Comprehensive phytochemical profiling and cytotoxic assessment of Caucasalia macrophylla coupled with in silico docking and miRNA (miR-19b, miR-20a, miR-126, miR-155, miR-200c)-Mediated gene regulation analysis.

Abstract

This study aimed to identify the chemical constituents of the methanol extract of Caucasalia macrophylla using GC-MS and HPLC-DAD techniques. The cytotoxic activity was determined by the MTT assay using human breast adenocarcinoma cells (MDA-MB-231) and hTERT-immortalized mammary epithelial cells (hTERT-HME1). The half-maximal inhibitory concentration (IC₅₀) for MDA-MB-231 cells was found to be 1.2 mg/ml. This concentration inhibited cell proliferation in a concentration-dependent manner. In addition, the extract was tested using a miRNA Isolation Kit to assess the mRNA expression profiles in both MDA-MB-231 and hTERT-HME1 cell lines. In cancer cells (MDA-MB-231), the expression of miR-126 was significantly upregulated 2.2-fold, while miR-155 and miR-200c were significantly downregulated 0.3-fold and 0.4-fold respectively. In healthy cells (hTERT-HME1), the expression levels of miR-19b, miR-155, and miR-200c were also significantly reduced. Further a comprehensive molecular docking analysis was conducted against phosphoinositide 3-kinase (PI3K). Among the tested compounds, rutin exhibited the strongest binding affinity with a binding energy of - 9.1 kcal/mol. This compound demonstrated potential anticancer effects by modulating PI3K activity. The findings suggest that modulation of signaling pathways by natural compounds may represent a promising approach for development of novel cancer therapies.