Poodineh Jafar, Sirati-Sabet Majid, Rajabibazl Masoumeh, Mohammadi-Yeganeh Samira
Department of Clinical Biochemistry, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Heliyon. 2020 Nov 9;6(11):e05434. doi: 10.1016/j.heliyon.2020.e05434. eCollection 2020 Nov.
Aberrant Wnt signaling cascade is a hallmark of the triple-negative breast cancer (TNBC) that is linked with the increased proliferation, invasion, and poor overall survival. many genes are post-transcriptionally regulated by microRNAs (miRNAs) therefore; it is indisputable that the dysregulation of the miRNAs is an explanation for the aberrant signaling cascades. Thus, the present study was conducted to find the putative miRNA targeting the key players of Wnt/β -catenin cascade in the TNBC.
The miR-130a-3p was found as a potential regulator of the Wnt signaling cascade by applying several bioinformatic algorithms. Quantitative real-time PCR (qRT-PCR) was used to analyze the expression levels of miR-130a-3p and Wnt cascade genes in the TNBC cells. Afterward, TNBC cells were transiently transfected with the miR-130a-3p to investigate its effects on the expression of Wnt cascade genes. Subsequently, MTT, soft agar colony formation, scratch, transwell cell migration, and transwell cell invasion assays were used to determine the behavior of the TNBC cells in response to miR-130a-3p restoration.
Results of the qRT-PCR showed downregulation of miR-130a-3p and upregulation of the Wnt cascade genes in the TNBC cells compared to the normal cells. Transient overexpression of miR-130a-3p decreased the expression levels of Wnt cascade genes significantly in the TNBC cells. Moreover, following the miR-130a-3p overexpression, the proliferation, anchorage-independent growth, and migration of the TNBC cells were reduced.
Overall, our findings provided an evidence for the significant role of miR-130a-3p in the regulation of Wnt/β-catenin cascade, and also introduced the miR-130a-3p as a new therapeutic target for the patients with TNBC.
异常的Wnt信号级联是三阴性乳腺癌（TNBC）的一个标志，与增殖增加、侵袭以及总体生存率低相关。因此，许多基因受到微小RNA（miRNA）的转录后调控；miRNA失调是异常信号级联的一个解释，这是无可争议的。因此，本研究旨在寻找靶向TNBC中Wnt/β -连环蛋白级联关键分子的假定miRNA。
通过应用多种生物信息学算法，发现miR-130a-3p是Wnt信号级联的潜在调节因子。定量实时聚合酶链反应（qRT-PCR）用于分析TNBC细胞中miR-130a-3p和Wnt级联基因的表达水平。随后，用miR-130a-3p瞬时转染TNBC细胞，以研究其对Wnt级联基因表达的影响。随后，采用MTT、软琼脂集落形成、划痕、Transwell细胞迁移和Transwell细胞侵袭试验来确定TNBC细胞对miR-130a-3p恢复的反应行为。
qRT-PCR结果显示，与正常细胞相比，TNBC细胞中miR-130a-3p下调，Wnt级联基因上调。miR-130a-3p的瞬时过表达显著降低了TNBC细胞中Wnt级联基因的表达水平。此外，miR-130a-3p过表达后，TNBC细胞的增殖、非锚定依赖性生长和迁移减少。
总体而言，我们的研究结果为miR-130a-3p在Wnt/β-连环蛋白级联调节中的重要作用提供了证据，并且还将miR-130a-3p作为TNBC患者的一个新的治疗靶点。
