The stemness property of cells allows them to sustain their lineage, differentiation, proliferation, and regeneration. MicroRNAs are small non-coding RNAs known to regulate the stemness property of cells by regulating the expression of stem cell signaling pathway proteins at mRNA level. Dysregulated miRNA expression and associated stem cell signaling pathways in normal stem cells give rise to cancer stem cells. Thus, the present study was aimed to identify the miRNAs involved in the regulation of major stem cell signaling pathways. The proteins (n = 36) involved in the signaling pathways viz., Notch, Wnt, JAK-STAT, and Hedgehog which is associated with the stemness property was taken into the consideration. The miRNAs, having binding sites for the targeted protein-encoding gene were predicted using an online tool (TargetScan) and the common miRNA among the test pathways were identified using Venn diagram analysis. A total of 22 common miRNAs (including 8 non-studied miRNAs) were identified which were subjected to target predictions, KEGG pathway, and gene ontology (GO) analysis to study their potential involvement in the stemness process. Further, we studied the clinical relevance of the non-studied miRNAs by performing the survival analysis and their expression levels in clinical breast cancer patients using the TCGA database. The identified miRNAs showed overall poor survival in breast cancer patients. The miR-6844 showed significantly high expression in various clinical subgroups of invasive breast cancer patients compared with the normal samples. The expression levels of identified miRNA(s) were validated in breast normal, luminal A, triple-negative, and stem cells in vitro models using qRT-PCR analysis. Further treatment with the phytochemical showed excellent down regulation of the lead miRNA. Overall the study first time reports the association of four miRNAs (miR-6791, miR-4419a, miR-4251 and miR-6844) with breast cancer stemness.

细胞的干性特性使它们能够维持其谱系、分化、增殖和再生。MicroRNAs 是小的非编码 RNA，已知通过在 mRNA 水平上调节干细胞信号通路蛋白的表达来调节细胞的干性特性。正常干细胞中失调的 miRNA 表达和相关的干细胞信号通路会产生癌症干细胞。因此，本研究旨在鉴定参与调节主要干细胞信号通路的miRNA。蛋白质（n = 36) 涉及与干性特性相关的信号通路，即 Notch、Wnt、JAK-STAT 和 Hedgehog。使用在线工具 (TargetScan) 预测具有靶向蛋白质编码基因的结合位点的 miRNA，并使用维恩图分析鉴定测试途径中的常见 miRNA。共鉴定出 22 种常见 miRNA（包括 8 种未研究的 miRNA），对其进行靶点预测、KEGG 通路和基因本体（GO）分析，以研究它们在干性过程中的潜在参与。此外，我们通过使用 TCGA 数据库在临床乳腺癌患者中进行生存分析及其表达水平，研究了未研究的 miRNA 的临床相关性。鉴定出的 miRNA 在乳腺癌患者中显示出总体较差的生存率。与正常样本相比，miR-6844 在浸润性乳腺癌患者的各种临床亚组中表现出显着的高表达。使用 qRT-PCR 分析在乳腺正常、管腔 A、三阴性和干细胞体外模型中验证了已识别 miRNA 的表达水平。用植物化学物质进一步处理显示出对先导 miRNA 的极好的下调。总体而言，该研究首次报告了四种 miRNA（miR-6791、miR-4419a、miR-4251 和 miR-6844）与乳腺癌干性的关联。使用 qRT-PCR 分析在乳腺正常、管腔 A、三阴性和干细胞体外模型中验证了已识别 miRNA 的表达水平。用植物化学物质进一步处理显示出对先导 miRNA 的极好的下调。总体而言，该研究首次报告了四种 miRNA（miR-6791、miR-4419a、miR-4251 和 miR-6844）与乳腺癌干性的关联。使用 qRT-PCR 分析在乳腺正常、管腔 A、三阴性和干细胞体外模型中验证了已识别 miRNA 的表达水平。用植物化学物质进一步处理显示出对先导 miRNA 的极好的下调。总体而言，该研究首次报告了四种 miRNA（miR-6791、miR-4419a、miR-4251 和 miR-6844）与乳腺癌干性的关联。