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

Year 2019, , 203 - 210, 31.05.2019

Yusuf Özay , Atila Yıldız , Celal Güven , İşıl Albeniz Leyla Türker Şener , Mufide Aydoğan Ahbab , İbrahim Bozgeyik Sevda Güzel Haydar Bağış Önder Yumrutaş

https://doi.org/10.31020/mutftd.532710

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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