凋亡。此外，抑制 miR-620 对神经胶质瘤细胞的作用与沉默 circ-ABCB10 的作用相反。双荧光素酶报告基因实验表明，circ-ABCB10 可与 miR-620 结合，而 FABP5 是 miR-620 的直接靶标。Western blot 结果表明，circ-ABCB10 可稳定 FABP5 的表达，而 miR-620 则降低 FABP5 的表达。此外，过表达 FABP5 可消除 circ-ABCB10 在神经胶质瘤细胞中的沉默作用。
结论
这些数据表明，circ-ABCB10 通过调节 miR-620/FABP5 轴影响神经胶质瘤的进展，circ-ABCB10 可能作为神经胶质瘤治疗的潜在靶点。
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