Berberis hispanica alkaloids extract induced cell death and apoptosis in human laryngeal cancer cells Hep-2

Abstract Berberis (Barberry) species contain an important amount of alkaloids and are commonly used in traditional medicine. Alkaloids are preponderantly characterized by their numerous properties including anticancer activity. The aims of the present study were to investigate the effect and the mechanism of action of the Algerian Berberis hispanica alkaloids extract (BHAE) on Hep-2 laryngeal cancer cells. In this purpose, total alkaloids were extracted from the roots bark of the plant, then, were submitted to HPLC analysis. In order to determine the cells inhibition concentration (IC50), we used the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cells were enumerated using Trypan blue exclusion assay and viability by cell cycle analysis. The morphology was assessed by coloration with May Grunwald Giemsa (MGG). The evaluation of such stress markers as malondialdehyde (MDA), cytochrome c, p38MAPK, NF-kB, Akt, Erk 1/ 2 and p53 was assessed by colorimetric assays and western blotting. Our results demonstrated that BHAE contains Berberine, an isoquinoline plant alkaloid. The IC50 was determined at 75 μg/mL and the treatment of Hep-2 cells with this dose showed anti-proliferative effect and increased SubG0 cell population (cells undergoing apoptosis) in treated cells (15.4%). Further, the coloration of fixed cells showed that BHAE induced cells density and morphological changes. Moreover, our results showed that the treatment induced elevation of MDA, cytochrome c, p38MAPK and NF-kB level, but did not affect Akt and Erk 1/2 rates. We have also observed that the treatment with BHAE enhanced the expression of p53. Taken together, our results suggest that the BHAE triggered cell death by activation of apoptosis through ROS induction.