Year 2020, Volume 23 , Issue 5, Pages 1125 - 1134 2020-10-31

Kapsaisinin H2452 Mezotelyoma Hücre Hattından Seçilen Klon larda Sitotoksisitesinin ve Kaspaz-3 Üzerine Etkilerinin Belirlenmesi
Determination of the Cytotoxicity of Capsaicin and its Effects on Caspase-3 in the Clones Selected from H2452 Mesothelioma Cell Line

Gizem CEYLAN [1] , Sabahattin CÖMERTPAY [2]


Mezotelyoma; kalp, akciğer ve karın gibi iç organların yüzeyini kaplayan mezotelyum dokusundan gelişen bir kanser türüdür ve tümörlerinin poliKlon al kökene sahip olduğu düşünülmektedir. Kapsaisinin kanser hücreleri üzerindeki apoptotik etkileri bilinmektedir. Bu çalışmada; H2452 mezoteliyomahücre hattından seçilmiş dört (I, II, III ve IV numaralı) Klon üzerinde kapsaisinin sitotoksisitesi ve kaspaz-3 ifadesine ve aktivasyonuna etkileri tespit edilmeye çalışılmıştır. Bu amaçla; öncelikle, Klon ların IC50 değerleri MTS [3-(4,5)-dimetiltiazol-2-il)-5-(3-karboksimetoksifenil)-2-(4-sülfofenil)-2H-tetrazolyum] tuzunun formazana indirgenmesi metoduyla belirlenmiş ve kapsaisin sitotoksisitesinin bazı Klon lar arasında istatistiksel olarak anlamlı bir fark gösterdiği (p<0.01) ortaya konulmuştur. Daha sonra, seçilen Klon lara ortalama IC50 derişiminde (251.6 µM) kapsaisin uygulanmış ve hücre büyüme eğrileri çizilmiştir. Buna göre; 72 saat sonunda, Klon I’in diğer Klon lardan daha yüksek sayıda (p<0.001) canlı hücre içerdiği gözlenmiştir. Bunu takiben gerçekleştirilen qRT-PCR ve Western Blotlama ile; kapsaisin uygulanmış (KAP), etanol uygulanmış (EtOH) ve herhangi bir uygulama görmemiş, kontrol grubu hücrelerinde kaspaz-3 molekülünün sırasıyla mRNA ve protein seviyesindeki değişimler tespit edilmiştir. Sonuç olarak; kapsaisin sitotoksistesinin seçilen Klon ile anlamlı bir değişiklik gösterdiği tespit edilmiş, ve kaspaz-3 proteinin inaktif ve aktif biçimlerinin miktarı ile bağıl kaspaz-3 mRNA seviyesi her bir Klon için diğerlerinden farklı sonuçlar ortaya koymuştur. Tüm bulgular birlikte değerlendirildiğinde, mezotelyoma tümörlerini oluşturan hücre popülasyonlarının poliKlon al bir kökene sahip olabileceği fikri destekleniyor görünmektedir.

Mesothelioma is a type of cancer that develops from the mesothelium tissue that covers the surface of the internal organs such as the heart, lungs and abdomen, and its tumors are thought to have polyclonal origin. Apoptotic effects of capsaicin on cancer cells are known. In this study; we tried to determine the cytotoxicity of capsaicin and its effects on caspase-3 expression and activation on four (clone-I, -II, -III and -IV) clones selected from the H2452 mesothelioma cell line. For this purpose; firstly, capsaicin cytotoxicity, i.e. IC50 values of clones were determined by the method of reduction of MTS [3- (4,5) -dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H-tetrazolium] salt to formazan. It has been demonstrated that the type of the clones was affecting the IC50’s of capsaicin statistically significantly (p <0.01). Subsequently, capsaicin was applied to selected clones at an average concentration of IC50 (251.6 µM) and cell growth curves were plotted. Accordingly; after 72 hours of capsaicin treatment, clone-I was observed to contain a higher number of viable cells (p <0.001) than other clones. By qRT-PCR and Western blotting performed following the application, mRNA and protein levels of caspase-3 molecule were determined in capsaicin-treated (KAP), ethanol-treated (EtOH) and untreated control cells. As a result, the capsaicin cytotoxicity was found to differ significantly between the selected clones, and the amount of inactive and active forms of the caspase-3 protein and the relative caspase-3 mRNA levels showed different results for each clone. When all the findings were evaluated together, the idea that the cell populations forming mesothelioma tumors may have a polyclonal origin appears to be supported. 

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Primary Language tr
Subjects Biology
Journal Section RESEARCH ARTICLE
Authors

Orcid: 0000-0002-9627-5492
Author: Gizem CEYLAN (Primary Author)
Institution: KAHRAMANMARAŞ SÜTÇÜ İMAM ÜNİVERSİTESİ, ZİRAAT FAKÜLTESİ, TARIMSAL BİYOTEKNOLOJİ BÖLÜMÜ
Country: Turkey


Orcid: 0000-0002-2364-8171
Author: Sabahattin CÖMERTPAY
Institution: KAHRAMANMARAŞ SÜTÇÜ İMAM ÜNİVERSİTESİ, ZİRAAT FAKÜLTESİ, TARIMSAL BİYOTEKNOLOJİ BÖLÜMÜ
Country: Turkey


Supporting Institution Kahramanmaraş Sütçü İmam Üniversitesi Bilimsel Araştırma Projeleri Birimi
Project Number 2017/2-11 YLS
Thanks Bu çalışma Kahramanmaraş Sütçü İmam Üniversitesi Bilimsel Araştırma Projeleri Birimi 2017/2-11 YLS Nolu proje kapsamında gerçekleştirilmiştir.
Dates

Application Date : January 15, 2020
Acceptance Date : March 13, 2020
Publication Date : October 31, 2020

APA Ceylan, G , Cömertpay, S . (2020). Kapsaisinin H2452 Mezotelyoma Hücre Hattından Seçilen Klon larda Sitotoksisitesinin ve Kaspaz-3 Üzerine Etkilerinin Belirlenmesi . Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi , 23 (5) , 1125-1134 . DOI: 10.18016/ksutarimdoga.vi.673591