miRNA-548c-5p 通过调控 Wnt/β-连环蛋白信号通路抑制乳腺癌细胞的增殖。
MicroRNA-548c-5p inhibits the proliferation of breast cancer cells through regulating Wnt/β-catenin signaling pathway.
机构信息
Department of Oncology, Baoan Central Hospital of Shenzhen, the 5th Affiliated Hospital of Shenzhen University Health Science Center, Shenzhen, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Apr;24(7):3795-3804. doi: 10.26355/eurrev_202004_20845.
OBJECTIVE
The aim of this study was to explore the function of microRNA-548c-5p in breast cancer (BCa) and the underlying mechanism. Our findings might help to provide a theoretical basis for the diagnosis and treatment of BCa.
PATIENTS AND METHODS
Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression level of microRNA-548c-5p in BCa tumor tissues and para-cancerous tissues. The relationship between microRNA-548c-5p expression and clinical indicators of BCa was analyzed. Meanwhile, the expression of microRNA-548c-5p in the BCa cells was detected by qRT-PCR as well. MicroRNA-548c-5p overexpression and the knockdown models were constructed in BCa cell lines MCF-7 and MDA-MB-231. Subsequently, Cell Counting Kit-8 (CCK-8), colony formation and 5-ethynyl-2'-deoxyuridine (EdU) assays were performed to analyze the influence of microRNA-548c-5p on biological functions of BCa cells. Finally, the interaction between microRNA-548c-5p and Wnt/b-catenin signaling pathway was investigated.
RESULTS
QRT-PCR results showed that the expression level of microRNA-548c-5p in BCa tumor tissues was remarkably lower than that of adjacent tissues, and the difference was statistically significant (p<0.05). Compared with patients with high expression of microRNA-548c-5p, the pathological stage of patients with low microRNA-548c-5p expression was significantly higher (p<0.05). Similarly, microRNA-548c-5p overexpression remarkably decreased the proliferation ability of BCa cells in vitro. However, microRNA-548c-5p knockdown showed an opposite trend. In addition, Wnt1, a key factor in the Wnt/b-catenin signaling pathway, was found remarkably up-regulated in BCa cell lines and tissues. Wnt1 expression was negatively correlated with microRNA-548c-5p expression. Western Blotting results demonstrated that microRNA-548c-5p mimics remarkably down-regulated the levels of the proteins in the Wnt/b-catenin signaling pathway. Conversely, opposite results were observed in microRNA-548c-5p inhibitor group. The rescue experiments in the cells revealed that there might be a mutual regulation between microRNA-548c-5p and Wnt1, thereby together regulating the malignant growth of BCa.
CONCLUSIONS
MicroRNA-548c-5p was lowly expressed in BCa tissues and cells, which was closely related to the pathological stage of BCa. In addition, microRNA-548c-5p significantly inhibited the proliferation of BCa cells via modulating Wnt/b-catenin signaling pathway.
目的
本研究旨在探讨微小 RNA-548c-5p 在乳腺癌（BCa）中的功能及其潜在机制。我们的研究结果可能有助于为 BCa 的诊断和治疗提供理论依据。
方法
采用实时定量聚合酶链反应（qRT-PCR）检测 BCa 肿瘤组织和癌旁组织中微小 RNA-548c-5p 的表达水平。分析微小 RNA-548c-5p 表达与 BCa 临床指标的关系。同时，采用 qRT-PCR 检测 BCa 细胞中微小 RNA-548c-5p 的表达。在 MCF-7 和 MDA-MB-231 细胞系中构建微小 RNA-548c-5p 过表达和敲低模型。随后，通过细胞计数试剂盒-8（CCK-8）、集落形成和 5-乙炔基-2'-脱氧尿苷（EdU）检测分析微小 RNA-548c-5p 对 BCa 细胞生物学功能的影响。最后，研究微小 RNA-548c-5p 与 Wnt/β-catenin 信号通路的相互作用。
结果
qRT-PCR 结果显示，BCa 肿瘤组织中微小 RNA-548c-5p 的表达水平明显低于邻近组织，差异具有统计学意义（p<0.05）。与微小 RNA-548c-5p 高表达患者相比，微小 RNA-548c-5p 低表达患者的病理分期明显更高（p<0.05）。同样，微小 RNA-548c-5p 过表达显著降低了 BCa 细胞在体外的增殖能力。然而，微小 RNA-548c-5p 敲低则呈现出相反的趋势。此外，Wnt1，Wnt/β-catenin 信号通路中的关键因子，在 BCa 细胞系和组织中表达明显上调。Wnt1 表达与微小 RNA-548c-5p 表达呈负相关。Western Blotting 结果表明，微小 RNA-548c-5p 模拟物显著下调了 Wnt/β-catenin 信号通路中蛋白的水平。相反，微小 RNA-548c-5p 抑制剂组则观察到相反的结果。细胞中的挽救实验表明，微小 RNA-548c-5p 和 Wnt1 之间可能存在相互调节作用，从而共同调节 BCa 的恶性生长。
结论
微小 RNA-548c-5p 在 BCa 组织和细胞中低表达，与 BCa 的病理分期密切相关。此外，微小 RNA-548c-5p 通过调节 Wnt/β-catenin 信号通路显著抑制 BCa 细胞的增殖。
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