ECHINOCOCCUS GRANULOSUS SENSU STRICTO KAYNAKLI HİDATİK KİST SIVISI UYGULAMASININ CACO-2 HÜCRE HATTINDA EPİTELYAL-MEZENKİMAL GEÇİŞ VE APOPTOZ ÜZERİNE ETKİSİ

Year 2022, , 1 - 8, 30.04.2022

İpek Baysal Serra Örsten

https://doi.org/10.54962/ankemderg.1107804

Abstract

Kistik ekinokokkoz (KE), Echinococcus granulosus sensu lato'nun larva formunun neden olduğu zoonotik bir enfeksiyondur. Yapılan çalışmalar E. granulosus enfeksiyonu ile kanser arasında doğrudan ve/veya dolaylı bir ilişki olduğu öne sürmüş; ancak, elde edilen sonuçlar farklı hücre kültürü ve/veya hayvan modellerinde hem anti-kanserojen hem de kanserojen etkisi olabileceğini göstermiştir. İnsan kolorektal adenokarsinom (Caco-2) hücrelerine etkisi daha önce değerlendirilmemiştir. Bu çalışmanın amacı, hidatik kist sıvısı uygulamasının bazı apoptotik genlerin (BCL-2, p53 ve BAX) ve epitelyal-mezenkimal geçiş (EMT) genlerinin (e-kaderin ve vimentin) ekspresyonu ve hücre proliferasyonu üzerine etkisini değerlendirerek Caco-2 hücre hattında olası anti-kanserojen veya kanserojen etkisini moleküler düzeyde aydınlatmaktır. Hidatik kist sıvısı uygulamasının sonrasında hücre proliferasyonu, apoptotik genler ve EMT gen ekspresyonu üzerindeki etkisini değerlendirmek için hücre proliferasyon analizi (XTT ile) ve gerçek zamanlı polimeraz zincir reaksiyonu (RT-PCR) yapılmıştır. Uygulama sonrasında uygulama dozu ile orantılı olarak hücre proliferasyonunda istatistiksel olarak anlamlı bir artış tespit edilmiştir. BAX ve p53 gen ifadelerinde doza bağlı azalma ve BCL-2 gen ifadesinde artış tespit edilmiştir. Bunun yanı sıra hücre kültüründe EMT gözlenmiş ve e-kaderin (CDH1) ve vimentin ekspresyonları ile moleküler düzeyde doğrulanmıştır. Bu çalışma ile hidatik kist sıvısının, Caco-2 hücre hattına uygulanması hücre proliferasyonunu doğrudan arttırdığı ve Caco-2 hücre hattının apoptoza karşı çok daha dirençli ve metastatik hale gelmesine neden olduğu tespit edilmiştir. Bu çalışma, ilk kez Caco-2 hücre hattından apoptotik yolakta hidatik kist sıvının olası mekanizmasına ışık tutmaktadır.

Keywords

apoptoz, Echinococcus granulosus, epitelyal-mezenkimal geçiş, kanser, kist hidatik sıvısı

The Effect of Echinococcus granulosus sensu stricto-Derived Hydatid Cyst Fluid Application on Epithelial-Mesenchymal Transition and Apoptosis on Caco-2 Cell Line

Abstract

Cystic echinococcosis (CE) is a zoonotic infection caused by the larval form of Echinococcus granulosus sensu lato. Studies suggested a direct and/or indirect relationship between E. granulosus infection and cancer; however, results showed both anti-carcinogenic and carcinogenic effects in different cell culture and/or animal models. Its effect on human colorectal adenocarcinoma (Caco-2) cells has not been previously evaluated. The aim of this study was elucidate the molecular mechanism underlying the possible anti-carcinogenic or carcinogenic effect of hydatid cyst fluid on Caco-2 cell line at the molecular level via evaluating the effect of its application on cell proliferation and expression of some apoptotic genes (BCL-2, p53 and BAX) and epithelial-mesenchymal transition (EMT) genes (e-cadherin and vimentin). Cell proliferation analysis (using XTT) and real-time polymerase chain reaction (RT-PCR) were performed to evaluate the effect of hydatid cyst fluid on cell proliferation and apoptotic and EMT gene expression, respectively. After the application, a statistically significant increase in cell proliferation was detected correlated with the application dose. A dose-dependent decrease in BAX and p53 gene expressions and an increase in BCL-2 gene expression were determined. EMT is also observed and confirmed via e-cadherin (CDH1) and vimentin expression levels. With this study, the application of hydatid cyst fluid to Caco-2 cells directly increases cell proliferation. Thus, it caused the Caco-2 cell line to become much more resistant to apoptosis and prone to metastasis. This study sheds light on the possible mechanism of hydatid cyst fluid in the apoptotic pathway from the Caco-2 cell line for the first time.

Keywords

apoptosis, cancer, Echinococcus granulosus, epithelial-mesenchymal transition, hydatid cyst fluid

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