Department of Pathology, Basic Medical College, Weifang Medical University, Weifang, China.
Department of Human Anatomy, Basic Medical College, Weifang Medical University, Weifang, China.
Thorac Cancer. 2020 Nov;11(11):3126-3138. doi: 10.1111/1759-7714.13620. Epub 2020 Sep 22.
microRNAs (miRNAs) have been verified as molecular targets for regulating tumor proliferation, invasion, and metastasis in tumor progression. However, the relationship between miRNAs and cellular energy metabolism in breast cancer still needs to be clarified. This study aimed to investigate the role of miR-429 in breast cancer progression.
Bioinformatic analyses were employed to detect the relationship between miR-429 and cancer-related signaling pathways. We used a Kaplan-Meier curve to analyze survival rate in patients with high or low expression of miR-429. We used real-time quantitative PCR (RT-qPCR) to detect the expression of miR-429 in different cell lines. Sh-con, over-miR-429, miR-429 inhibitor, and sh-inhibitor control were transfected. Colony formation and EDU assay were used to detect the proliferation of transfected cells. Wound healing and transwell assays were performed to detect the mobility and invasion ability of transfected cells. Western blot assay was used to detect relative protein expression in transfected cells and different tissues. Bioinformatic analyses were conducted to detect the target proteins expression in different breast cancer databases. Dual luciferase reporter assay was used to confirm the binding site between miR-429 and fibronectin 1 (FN1).
The results of our study indicate that MiR-429 and its target genes are associated with cancer-related signaling pathways and that higher miR-429 expression corresponds with a better prognosis. When miR-429 was overexpressed, the proliferation, invasion of MDA-MB-231 were inhibited. MiR-429 was able to suppress the Wnt/β-catenin signaling pathway, and FN1 overexpression could rescue the influence of over-miR-429.
The results of our study suggest that miR-429 suppresses the proliferation and invasion of breast cancer via inhibiting the Wnt/β-catenin signaling pathway.
微小 RNA（miRNAs）已被证实可作为调控肿瘤增殖、侵袭和转移的分子靶点，从而影响肿瘤的进展。然而，miRNAs 与乳腺癌细胞能量代谢的关系仍有待阐明。本研究旨在探讨 miR-429 在乳腺癌进展中的作用。
采用生物信息学分析检测 miR-429 与癌症相关信号通路的关系。我们使用 Kaplan-Meier 曲线分析 miR-429 高表达或低表达患者的生存率。采用实时定量 PCR（RT-qPCR）检测不同细胞系中 miR-429 的表达。转染 sh-con、过表达 miR-429、miR-429 抑制剂和 sh-inhibitor 对照。使用集落形成和 EDU 检测转染细胞的增殖。进行划痕愈合和 Transwell 检测以检测转染细胞的迁移和侵袭能力。Western blot 检测转染细胞和不同组织中的相对蛋白表达。通过生物信息学分析检测不同乳腺癌数据库中的靶蛋白表达。双荧光素酶报告实验验证 miR-429 与纤维连接蛋白 1（FN1）的结合位点。
我们的研究结果表明，miR-429 及其靶基因与癌症相关信号通路有关，miR-429 表达水平较高与预后较好相关。过表达 miR-429 可抑制 MDA-MB-231 的增殖和侵袭。miR-429 能够抑制 Wnt/β-catenin 信号通路，而 FN1 的过表达可挽救过表达 miR-429 的影响。
本研究结果表明，miR-429 通过抑制 Wnt/β-catenin 信号通路抑制乳腺癌的增殖和侵袭。
