Androctonus crassıcauda (Olıver 1807) akrep ham zehri uygulanmış kolon kanseri hücre hattı (HT-29) üzerindeki gen expresyon düzeyinin qpcr yöntemi ile belirlenmesi
[ X ]
Tarih
2024
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Kırıkkale Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Kolon kanseri, Türkiye'de en yaygın kanser türlerinden biridir. Kolon kanseri, erken teşhis edilemediğinde ileri evrelerde tespit edildiği durumlarda yüksek ölüm oranlarına sahiptir. Bazı akreplerin zehirleri başta Küba ve Çin olmak üzere geleneksel tıpta kanser tedavisinde kullanılmaktadır. Ancak bu zehirlerin kanser tedavisindeki potansiyeli hakkında çok az bilimsel kanıt bulunmaktadır. Bu çalışmada, Androctonus crassicauda (Oliver 1807) akrebinden elde edilen ham zehrin Kolon Kanseri Hücre Hattı (HT-29) ve İnsan Embriyonik Böbrek Epitel Hücre Hattı (HEK-293) üzerinde in vitro koşullarda uygulanması sonucunda kanser ve normal epitel hücrelerinin üzerindeki etkileri araştırılmıştır. Akrep zehrinin farklı konsantrasyonları HT-29 kolon kanseri ve HEK-293 hücre hatlarının üzerine uygulandı (MTT hücre canlılığı testi ile HT-29 hücre hattında 40,54 mg/ml ve HEK-293 hücre hattında 57,3 mg/ml olarak IC50 (İnhibitör Konsantrasyon 50) değeri belirlendi). Belirlenen IC50 dozunda akrep zehri 24 saat süre ile HT-29 hücre hattına uygulanarak apoptoz mekanizması qPCR yöntemi kullanılarak değerlendirildi. Uygulanan dozda akrep zehrinin, apoptozun hangi yolağını indüklediğini belirlemek için Bax ve Bcl-2 genlerinin ekspresyon kantifikasyonu GAPDH geni referans olarak kullanılıp kontrol grubuna göre normalize edildi. Relatif kantifikasyon "2-??CT Yöntemi" kullanılarak yapılan hesaplamanın sonucuna göre Bax proteini kontrol grubu ile kıyaslandığında HT-29 hücre hattında yaklaşık 0,25 kat azaldığı ve HEK-293 hücre hattında yaklaşık 5 kat arttığı gözlemlendi. Bcl-2 gen ekspresyon düzeyinin kontrol grubuna göre HT-29 hücre hattında yaklaşık olarak 0,5 kat oranında azaldığı ve HEK-293 hücre hattında 2,5 kat arttığı gözlemlendi. Bu çalışma, A. crassicauda zehrinin kolon kanseri tedavisinde umut verici doğal bir ajan olabileceği konusunda önemli veriler içermektedir. Anahtar Kelimeler: Androctonus crassicauda, zehir, HT-29 kanser hücre hattı, Apoptoz, Sitotoksisite, qPCR
Colon cancer is one of the most common types of cancer in Türkiye. If detected in advanced stages or not diagnosed early, colon cancer has high mortality rates. The venom of some scorpions is used in traditional medicine to treat cancer, especially in Cuba and China. However, there is little scientific evidence about the potential of these poisons in treating cancer. In this study, the effects of crude venom obtained from the scorpion Androctonus crassicauda (Oliver 1807) on cancer and normal epithelial cells have been investigated following its in vitro application on Colon Cancer Cell Line (HT-29) and Human Embryonic Kidney Epithelial Cell Line (HEK-293). Different concentrations of scorpion venom have been applied on HT-29 colon cancer and HEK-293 cell lines by using MTT cell viability test. IC50 value (Inhibitor Concentration 50) has been determined as 40.54 mg/ml in HT-29 cell line and 57.3 mg/ml in HEK-293 cell line. Scorpion venom at the determined IC50 dose has been applied to the HT-29 cell line for 24 hours and the apoptosis mechanism has been evaluated using the qPCR method. To determine which pathway of the apoptosis the applied dose of scorpion venom induces, the expression quantification of Bax and Bcl-2 genes has been normalized compared to the control group, using the GAPDH gene as a reference. According to the results of the calculation by the relative quantification "2-??CT method", it has been observed that Bax protein has decreased approximately 0.25-fold in the HT-29 cell line and increased approximately 5-fold in the HEK-293 cell line compared to the control group. It has also been found out that the Bcl-2 gene expression level has decreased approximately 0.5-fold in the HT-29 cell line and increased 2.5-fold in the HEK-293 cell line compared to the control group. This study contains important data on A. crassicauda venom potentially being a promising natural agent in the treatment of colon cancer. Keywords: Androctonus crassicauda, venom, HT-29 cancer cell line, Apoptosis, Cytotoxicity, qPCR
Colon cancer is one of the most common types of cancer in Türkiye. If detected in advanced stages or not diagnosed early, colon cancer has high mortality rates. The venom of some scorpions is used in traditional medicine to treat cancer, especially in Cuba and China. However, there is little scientific evidence about the potential of these poisons in treating cancer. In this study, the effects of crude venom obtained from the scorpion Androctonus crassicauda (Oliver 1807) on cancer and normal epithelial cells have been investigated following its in vitro application on Colon Cancer Cell Line (HT-29) and Human Embryonic Kidney Epithelial Cell Line (HEK-293). Different concentrations of scorpion venom have been applied on HT-29 colon cancer and HEK-293 cell lines by using MTT cell viability test. IC50 value (Inhibitor Concentration 50) has been determined as 40.54 mg/ml in HT-29 cell line and 57.3 mg/ml in HEK-293 cell line. Scorpion venom at the determined IC50 dose has been applied to the HT-29 cell line for 24 hours and the apoptosis mechanism has been evaluated using the qPCR method. To determine which pathway of the apoptosis the applied dose of scorpion venom induces, the expression quantification of Bax and Bcl-2 genes has been normalized compared to the control group, using the GAPDH gene as a reference. According to the results of the calculation by the relative quantification "2-??CT method", it has been observed that Bax protein has decreased approximately 0.25-fold in the HT-29 cell line and increased approximately 5-fold in the HEK-293 cell line compared to the control group. It has also been found out that the Bcl-2 gene expression level has decreased approximately 0.5-fold in the HT-29 cell line and increased 2.5-fold in the HEK-293 cell line compared to the control group. This study contains important data on A. crassicauda venom potentially being a promising natural agent in the treatment of colon cancer. Keywords: Androctonus crassicauda, venom, HT-29 cancer cell line, Apoptosis, Cytotoxicity, qPCR
Açıklama
Fen Bilimleri Enstitüsü, Biyoloji Ana Bilim Dalı, Biyoloji Bilim Dalı
Anahtar Kelimeler
Biyoloji, Biology
