Özet:

The genus Verbascum traditionally used in folk medicine is known with some activities including anti-inflammatory, antioxidant and antibacterial. Verbascum inulifolium is an endemic species growing in Turkey. The aim of the present study was to evaluate anti-proliferative and apoptotic effects of methanol extract obtained from aerial parts of V. inulifolium on human breast cancer cell line (MCF). Effects of different concentration of the extract (1, 5, 25 and 125μg/ml) were tested on cell viability and proliferation using MTT and Xcelligence assays. Then cells stained with Annexine V/propodium iodine (PI) and apoptosis phases were determined using FACS cell counter method. Moreover, the western-blot method was performed to determine apoptotic and anti-apoptotic protein expression levels. Finally, cell actin filaments stained using phallodine stain method and examined with fluorescence microscope. Cell viability and proliferation levels were reduced according to increasing concentrations of the extract. However, there were no differences observed in the number of MC-7 cells during early and late phases of apoptosis. There were no significantly differences observed in P53, Bcl-2, and caspase-3 protein expression levels. Actin filament density was slightly reduced only at the concentration of 125 mg/ml. As a result, methanol extract of V. inulifolium showed anti-proliferative effect, but results indicated that this effect was not related to apoptotic pathway.

Anahtar Kelimeler:

Özet:

The genus Verbascum traditionally used in folk medicine is known with some activities including anti-inflammatory, antioxidant and antibacterial. Verbascum inulifolium is an endemic species growing in Turkey. The aim of the present study was to evaluate anti-proliferative and apoptotic effects of methanol extract obtained from aerial parts of V. inulifolium on human breast cancer cell line (MCF). Effects of different concentration of the extract (1, 5, 25 and 125µg/ml ) were tested on cell viability and proliferation using MTT and Xcelligence assays. Then cells stained with Annexine V/propodium iodür (PI) and apoptosis phases were determined using FACS cell counter method. Moreover, western-blot method was performed to determine apoptotic and anti-apoptotic protein expression levels. Finally, cell actin filaments stained using phallodine stain method and examined with fluorescence microscope. Cell viability and proliferation levels were reduced according to increasing concentrations of the extract. However, there were no differences observed in number of MC-7 cell during early and late phases of apoptosis. There were no significantly differences observed in P53, Bcl-2, and caspase-3 protein expression levels. Actin filament density was slightly reduced only at the concentration of 125 mg/ml. As a result, methanol extract of V. inulifolium showed anti-proliferative effect, but results indicated that this effect was not related to apoptotic pathway.