IADR Abstract Archives
Apoptotic Induction of Rhinacanthin-C Extracted From Rhinacanthus Nasutus on OSCC
Objectives: To study the apoptotic induction of Rhinacanthin-C extracted from Rhinacanthus nasutus on oral squamous cell carcinoma (OSCC) by detecting the activation of caspase-3 and the exposure of phosphatidylserine for further development as a novel anticancer treatment or a co-treatment.
Methods: The caspase-3 activation by Rhinacanthin-C on OSCC was determined by colorimetric assay using a specific peptide labeled with chromophore pNA. The detection of phosphatidylserine on the outer leaflet of the apoptotic cell membrane was monitored by flow cytometry using Annexin V-FITC and propidium iodide staining procedure. The statistical analysis was performed using SPSS software version 18. All experiments were repeated at least three times. The statistical differences among the groups were analyzed using Kruskal Wallis nonparametric test (p-value < 0.05).
Results: The incubation of Rhinacanthin-C on HSC4 (human oral squamous cell carcinoma) cells induced the activation of caspase-3 activity at all tested concentrations (1, 5, and 10 μM) with significant increases at 5 and 10 μM. Fifty μM of 5-Fluorouracil or 5-FU was used as a positive control. Based on the detection of the phosphatidylserine by flow cytometry, Rhinancanthin-C significantly increased the number of apoptotic cells in all tested concentrations (1, 5, and 10 μM) in a dose-dependent manner.
Conclusions: The apoptotic induction of Rhinacanthin-C extracted from Rhinacanthus nasutus was confirmed by the activation of caspase-3 and an increase in the exposure of phosphatidylserine.
Methods: The caspase-3 activation by Rhinacanthin-C on OSCC was determined by colorimetric assay using a specific peptide labeled with chromophore pNA. The detection of phosphatidylserine on the outer leaflet of the apoptotic cell membrane was monitored by flow cytometry using Annexin V-FITC and propidium iodide staining procedure. The statistical analysis was performed using SPSS software version 18. All experiments were repeated at least three times. The statistical differences among the groups were analyzed using Kruskal Wallis nonparametric test (p-value < 0.05).
Results: The incubation of Rhinacanthin-C on HSC4 (human oral squamous cell carcinoma) cells induced the activation of caspase-3 activity at all tested concentrations (1, 5, and 10 μM) with significant increases at 5 and 10 μM. Fifty μM of 5-Fluorouracil or 5-FU was used as a positive control. Based on the detection of the phosphatidylserine by flow cytometry, Rhinancanthin-C significantly increased the number of apoptotic cells in all tested concentrations (1, 5, and 10 μM) in a dose-dependent manner.
Conclusions: The apoptotic induction of Rhinacanthin-C extracted from Rhinacanthus nasutus was confirmed by the activation of caspase-3 and an increase in the exposure of phosphatidylserine.