Department of Pediatric Surgery, the First Hospital of Jilin University, Changchun, China.
Eur Rev Med Pharmacol Sci. 2019 Feb;23(3):1030-1037. doi: 10.26355/eurrev_201902_16990.
The aim of this study was to explore the role of microRNA-155-5p (miR-155-5p) in regulating the proliferation and apoptosis of Wilm's tumor (WT), and to investigate the possible underlying mechanism.
The expression levels of miR-155-5p in 37 pairs of WT clinical samples, as well as WT cell line (G401), were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay and flow cytometry assay were used to detect the effects of miR-155-5p on cell proliferation, cycle and apoptosis. Target gene prediction software was applied to screen the potential downstream target gene of miR-155-5p. QRT-PCR, Western blot (WB) and luciferase reporter gene assay proved that cAMP-response element binding protein 1 (CREB1) was the target gene of miR-155-5p. Besides, rescue experiment was conducted to further explore the effect of CREB1 on WT cells.
The expression levels of miR-155-5p in WT tissues and cells were both significantly down-regulated. Importantly, miR-155-5p was found to be involved in the malignant behavior of WT cells. MTT assay and flow cytometry assay demonstrated that miR-155-5p significantly inhibited the proliferation, caused stagnation of cells in G0/G1 phase, and promoted cell apoptosis. CREB1 was verified as a functional target gene of miR-155-5p, which was negatively regulated by miR-155-5p. Rescue experiments indicated that restoring the expression of CREB1 could interfere with the effects of miR-155-5p on WT cells.
MiR-155-5p could regulate the proliferation, cell cycle and apoptosis of WT cells. These effects were achieved by regulating the expression of CREB1. Furthermore, our study might provide a new theoretical basis for the basic research of WT.
本研究旨在探讨微小 RNA-155-5p（miR-155-5p）在调节肾母细胞瘤（WT）增殖和凋亡中的作用，并探讨其可能的潜在机制。
通过定量逆转录聚合酶链反应（qRT-PCR）检测 37 对 WT 临床样本和 WT 细胞系（G401）中 miR-155-5p 的表达水平。MTT（3-(4,5-二甲基噻唑-2-基）-2,5-二苯基四唑溴盐）检测和流式细胞术检测 miR-155-5p 对细胞增殖、周期和凋亡的影响。靶基因预测软件用于筛选 miR-155-5p 的潜在下游靶基因。qRT-PCR、Western blot（WB）和荧光素酶报告基因检测证实 cAMP 反应元件结合蛋白 1（CREB1）是 miR-155-5p 的靶基因。此外，进行了挽救实验以进一步探讨 CREB1 对 WT 细胞的影响。
miR-155-5p 在 WT 组织和细胞中的表达均显著下调。重要的是，miR-155-5p 参与了 WT 细胞的恶性行为。MTT 检测和流式细胞术检测表明，miR-155-5p 可显著抑制细胞增殖，导致细胞停滞在 G0/G1 期，并促进细胞凋亡。CREB1 被验证为 miR-155-5p 的功能靶基因，其受 miR-155-5p 负调控。挽救实验表明，恢复 CREB1 的表达可干扰 miR-155-5p 对 WT 细胞的作用。
miR-155-5p 可调节 WT 细胞的增殖、细胞周期和凋亡。这些作用是通过调节 CREB1 的表达来实现的。此外，我们的研究可能为 WT 的基础研究提供新的理论依据。
