Caspase-mediated Apoptotic Effects of Ebenus boissieri Barbey Extracts on Human Cervical Cancer Cell Line HeLa

胱天蛋白酶介导的 Ebenus boissieri Barbey 提取物对人宫颈癌细胞系 HeLa 的凋亡作用

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作者:Ece Simsek, Nilufer Imir, Esra Arslan Aydemir, Ramazan Suleyman Gokturk, Erdem Yesilada, Kayahan Fiskin

Background

Ebenus boissieri Barbey is an Antalya, Turkey-endemic plant belonging to Fabaceae family. The aerial parts and the roots of E. boissieri Barbey were used in this study.

Conclusion

The effects of the extract on caspase-3, TNF-α, and IFN-γ levels mediate the plausible mechanism of apoptosis induction in HeLa. To the best of our knowledge, this is the first report indicating any pharmacological properties of E. Boissieri on HeLa cells. Summary: HeLa cell viability was reduced in dose-dependent manner for 72 h with an IC50 of approximate 28.03 μg/mL for aerial and 41.02 μg/mL for rootHeLa cells, exposure to the aerial extract led to 1.9, 3.8, 1.2, 2.4, and 3.45 fold induction of all caspases activities (-2, -3, -6, -8, and -9, respectively)Both 30 μg/mL of aerial and 45 μg/mL of root extracts allowed the production of anticancer cytokines (TNFalpha; IFNgamma) in HeLa cell culture supernatants. Abbreviations used: Tumor necrosis factor-alpha (TNF-α); Interferon gamma (IFN-γ); 3-(4, 5 dimethylthiazol-2-yl)-5-(3- carboxymethoxy-phenyl)-2-(4-sulfonyl)-2H-tetrazolium (MTS); Phosphate-Buffered Saline (PBS); Fetal Bovine Serum (FBS); para-Nitroanilin pNA; Enzyme-Linked ImmunoSorbent Assay (ELISA); Sodium Dodesyl sulphate -Polyacrilamide gel electrophoresis (SDS-PAGE); Tris-Buffered Saline (TBS); Hydocloric acid (HCl); Standart Error of Mean (SEM); National Cancer Institute (NCI); half maximal inhibitory concentration (IC50).

Methods

To determine the cytotoxic effect, cells were treated with various concentrations of extracts for 24, 48, and 72 h incubation periods. Cytotoxic effects were examined by Cell Titer 96 aqueous nonradioactive cell proliferation assay and the

Objective

In the present study, we have examined the apoptotic effects of hydroalcoholic extracts of E. boissieri Barbey in human cervical cancer cell line HeLa. Materials and

Results

According to the results, E. boissieri Barbey extract caused significant increase in caspase levels. Thus, we suggest that the extract induces cells to undergo apoptosis. Especially, there was a sharp induction in caspase-3 activity. Levels of both TNF-α and IFN-γ in extract-treated groups were significantly and dose dependently exalted as compared to their relative controls.

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