MCF7 meme kanseri hücreleri üzerine Verbascum inulifolium’un antiproliferatif ve apoptoz indükleme aktivitelerinin değerlendirilmesi

Abstract

Verbascum cinsi halk tıbbında geleneksel olarak kullanılan ve anti-enflamatuvar, antioksidan ve antibakteriyal aktiviteleri bilinen bir cinstir. Verbascum inulifolium türü Türkiye’de yetişen endemik bir türdür. Bu çalışmanın amacı V. inulifolium türünün toprak üstü kısımlarından elde edilen metanol ekstresinin insan meme kanseri hücre hattı (MCF7) üzerine antiproliferatif ve apoptotik etkilerinin değerlendirilmesidir. Ekstrenin farklı konsantrasyonlarının (1, 5, 25 ve 125µg/ml) hücre canlılığı ve proliferasyonu üzerine etkisi MTT ve Xcellange yöntemi ile test edilmiştir. Daha sonra hücreler Annexine V/propodium iodür (PI) ile boyanmış ve apoptozun safhaları FACS cell counter metodu ile belirlenmiştir. Ayrıca apoptotik ve anti-apoptotik protein ekspresyon düzeyleri western blot metodu ile belirlenmiştir. Son olarak phallodine boyama metodu ile hücrelerde aktin flamentleri boyanmış ve floresan mikroskobunda incelenmiştir. MCF-7 hücre canlılığı ve proliferasyonun doz artışına bağlı olarak azaldığı belirlenmiştir. Ancak apoptozun erken veya geç dönemlerinde MCF-7 hücre sayılarında bir değişim gözlenmemiştir. P53, Bcl-2 ve kaspaz-3 protein ekspresyon düzeylerinde anlamlı bir değişim gözlenmemiştir. Ayrıca aktin flamentlerinin yoğunluğu sadece 125 mg/ml dozda hafif azalmıştır. Sonuç olarak V. inulifolium türünün methanol ekstresi MCF-7 hücreleri üzerine anti-proliferatif etki göstermiştir, fakat sonuçlar bu etkinin apoptotik yolakla ilişkili olmadığı göstermiştir.

Keywords

• Verbascum inulifolium,breast cancer,antiproliferation,, apoptozis

Antiproliferative and apoptosis inducing effect of Verbascum inulifolium on MCF-7 breast cancer cell line

Abstract

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.

Keywords

• Verbascum inulifolium,breast cancer,antiproliferation,apoptozis

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