Department of Surgery, The Second Affiliated Hospital of Soochow University, China.
Department of General Surgery, The First People's Hospital of Nantong, China.
J Gene Med. 2020 Aug;22(8):e3188. doi: 10.1002/jgm.3188. Epub 2020 Apr 13.
Breast cancer is the leading cause of cancer deaths in women worldwide. The purpose of the current study was to investigate the potential role of miR-96-5p in breast cancer.
Breast cancer tissues and matched para-cancerous tissues were collected. The expression of microRNA-96-5p (miR-96-5p) and arginine kinase 3 (AK3) was detected by quantitative real-time PCR (qRT-PCR). The correlation between miR-96-5p and AK3 was calculated by Pearson's Chi-square test. Moreover, mimics or inhibitors of miR-96-5p were applied to explore whether miR-96-5p influences the migration capacity in Transwell and wound healing assays. Bioinformatics analysis was performed to identify the target genes of miR-96-5p through the TargetScan, miRDB and miRanda databases. A luciferase reporter assay was performed to verify AK3 as a downstream target gene of miR-96-5p.
The expression of miR-96-5p was significantly increased in breast cancer tissue and breast cancer cell lines compared with para-cancerous tissue and a breast cell line, respectively. The expression of miR-96-5p negatively correlated with AK3 gene expression. AK3 was demonstrated to be a direct mRNA target of miR-96-5p. AK3 was positively associated with the overall survival of breast cancer patients. Kaplan-Meier curve and log rank test analyses revealed that decreased AK3 levels were significantly associated with reduced overall survival. miR-96-5p was shown to promote the migration of breast cancer cells through the MEK/ERK signaling pathway.
Our results identify a role for miR-96-5p in promoting breast cancer cell migration through activation of MEK/ERK signaling by targeting AK3.
乳腺癌是全球女性癌症死亡的主要原因。本研究旨在探讨 microRNA-96-5p（miR-96-5p）在乳腺癌中的潜在作用。
收集乳腺癌组织和配对癌旁组织。采用实时定量 PCR（qRT-PCR）检测 microRNA-96-5p（miR-96-5p）和精氨酸激酶 3（AK3）的表达。采用 Pearson's Chi-square 检验计算 miR-96-5p 与 AK3 的相关性。此外，应用 miR-96-5p 的模拟物或抑制剂，通过 Transwell 和划痕愈合实验探讨 miR-96-5p 是否影响迁移能力。通过 TargetScan、miRDB 和 miRanda 数据库进行生物信息学分析，以确定 miR-96-5p 的靶基因。通过荧光素酶报告基因实验验证 AK3 是 miR-96-5p 的下游靶基因。
与癌旁组织和乳腺细胞系相比，乳腺癌组织和乳腺癌细胞系中 miR-96-5p 的表达明显增加。miR-96-5p 的表达与 AK3 基因表达呈负相关。AK3 被证明是 miR-96-5p 的直接 mRNA 靶基因。AK3 与乳腺癌患者的总生存期呈正相关。Kaplan-Meier 曲线和对数秩检验分析表明，AK3 水平降低与总生存期缩短显著相关。miR-96-5p 通过 MEK/ERK 信号通路促进乳腺癌细胞的迁移。
本研究结果表明，miR-96-5p 通过靶向 AK3 激活 MEK/ERK 信号通路，在促进乳腺癌细胞迁移中发挥作用。
