miR-451a 通过 Ras/Raf/MEK/ERK 通路靶向 RAB5A 抑制高级别浆液性卵巢癌的增殖和迁移。

miR-451a suppresses the proliferation and migration of high-grade serous ovarian cancer by targeting RAB5A through the Ras/Raf/MEK/ERK pathway.

机构信息

Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, Jinan, Shandong, China.

Department of Gynecology, Shandong Provincial Hospital affiliated to Shandong First Medical University, Jinan, China.

出版信息

J Gene Med. 2024 Jan;26(1):e3649. doi: 10.1002/jgm.3649.

DOI:10.1002/jgm.3649

PMID:38282155

Abstract

BACKGROUND

Ovarian cancer is one of the most common cancers in women. Profiles changes of microRNAs (miRNAs) are closely linked to malignant tumors. In the present study, we investigated expression of miR-451a in high-grade serous ovarian cancer (HGSOC). We also investigated the potential pathological roles and the likely mechanism of miR-451a in the development of HGSOC using animal models and cell lines.

METHODS

Using bioinformatics techniques and a real-time PCR, we analyzed differently expressed miRNAs in HGSOC compared to normal tissue. MTT (i.e. 3-[4, 5-dimethyl thiazol-2-yl]-2,5-diphenyl tetrazolium bromide), EDU (i.e. 5-ethynyl-2'-deoxyuridine) and transwell assays were performed to investigate the effect of miR-451a on the proliferation and migration of HGSOC SKOV-3 cells. A dual luciferase reporter assay was performed to verify the targeting relationship of miR-451 and RAB5A (one of the Rab GTPase proteins that regulates endocytosis and vesicle transport). Also, we analyzed levels of the RAB5A mRNA and protein by real-time PCR, western blotting and immunohistochemistry assays in HGSOC cells and tissues. Finally, we performed in vivo experiments using HGSOC mice.

RESULTS

miR-451a was substantially upregulated in HGSOC and associated with favorable clinical characteristics. miR-451a knockdown significantly increased growth and metastasis of HGSOC cell line SKOV-3 through Ras/Raf/mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) signaling. In addition, RAB5A, an early endosome marker, was shown to be a direct target of miR-451a. Moreover, RAB5A is correlated with unfavorable clinical features and shows independent prognostic significance in HGSOC.

CONCLUSIONS

We found that the miR-451a/RAB5A axis is associated with tumorigenesis and progression through the Ras/Raf/MEK/ERK pathway, providing prognostic indicators and therapeutic targets for patients with HGSOC.

摘要

背景

卵巢癌是女性最常见的癌症之一。miRNA（miRNA）的表达谱变化与恶性肿瘤密切相关。在本研究中，我们研究了 miR-451a 在高级别浆液性卵巢癌（HGSOC）中的表达。我们还使用动物模型和细胞系研究了 miR-451a 在 HGSOC 发展中的潜在病理作用及其可能的机制。

方法

使用生物信息学技术和实时 PCR 分析 HGSOC 与正常组织中差异表达的 miRNA。MTT（即 3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四唑溴盐）、EDU（即 5-乙炔基-2'-脱氧尿苷）和 Transwell 测定法用于研究 miR-451a 对 HGSOC SKOV-3 细胞增殖和迁移的影响。双荧光素酶报告基因测定法用于验证 miR-451 和 RAB5A（一种调节内吞作用和囊泡运输的 Rab GTPase 蛋白之一）之间的靶向关系。还通过实时 PCR、western blot 和免疫组织化学测定法分析了 HGSOC 细胞和组织中 RAB5A mRNA 和蛋白的水平。最后，我们使用 HGSOC 小鼠进行了体内实验。

结果

miR-451a 在 HGSOC 中大量上调，与良好的临床特征相关。miR-451a 敲低通过 Ras/Raf/丝裂原活化蛋白激酶激酶（MEK）/细胞外信号调节激酶（ERK）信号通路显著增加 HGSOC 细胞系 SKOV-3 的生长和转移。此外，早期内体标志物 RAB5A 被证明是 miR-451a 的直接靶标。此外，RAB5A 与不良的临床特征相关，并在 HGSOC 中具有独立的预后意义。