İki enginar türünden elde edilen bitkisel ekstraktların kolorektal kanser hücre dizisi üzerinde etkilerinin araştırılması
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Dosyalar
Tarih
2012-08-07
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Selçuk Üniversitesi Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Geleneksel olarak bitkilerin kullanımı ülkemizde uzun zamandan beri yaygın olarak tercih edilmekte, etnobotanik çalışmaların modern tıbbi çalışmalara olan katkısı her geçen gün artmakta ve değerli bilgiler elde edilmektedir. Bu tez çalışması ile ülkemizde doğal olarak yetişen (Cynara syriaca) ve kültüre alınan (Cynara cardunculus) enginar türlerinden elde edilen bitkisel ekstraktların kolorektal kanser hücre hattı DLD-1 üzerine olan apoptotik ve sitotoksik etkisinin araştırılması amaçlanmıştır. Bu amaçla kültür ve yabani enginarın vegetatif kısımlarından ve yenen kısmı olan resaptakulumdan hekzan kullanılarak ekstraklar elde edilmiştir. Bu ekstraktlar ve apigenin, MTT testi için DLD -1 hücreleri üzerine beş farklı dozda (0,1-1 mg/ml) ve üç farklı zaman aralığında (24-48-72 saat) uygulanmış, doza ve zamana bağlı değişen sitotoksik aktiviteleri belirlenmiştir. Elde edilen MTT verileri tek yönlü ANOVA ve Dunnett testleri ile istatistiksel olarak değerlendirilmiş, anlamlı farklılıklar gözlenmiştir. Hücre ölümüne neden olan gerçek sebebin ne olduğunu belirlemek amacıyla, her ekstrakt için hesaplanan IC50 dozu hücre hatları üzerine uygulanmış ve bu hatlardan total RNA ve DNA izolasyonu gerçekleştirilmiştir. İzole edilen DNA'lar (%1. 5) agaroz jele yüklenerek fragmantasyon analizi yapılmış, izole edilen RNA'lardan cDNA sentezi ve sırasıyla p53, p21, Bcl-2, Bax gen bölgelerinin amplifikasyonları yapılmıştır. Gen ekspresyon düzeyleri Image J programı kullanılarak ölçülmüştür. Sonuçlar bu gen bölgelerinin ifade seviyeleri ile ilişkilendirilmiştir.
Traditionally, the using of plants has been commonly preferred for a long time and the conclusions to modern medical research from ethno botanical studies have rated day to day and very important information have been obtained from them. By this master thesis, it was aimed to determine apoptotic and cytotoxic activity of plant extracts obtaining from naturally growing (Cynara syriaca) in our country and cultivated (Cynara cardunculus) Cynara species against to DLD1 colorectal cancer cells. By this aim, the extracts belonging to wild and cultivated Cynara species were obtained from their vegetative parts and receptaculum by using hexane. These extracts besides apigenin for MTT test were applied with five different dose (0,1-1 mg/ml) and by three time interval (24-48 and 72nd hours) upon DLD1 cancer cells and cytotoxic activities changed based on dose and time dependent manner were determined. The data obtaining from MTT test were evaluated statistically via One Way ANOVA and Dunnett test and meaningful differences from them were observed. After IC50 dose counted for each extract were firstly superimposed on cell lines and then total DNA and RNA were isolated from these lines for determination of the real reason causing cell deaths. Thereby the isolated DNAs were loaded to (1.5 %) agarose gel, fragmentation analyses were carried out. From isolated RNAs were synthesed cDNA and the amplifications of p53, p21, Bcl-2 and Bax gene regions were done respectively. The levels of gene expression were measured by using Image J programme. Results were associated with the expression levels of these gene regions.
Traditionally, the using of plants has been commonly preferred for a long time and the conclusions to modern medical research from ethno botanical studies have rated day to day and very important information have been obtained from them. By this master thesis, it was aimed to determine apoptotic and cytotoxic activity of plant extracts obtaining from naturally growing (Cynara syriaca) in our country and cultivated (Cynara cardunculus) Cynara species against to DLD1 colorectal cancer cells. By this aim, the extracts belonging to wild and cultivated Cynara species were obtained from their vegetative parts and receptaculum by using hexane. These extracts besides apigenin for MTT test were applied with five different dose (0,1-1 mg/ml) and by three time interval (24-48 and 72nd hours) upon DLD1 cancer cells and cytotoxic activities changed based on dose and time dependent manner were determined. The data obtaining from MTT test were evaluated statistically via One Way ANOVA and Dunnett test and meaningful differences from them were observed. After IC50 dose counted for each extract were firstly superimposed on cell lines and then total DNA and RNA were isolated from these lines for determination of the real reason causing cell deaths. Thereby the isolated DNAs were loaded to (1.5 %) agarose gel, fragmentation analyses were carried out. From isolated RNAs were synthesed cDNA and the amplifications of p53, p21, Bcl-2 and Bax gene regions were done respectively. The levels of gene expression were measured by using Image J programme. Results were associated with the expression levels of these gene regions.
Açıklama
Anahtar Kelimeler
DLD-1, CDNA, DNA fragmantasyonu, Enginar, Kolorektal kanser, MTT, RT-PCR, Artichoke, Colorectal cancer, DNA fragmentation
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Şimşek, E. N. (2012). İki enginar türünden elde edilen bitkisel ekstraktların kolorektal kanser hücre dizisi üzerinde etkilerinin araştırılması. Selçuk Üniversitesi, Yayımlanmış yüksek lisans tezi, Konya.
