miRNA-181a-5p 通过靶向 NRAS 原癌基因抑制视网膜母细胞瘤细胞的增殖、迁移和侵袭。

microRNA-181a-5p impedes the proliferation, migration, and invasion of retinoblastoma cells by targeting the NRAS proto-oncogene.

机构信息

Shenzhen Eye Hospital, Jinan University, China.

出版信息

Clinics (Sao Paulo). 2022 Mar 24;77:100026. doi: 10.1016/j.clinsp.2022.100026. eCollection 2022.

OBJECTIVES

Accumulating research have reported that microRNAs (miRNAs) play important roles in Retinoblastoma (RB). Nonetheless, the function and underlying mechanism of miR-181a-5p in RB remain ambiguous.

METHODS

The relative expression levels of miR-181a-5p and NRAS mRNA were detected by quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). RB cell proliferation was measured using the Cell Counting Kit-8 (CCK-8) and 5'-Bromo-2'-deoxyuridine (BrdU) assays. Transwell assays and flow cytometry were performed to detect the migration, invasion, and apoptosis of RB cells. The interaction between miR-181a-5p and NRAS was explored using luciferase experiments, western blotting, and qRT-PCR.

RESULTS

miR-181a-5p expression was found to be decreased in RB tissues and cell lines, and its expression was correlated with unfavorable pathological features of the patients. In vitro experiments revealed that miR-181a-5p reduced RB cell proliferation, migration, and invasion while enhancing apoptosis. Further research confirmed that NRAS is a direct target of miR-181a-5p. miR-181a-5p inhibited NRAS expression at both the mRNA and protein levels. Co-transfection of pcDNA-NRAS or NRAS small interfering RNA (siRNA) reversed the effects of miR-181a-5p mimics or miR-181a-5p inhibitors on RB cells.

CONCLUSION

miR-181a-5p was significantly downregulated during the development of RB, and it suppressed the malignant behaviors of RB cells by targeting NRAS.

目的

越来越多的研究表明 microRNAs（miRNAs）在视网膜母细胞瘤（RB）中发挥重要作用。然而，miR-181a-5p 在 RB 中的功能和潜在机制仍不清楚。

方法

通过定量逆转录聚合酶链反应（qRT-PCR）检测 miR-181a-5p 和 NRAS mRNA 的相对表达水平。使用细胞计数试剂盒-8（CCK-8）和 5'-溴-2'-脱氧尿苷（BrdU）测定 RB 细胞增殖。通过 Transwell 测定和流式细胞术检测 RB 细胞的迁移、侵袭和凋亡。使用荧光素酶实验、western blot 和 qRT-PCR 研究 miR-181a-5p 和 NRAS 之间的相互作用。

结果

发现 miR-181a-5p 在 RB 组织和细胞系中表达降低，其表达与患者不良的病理特征相关。体外实验表明，miR-181a-5p 降低 RB 细胞增殖、迁移和侵袭，同时增强细胞凋亡。进一步的研究证实 NRAS 是 miR-181a-5p 的直接靶标。miR-181a-5p 在 mRNA 和蛋白水平上均抑制 NRAS 表达。pcDNA-NRAS 或 NRAS 小干扰 RNA（siRNA）的共转染逆转了 miR-181a-5p 模拟物或 miR-181a-5p 抑制剂对 RB 细胞的影响。