anti-proliferative, apoptotic induction, and anti-migration effects of hemi-synthetic 1′s-1′-acetoxychavicol acetate analogs on mda-mb-231 breast cancer cells

Su Ki Liew,1 Mohamad Nurul Azmi,2 Lionel LA In,3 Khalijah Awang,4,5 Noor Hasima Nagoor1,6 1Institute of Biological Science (Genetics & Molecular Biology), Faculty of Science, University of Malaya, Kuala Lumpur, 2School of Chemical Sciences, Universiti Sains Malaysia, Penang, 3Department of Biotechnology, Faculty of Applied Sciences, UCSI University, 4Centre for Natural Product Research and Drug Discovery (CENAR), 5Department of Chemistry, Faculty of Science, 6Centre for Research in Biotechnology for Agriculture (CEBAR), University of Malaya, Kuala Lumpur, Malaysia Abstract: Nine analogs of 1'S-1'-acetoxychavicol acetate (ACA) were hemi-synthesized and evaluated for their anticancer activities against seven human cancer cell lines. The aim of this study was to investigate the anti-proliferative, apoptotic, and anti-migration effects of these compounds and to explore the plausible underlying mechanisms of action. We found that ACA and all nine analogs were non toxic to human mammary epithelial cells (HMECs) used as normal control cells, and only ACA, 1'-acetoxyeugenol acetate (AEA), and 1'-acetoxy-3,5-dimethoxychavicol acetate (AMCA) inhibited the growth of MDA-MB-231 breast cancer cells with a half-maximal inhibitory concentration (IC50) value of <30.0 µM based on 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay results, and were selected for further investigation. DNA fragmentation assays showed that these three compounds markedly induced apoptosis of MDA-MB-231 cells. Western blot analysis revealed increased expression levels of cleaved PARP, p53, and Bax, while decreased expression levels of Bcl-2 and Bcl-xL were seen after treatment, indicating that apoptosis was induced via the mitochondrial pathway. Moreover, ACA, AEA, and AMCA effectively inhibited the migration of MDA-MB-231 cells. They also downregulated the expression levels of pFAK/FAK and pAkt/Akt via the integrin ß1-mediated signaling pathway. Collectively, ACA and its hemi-synthetic analogs, AEA and AMCA are seen as potential anticancer agents following their abilities to suppress growth, induce apoptosis, and inhibit migration of breast cancer cells. Keywords: 1'S-1'-acetoxychavicol acetate, ACA, ACA hemi-synthetic analogs, triple-negative breast cancer, MTT assay, DNA fragmentation, wound-healing assay, Western blot, integrin β1 signaling pathway