Potential Antioxidant and Cytotoxic Activities of Areca Nut (Areca Catechu Linn.) Extract in Human Oral Squamous Cell Carcinoma and Keratinocyte Cells

Abstract

Objectives: Areca (betel, Areca catechu) nuts have been reported to show high contents of phenolic and flavonoid compounds with associated antioxidant activity. This study was undertaken to assess the anticancer potential of areca nut extract on human squamous carcinoma (HSC)-3, HSC-2, and human keratinocyte (HaCat) cancer cell lines, and to elucidate the mechanism of its action. Methods:In this study, the areca nut extract was prepared in 96% ethanol and analyzed for the total phenolic and flavonoid content, 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, and 5-(3-carboxymethoxyphenyl)-2-(4,5-dimenthylthiazoly)-3-(4-sulfophenyl)tetrazolium inner salt assay. Results:The indicated total phenolic content of the areca nut extract was 80.3 mg tannic acid equivalent/g and total flavonoid content 238.5 mg catechin equivalent/g. The extract showed good scavenging activity against DPPH radical (efficient concentration 15.95±0.84 μg/ml). The extract showed higher toxicity on HSC-3 than HSC-2 cells, IC 50 was 164.06 μg/ml in HSC-3 versus 629.50 μg/ml in HSC-2. The extract had no cytotoxic effect on HaCat cells. Conclusions:The antioxidant activity of the areca nut extract might be associated with the presence of high content of phenolic and flavonoid compounds. The extract showed selective cytotoxicity against HSC-3 and HSC-2 cells but not on HaCat cells. Instead, the extract induced high rate of proliferation in HaCat cells.

Areca nut extract demonstrated apoptosis-inducing mechanism by increased caspase-3 activities on oral squamous cell carcinoma

“…This study is a novel or first study which clearly reveals the potential cytotoxicity effect and mechanism of action of areca nut in oral squamous cell lines. Our preliminary study showed that the areca nut has a high content of total phenolic and flavonoid compounds, and showed cytotoxicity activity against HSC-2 and HSC-3 cells 38 . In the present study, we found that areca nut extract can induce apoptosis in HSC-2 and HSC-3 cells with optimum time after 48 hours exposure.…”

“…The pellet was homogenized by pipetting, and the resuspended cells with the culture medium were ready to be used for experiment and cell counting with a hemocytometer. We had performed the cell viability assay previously to evaluate the percentage cytotoxicity and IC 50 of areca nut extract after treating the HSC-2 cells for 72 hours is 629.50 µg/mL while in HSC-3 cells is 164.06 µg/mL 38 . The protocol was approved by the Ethics Committee of the Faculty of Medicine, University of Indonesia no.…”