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Article Details

  • Article Code : FIRAT-AKADEMI-8992-4327
  • Article Type : Araştırma Makalesi
  • Publication Number : 4B0014
  • Page Number : 64-71
  • Doi : 10.12739/WSA.2017.12.4.4B0014
  • Abstract Reading : 680
  • Download : 147
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Issue Details

  • Year : 2017
  • Volume : 12
  • Issue : 4
  • Number of Articles Published : 4
  • Published Date : 1.10.2017

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Life Sciences

Serial Number : 4B
ISSN No. : 1308-7347
Release Interval (in a Year) : 4 Issues

LAETIPORUS SULPHUREUS (BULL.) MURRILL’UN SİTOTOKSİTE, APOPTİK VE NEKROTİK ETKİLERİ

Aydın KESKİN1 , Perihan GÜLER2 , Mustafa TÜRK3

Çalışmada, Laetiporus sulphureus’un sitotoksite, apoptik ve nekrotik etkileri incelenmiştir. Laboratuvardaki basidiokarplardan alınan parçalar malt ekstrakt agarda doku kültürü yöntemiyle geliştirildi. Primer ve sekonder miseller 27°C’de, karanlıkta, 10 gün süreyle inkübe edilerek elde edildi. Sekonder misellerden 3’er pelet nutrientbroth’da 27°C’de, 140 rpm’de 7 günde gelişimini tamamladı. Numuneler soxlet cihazında 80°C, %70’lik etil alkolde 1 saat ekstraksiyon edilerek sitoksite, apoptik ve nekrotik çalışmalar için hazırlandı. Sitoksite çalışmalarında, ekstrakt MCF-7 ve L929 Fibroblast hücrelere örnek konsantrasyonları 0.5mg/ml, 0.25mg/ml, 0.125mg/ml,0.0625mg/ml, 0.03125mg/ml olarak uygulandı. WST-1 solüsyonu eklenerek inkübasyon sonucunda 440nm dalga boyunda mikroplate okuyucuda absorbans ölçümü yapıldı. İkili boyama deneyi ile aynı hücre hattında apoptoz ve nekroz belirlendi. İkili boyama solüsyonu hazırlanarak MCF-7 ve L929 Fibroblast hücrelere aynı konsantrasyonlarda uygulandı. İnkübasyon sonucunda florasanataçmanlı mikroskop FITC ve DAPI filtresinde 20X büyütmede ölçüm yapıldı. Kullanılan 0.25 dozajda MCF-7 hücre hatlarında %10 sitotoksitite gözlenmiş. Kullanılan tüm dozajlarda L929 Fibroblast hücre hatlarında canlılık oranları artmıştır. Örneklerde apoptik ve nekrotik etki gözlenmiştir.

Keywords
Laetiporus sulphureus, Kanser, Sitotoksite, Apoptik Etki, Nekrotik Etki,

THE CYTOTOXIC, APOPTOTIC AND NECROTIC EFFECCTS OF LAETIPORUS SULPHUREUS (BULL.) MURRILL

Aydın KESKİN1 , Perihan GÜLER2 , Mustafa TÜRK3

In the study, cytotoxicity, apoptotic, necrotic effects of laetiporus sulphureus were investigated. Taken parts from the basidiocarps in the laboratory were developed with tissue culture on malt extract agar. The primary and the secondary mycelium were obtained by incubation at 27°C in the dark for 10 d. Three pellets were taken from the secondary mycelium and were completed their growth in 7 d at 140 rpm at 27°C in nutrient broth. The samples were prepared for cytotoxic, apoptotic, necrotic, studies by extracting at 80°C and 1 h in ethyl alcohol at soxlet device. In cytotoxic studies, sample concentrations for extract MCF-7 and L929 fibroblast cells were applied as 0.5mg/ml, 0.25mg/ml, 0.125mg/ml, 0.0625mg/ml, 0.03125mg/ml respectively. The WST-1 solution was added and the absorbance was measured in a microplate reader at a wavelength of 440 nm at the end of the incubation. Apoptosis and necrosis were detected in same cell line as the double staining experiment. Binary staining solution was prepared and MCF-7 and L929 fibroblast fibroblast cells were applied at the same concentrations. As a result of incubation, the measurement was done at fluorescense microscopy at 20X magnification in FITC and DAPI filters. Ten percent toxicity was observed at 0.25mg/ml dosage used in MCF-7 cell line, At all dosages used, viability rates were increased in L929 Fibroblast cell lines. Apoptotic and necrotic effects were observed in the samples.

Keywords
Laetiporus sulphureus, Cancer, Cytotoxicity, Apoptotic Effects, Necrotic Effects,

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Authors

Aydın KESKİN (1) (Corresponding Author)

Kırıkkale University
dr.emelkeskin@windowslive.com | 0000-0000-0000-0065

Perihan GÜLER (2)

Kırıkkale Üniversitesi
perihanguler71@gmail.com

Mustafa TÜRK (3)

mtrk.35@gmail.com | 0000-0000-0000-0064

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References
[1] Adanacıoğlu, N., Yıldız, Ü., Oğur, E., Aykas, L., Tan, A. ve Taylan, T., (2016). Türkiye Makromantarlar Genetik Kaynaklar 1. Ege Bölgesi. Anadolu, J. of Arı, 26(1), 2016, 46-61.