Apoptosis induction by polygonum minus is related to antioxidant capacity, alterations in expression of apoptotic-related genes, and S-phase cell cycle arrest in HepG2 cell line

• Published in: BioMed Research International
• Main Author: Mohd Ghazali M.A.; Al-Naqeb G.; Krishnan Selvarajan K.; Hazizul Hasan M.; Adam A.
• Format: Article
• Language: English
• Published: Hindawi Publishing Corporation 2014
• Online Access: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84901756283&doi=10.1155%2f2014%2f539607&partnerID=40&md5=6aec5eb29b0207bfa31c4ba60ff4c887

Polygonum minus (Polygonaceae) is a medicinal herb distributed throughout eastern Asia. The present study investigated antiproliferative effect of P. minus and its possible mechanisms. Four extracts (petroleum ether, methanol, ethyl acetate, and water) were prepared by cold maceration. Extracts were subjected to phytochemical screening, antioxidant, and antiproliferative assays; the most bioactive was fractionated using vacuum liquid chromatography into seven fractions (F1-F7). Antioxidant activity was measured via total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) assays. Antiproliferative activity was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Most active fraction was tested for apoptosis induction and cell cycle arrest in HepG2 cells using flow cytometry and confocal microscopy. Apoptotic-related gene expression was studied by RT-PCR. Ethyl acetate extract was bioactive in initial assays. Its fraction, F7, exhibited highest antioxidant capacity (TPC; 113.16 ± 6.2 mg GAE/g extract, DPPH; EC 50: 30.5 ± 3.2 g/mL, FRAP; 1169 ± 20.3 mol Fe (II)/mg extract) and selective antiproliferative effect (IC 50: 25.75 ± 1.5 g/mL). F7 induced apoptosis in concentration- and time-dependent manner and caused cell cycle arrest at S-phase. Upregulation of proapoptotic genes (Bax, p53, and caspase-3) and downregulation of antiapoptotic gene, Bcl-2, were observed. In conclusion, F7 was antiproliferative to HepG2 cells by inducing apoptosis, cell cycle arrest, and via antioxidative effects. © 2014 Mohd Alfazari Mohd Ghazali et al.