Acacia catechu ethanolic bark extract induces apoptosis in human oral squamous carcinoma cells

Abstract

Oral cancer is in approximately 30% of all cencers in India. This study was conducted to evaluate the cytotoxic activity of ethanolic extract of Acacia catechu bark (ACB) against human squamous cell carcinoma cell line-25 (SCC-25). Cytotoxic effect of ACB extract was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide assay. A. catechu extract was treated SCC-25 cells with 50 mu g/mL for 24h. Apoptosis markers such as capases-8 and 9, bcl-2, bax, and cytochrome c(Cyt-c) were done by RT-PCR. Morphological changes of ACB treated cells were evaluated using acridine orange/ethidium bromide (AO/EB) dual staining. Nuclear morphology and DNA fragmentation were evaluated using propidum iodide (PI) straining. Further, cell cycle analysis was performed using flow cytometry.A. catechu treatment caused cytotoxicity in SCC-25 cells with an IC50 of 52.09 mu g/mL. Apoptotic marker gene expressions were significantly increased on ACB tratment. Staining with AO/EB and PI shows membrane biebbing and nuclear membrane disortion, respectively, and it confirms the apoptosis induction in SCC-25 cells. These results suggest that ACB extract can be used as a modulating agent in oral squamous cell carcinoma.