Synergistic Effect of Curcuma longa Extract in Combination with Phyllanthus niruri Extract in Regulating Annexin A2, Epidermal Growth Factor Receptor, Matrix Metalloproteinases, and Pyruvate Kinase M1/2 Signaling Pathway on Breast Cancer Stem Cell

Abstract

AIM: This study aimed to investigate the synergistic effects of the combination between Curcuma longa extract (CL) and Phyllanthus niruri extract (PN) in inhibiting optimally the MDA-MB-231 breast cancer stem cells (BCSCs) growth and metastatic by exploring the target and molecular mechanism using integrative bioinformatics approaches and in vitro. METHODS: CL and PN extracts were prepared by maceration method using ethanol 70%. The antiproliferative effect of CL and PN single and combination treatment was examined by 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bromide assay. The bioinformatic approach was performed to identify molecular targets, key proteins, and molecular mechanism of curcumin and phyllanthin as CL and PN secondary metabolite, respectively, targeted at stemness and migration pathway of BCSCs. RESULTS: The in vitro study showed that CL and PN possess cytotoxic activity in time- and dose-dependent manner. The combination of CL and PN has a synergistic effect by modulating the sensitivity of cells. Using a bioinformatics approach, the annexin A2 (ANXA2), epidermal growth factor receptor (EGFR), matrix metalloproteinases (MMPs), and pyruvate kinase M1/2 (PKM) as potential targets of curcumin and phyllanthin correlated with metastatic inhibition of BC. In addition, molecular docking showed that curcumin and phyllanthin performed similar or better interaction to stemness differentiation regulator pathway particularly histone deacetylase 1, EGFR, Heat Shock Protein 90 Alpha Family Class B Member 1, Hypoxia Inducible Factor 1 Subunit Alpha, and MMP9. CONCLUSION: Combination of CL and PN has potential for the treatment of metastatic BCSCs by targeting ANXA2, EGFR, MMPs, and PKM to resolve stemness and inhibit of BCSCs.