Open Peer Review RESEARCH ARTICLE Areca nut extract demonstrated apoptosis-inducing mechanism by increased caspase-3 activities on oral squamous cell carcinoma [version 5; peer review: 2 approved] Liza Meutia Sari , Gus Permana Subita , Elza Ibrahim Auerkari 3 Oral Medicine Department, Faculty of Dentistry, University of Syiah Kuala, Banda Aceh, 23111, Indonesia Oral Medicine Department, Faculty of Dentistry, University of Indonesia, Jakarta, 10430, Indonesia Oral Biology Department, Faculty of Dentistry, University of Indonesia, Jakarta, 10430, Indonesia Abstract Oral squamous cell carcinoma is a neoplasm of keratinocyte Background: cells of oral mucosa epithelium that can potentially spread through lymphatic tissue or blood vessel. Although areca nut is one of the plants with a risk of inducing that cancer, areca nut is believed to have high antioxidant properties. Due to the current interest in the apoptosis effects from areca nut for oral cancer treatment, we investigated its ability to induce apoptosis and caspase-3 activity in oral cancer cell lines: HSC-2 and HSC-3. We examined the effect of areca nut on apoptosis and Methods: caspase-3 activity in HSC-2 and HSC-3 cells. Flow cytometry was conducted for the quantification of the cells that were apoptotic and expressing the caspase-3 enzyme for 24 and 48 hours. Areca nut induced a significant increase (p<0.01) in late apoptosis Results: of HSC-2 cells and mostly occurred over 48 hours. The study also found that in HSC-3, there were significant increases (p<0.01) the percentage of cells in early apoptosis after 24 hours and late apoptosis at 48 hours. Caspase-3 activity increased after 24 and 48 hours of areca nut exposure in both cells. The study showed that areca nut could be considered as a Conclusions: potential anticancer agent through its capability in inducing a caspase-dependent apoptosis. Keywords Areca nut, oral cancer, apoptosis, caspase-3 1 2 3 1 2 3 Reviewer Status Invited Reviewers version 5 published 01 Jul 2019 version 4 published 12 Feb 2019 version 3 published 07 Dec 2018 version 2 published 26 Oct 2018 version 1 published 11 Jun 2018 1 2 report report report report report report , Padjadjaran University, Irna Sufiawati Bandung, Indonesia , Solachuddin Jauhari Arief Ichwan International Islamic University Malaysia, 1 11 Jun 2018, :723 ( First published: 7 ) https://doi.org/10.12688/f1000research.14856.1 26 Oct 2018, :723 ( Second version: 7 ) https://doi.org/10.12688/f1000research.14856.2 07 Dec 2018, :723 ( Third version: 7 ) https://doi.org/10.12688/f1000research.14856.3 12 Feb 2019, :723 ( Fourth version: 7 ) https://doi.org/10.12688/f1000research.14856.4 01 Jul 2019, :723 ( Latest published: 7 ) https://doi.org/10.12688/f1000research.14856.5 v5 Page 1 of 34 F1000Research 2019, 7:723 Last updated: 30 JUL 2019