Department of Clinical Biochemistry, School of Medicine, Zabol University of Medical Sciences, Zabol, Iran.
Department of Clinical Biochemistry, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Chem Biol Drug Des. 2022 Sep;100(3):334-345. doi: 10.1111/cbdd.14113. Epub 2022 Jul 14.
The Notch-regulated ankyrin repeat protein (NRARP) functions as a molecular link between Notch and Wnt signaling pathways. Although it has recently been identified to be overexpressed in breast cancer (BC), the molecular mechanisms that regulate NRARP remain unknown. Since microRNAs (miRNAs) regulate gene expression post-transcriptionally, miRNA dysregulation could explain the abnormal gene expression. Here, we identified miR-130a-3p as an NRARP regulator and evaluated its effects on the behavior of BC cells. Quantitative real-time PCR was performed to assess the transcriptional levels of miR-130a-3p and NRARP in BC cells. Next, miR-130a-3p was transiently transfected into BC cells to assess its influence on NRARP expression. Owing to the positive regulatory effects of NRARP on the Wnt/β-catenin signaling pathway, we also analyzed the expression levels of five Wnt/β-catenin pathway genes and one downstream target gene in BC cells. We then assessed anti-tumor activities of miR-130a-3p in BC cells using the MTT proliferation assay, the soft agar colony formation assay for anchorage-independent growth (AIG), as well as scratch and transwell assays for cell migration. The results showed that miR-130a-3p was downregulated in BC cells, whereas NRARP was upregulated. Overexpression of miR-130a-3p inhibited the expression of NRARP and some Wnt/β-catenin signaling pathway genes, as well as exerted anti-tumor effects as evidenced by decreased cell proliferation, AIG, and migration of BC cells. In conclusion, the tumor-suppressive function of miR-130a-3p in BC may be mediated by inhibiting NRARP and Wnt/β-catenin signaling pathway. As a result, miR-130a-3p could be introduced as a therapeutic target for miRNA therapy in BC.
Notch 调控锚蛋白重复蛋白（NRARP）作为 Notch 和 Wnt 信号通路之间的分子连接物发挥作用。尽管最近已经确定它在乳腺癌（BC）中过表达，但调节 NRARP 的分子机制仍然未知。由于 microRNAs（miRNAs）在后转录水平上调节基因表达，miRNA 失调可能解释异常的基因表达。在这里，我们确定 miR-130a-3p 为 NRARP 的调节剂，并评估其对 BC 细胞行为的影响。定量实时 PCR 用于评估 BC 细胞中 miR-130a-3p 和 NRARP 的转录水平。接下来，将 miR-130a-3p 瞬时转染到 BC 细胞中，以评估其对 NRARP 表达的影响。由于 NRARP 对 Wnt/β-catenin 信号通路具有正调节作用，我们还分析了 BC 细胞中五个 Wnt/β-catenin 通路基因和一个下游靶基因的表达水平。然后，我们使用 MTT 增殖测定法、用于锚定独立生长（AIG）的软琼脂集落形成测定法以及划痕和 Transwell 测定法评估 miR-130a-3p 在 BC 细胞中的抗肿瘤活性。结果表明，miR-130a-3p 在 BC 细胞中下调，而 NRARP 上调。miR-130a-3p 的过表达抑制了 NRARP 和一些 Wnt/β-catenin 信号通路基因的表达，并通过降低 BC 细胞的增殖、AIG 和迁移来发挥抗肿瘤作用。总之，miR-130a-3p 在 BC 中的肿瘤抑制功能可能是通过抑制 NRARP 和 Wnt/β-catenin 信号通路介导的。因此，miR-130a-3p 可以作为 BC 中 miRNA 治疗的治疗靶点。
