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Antioxidant, anticancer and apoptotic effects of the Bupleurum chinense root extract in HO-8910 ovarian cancer cells.

PURPOSE
The purpose of this study was to evaluate the anticancer, apoptotic and antioxidant properties of Bupleurum chinense (B.C) root extract against human epithelial ovarian cancer cells (HO-8910) in vitro.


METHODS
MTT assay was used to evaluate the cell viability of HO-8910 cells after treatment with different B.C extract doses. Apoptotic and morphological effects induced by the extract were demonstrated by inverted phase contrast microscopy and fluorescence microscopy. The percentage of apoptotic cells was quantified by Annexin V/PI double staining assay. Flow cytometry using rhodamine-123 dye was used to measure disruption of mitochondrial membrane potential (Δψm). Gel electrophoresis was used to study the effects of the extract on DNA fragmentation. The antioxidant activity of the extract using 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazolin-6-sulphonic acid) (ABTS) radical scavenging assays was also evaluated.


RESULTS
The results showed that B.C extract could induce potent and dose-dependent cytotoxic effects on the HO-8910 cells as demonstrated by MTT assay. The extract also induced cell shrinkage, chromatin condensation and membrane blebbing which are the hallmark of apoptosis. The average proportion of Annexin V-staining positive cells (total apoptotic cells) significantly increased from 9.4% in control cells to 18.5, 28.2 and 50.5% in 20, 80 and 120 μg/ml B.C extract-treated cells respectively. Different doses of the extract (20, 80 and 120 μg/ml) after 48 hrs exposure led to a substantial increase in DNA fragmentation.The number of cells with disrupted Δψm increased from 6.6% in untreated (control cells) to 14.2, 42.1 and 73.4% in 20, 80 and 120 μg/ml in extract-treated cells, respectively


CONCLUSION
The anticancer effects of Bupleurum chinense extract were mediated through the induction of apoptosis, DNA fragmentation and disruption of mitochondrial membrane potential.