İnsan Kolon Kanser Hücre Hattında (HT29) Sisplatin'in Apoptotik Etkisi

Abstract

Amaç: Çalışmamızda, sisplatinin 72 saatlik farklı dozlarda insan kolon kanseri olan HT29 hücre hattındaki etkilerinin araştırılması hedeflenmiştir. Gereç ve Yöntem: 5-200 μM' lık dozlar arasında sisplatin eklenen hücreler 72 saat süresince kültüre edilmiştir. EtBr-AO boyama tekniği apoptozun değerlendirilmesinde kullanılmıştır. BCL2L1, CASP3 ve CASP8 genlerinin mRNA ifade düzeyleri RTPCR yöntemi ile değerlendirilmiştir. Bulgular: Sisplatinin 72 saatte doza bağlı sitotoksik etki gösterdiği ve uygun apoptotik dozunun yine bu saat için 50μM olduğu tespit edilmiştir. Araştırılan üç genin mRNA düzeylerinin, 5μM sisplatin uygulandığında kontrole göre istatistiksel olarak anlamlı artış gösterdiği belirlenmiştir (p<0.05). 50μM sisplatin uygulamasının ise BCL2L1 mRNA düzeyinde kontrole göre değişiklik oluşturmadığı (p>0.05), ancak aynı doz ve sürede CASP3 mRNA düzeyinin 2.2 kat, CASP8 mRNA düzeyinin 3.0 kat arttığı bulunmuştur (p<0.05). Sonuç: Çalışmamızda insan kolon kanser hücrelerinde sisplatinin 72'inci saatte ve 50μM dozda en etkin değerde apoptozu uyardığı, sinyal iletim yolaklarından kaspaz kaskadını kullandığı ve antiapoptotik yolağı baskıladığı görülmüştür.

Anahtar Kelimeler: Apoptoz, HT29, Hücre kültürü, Kolon kanseri, Sisplatin

Keywords

• Apoptoz, HT29, Hücre kültürü, Kolon kanseri, Sisplatin

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