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

Öz

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.

Anahtar Kelimeler

• Verbascum inulifolium,breast cancer,antiproliferation,, apoptozis

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

Öz

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.

Anahtar Kelimeler

• Verbascum inulifolium,breast cancer,antiproliferation,apoptozis

Kaynakça

1. Elmore S. Apoptosis: a review of programmed cell death Toxicol Pathol 2007; Jun;35:495-516.
2. Vousden KH, Lu X. Live or let die: the cells response to p53. Nat Rev Cancer 2002;2:594- 604.
3. Ju J, Zhu AJ, Yuan, P. Progress in targeted therapy for breast cancer. Chron Dis Transl Med 2018;4:164-175.
4. Kintzios SE. Terrestrial plant-derived anticancer agents and plant species used in anticancer research. Crit rev plant sci 2006; 25:79-113.
5. Kayır S, et al. The in vivo effects of Verbascum speciosum on wound healing. S Afr J Bot 2018; 119;226-229.
6. Mihailović V, et al. Chemical profile, antioxidant activity and stability in stimulated gastrointestinal tract model system of three Verbascum species. Ind Crop Prod 2016; 89:141-151.
7. Kozan E, et al.The in vivo anthelmintic efficacy of some Verbascum species growing in Turkey. Exp parasitol 2011;129:211-214.
8. Dalar A, Guo Y, Konczak I. Phenolic composition and potential anti-inflammatory properties of Verbascum cheiranthifolium var. cheiranthifolium leaf. J of Herb Med 2014; 4:195-200.

9. Iqbal J, et al.. Plant-derived anticancer agents: A green anticancer approach. Asian Pac J Trop Biomed 2017;7:1129-1150.
10. Cragg GM, Newman DJ. Plants as a source of anti-cancer agents. J ethnopharmacol 2005;100:72-79.
11. Suntar I, et al. An ethnopharmacological studyon Verbascum species: fromconventionalwound healing usetoscientific verification. J ethnopharmacol 2010;132:408–413.
12. Dülger B, et al. Antimicrobial activity of three endemic Verbascum species. Pharm biol 2002; 40:587-589.
13. Grigore A, et al. Correlation between polyphenol content and anti-inflammatory activity of Verbascum phlomoides (mullein). Pharm biol 2013; 51:925-929.
14. Gürbüz I, et al. Anti-ulcerogenic activity of some plants used in folk medicine of Pinarbasi (Kayseri, Turkey). J ethnopharmacol 2005;101: 313-318.
15. Talib WH, Mahasneh AM. Antiproliferative activity of plant extracts used against cancer in traditional medicine. Sci pharm 2010; 78: 33.
16. Dimitrova P, et al. New iridoids from Verbascum nobile and their effect on lectin-induced T cell activation and proliferation. Food Chem Toxicol 2018; 111:605-615.
17. Zhao YL, et al. Isolation of chemical constituents from the aerial parts of Verbascum thapsus and their antiangiogenic and antiproliferative activities. Arch of pharm res 2011;34:703-707.
18. Chuang HC, et al. Cytotoxic effects of incense particles in relation to oxidative stress, the cell cycle and F-actin assembly. Toxicol lett 2013; 220: 229-237.