Browsing by Author "Shuaib, Mohd."

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    Withaferin A mediated changes of miRNA expression in breast cancer-derived mammospheres
    (John Wiley and Sons Inc, 2022-06-30T00:00:00) Prajapati, Kumari Sunita; Shuaib, Mohd.; Gupta, Sanjay; Kumar, Shashank
    Breast cancer is a heterogeneous disease consisting of atypical cell populations that share stem cell-like characteristics associated with therapeutic resistance, disease relapse, and poor clinical outcome. MicroRNAs (miRNA),�and small noncoding RNA, are pivotal in the regulation of self-renewal, stemness, and cellular differentiation. Withaferin A (WA), a steroidal lactone, is a major bioactive constituent of Withania somnifera (Solanaceae) known for its anticancer properties. In this study, the effect of WA on modulation of miRNA expression in breast cancer-derived mammosphere was assessed utilizing small RNA sequencing. Treatment with WA inhibited MCF-7 and T47D cells derived mammosphere formation with a significant decrease in CD44, EpCAM, Nanog, OCT4, and SOX2 as markers of self-renewal and stemness. Small RNA sequencing demonstrated a total of 395 differentially expressed miRNAs (DEMs) including 194 upregulated and 201 downregulated miRNAs in WA-treated�MCF-7 mammospheres. Bioinformatics analysis utilizing the�KEGG pathway, Gene Ontology enrichment, protein?protein, and miRNA-mRNA interaction network identified altered expression in a few hub genes viz.�AKT1, PTEN, MYC, CCND1,�VEGFA,�NOTCH1, and�IGFR1�associated with DEMs in WA-treated�mammospheres. Further quantitative�RT-PCR analysis validated the expression of DEMs including miR-549a-5p, miR-1247-5p, miR-124-5p, miR-137-5p, miR-34a-5p, miR-146a-5p, miR-99a-5p, miR-181a-5p, let-7c-5p, and let-7a-5p. In particular, let-7c-5p is designated as a tumor suppressor in breast cancer. An increase in miR-let-7c-5p expression was noted after WA treatment, with a simultaneous decrease in CCND1 and c-MYC at mRNA and protein levels. Taken together, our study demonstrated WA-mediated miRNA expression, in particular, upregulation of miR-let-7c-5p, leads to the inhibition of breast cancer cells derived mammospheres. � 2022 Wiley Periodicals LLC.