ZN(II) kompleksinin sitotoksik ve apoptotik etkilerinin kanser hücrelerinde araştırılması
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Date
2019-09-24
Authors
Alioğlu, İmren
Journal Title
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Publisher
Bursa Uludağ Üniversitesi
Abstract
Kanser, hücrelerin kontrolsüz çoğalması ile oluşan bir hastalıktır. Hücrelerin çeşitli risk faktörleri sebebiyle mutasyona uğrayıp sınırsız çoğalması ile başlayan bu süreç sonunda kanserli hücreler oluşmakta, dokulara ve organizmaya zarar vermektedir. Son yıllarda yapılan çalışmalar, kanser tedavisinde metal bazlı bileşiklerin umut verici olduğunu göstermiştir. Özellikle platin (Pt), gümüş (Ag), altın (Au) ve çinko (Zn) gibi metal kompleksleri birçok antikanser araştırmalarda kullanılmış ve başarılı sonuçlar alınmıştır. Dolayısıyla bu tez çalışmasında, bir Zn(II) kompleksinin insan meme kanseri hücrelerinde antikanser etkileri ve bu etkilerden sorumlu mekanizmalar araştırılmıştır. Zn(II) kompleksinin insan meme kanseri hücrelerindeki anti proliferatif potansiyeli SRB canlılık testi ile belirlenmiş, hücre canlılıkları ATP canlılık testi ile doğrulanmıştır. Ayrıca kompleksin olası sitotoksik etkisi sağlıklı insan meme hücrelerde de değerlendirilmiştir. Sitotoksik etkilerinden sorumlu olan hücre ölüm mekanizması ve apoptoz varlığını göstermek için Hoechst 33342/Propidyum İyodür ikili boyama yöntemi ve kırılmış sitokeratin-18 (M30 antijen seviyesi) yöntemi kullanılmıştır. Hücre ölümü ile ilişkili proteinlerin ekspresyon seviyeleri (B-Aktin, PARP/kırılmış PARP, COX4, DR4, Prokaspas-8/kırılmış Kaspas-8, BAX, RIP) immunoblotlama yöntemi ile gösterilmiştir. Apoptozisle ilişkili gen ekspresyonları ise Polimeraz Zincir Reaksiyonu kullanılarak belirlenmiştir. Zn(II) kompleksinin hücre migrasyonu üzerine etkisini belirlemek amacıyla yara iyleşmesi testi uygulanmıştır. Ayrıca Zn(II) kompleksinin kanser kök hücrelerindeki sitotoksik etkisi kök hücre populasyonunca zengin olan mamosferler üzerinde araştırılmıştır. Elde edilen sonuçlarda, Zn(II) kompleksinin insan meme kanseri hücrelerinde doza ve zamana bağlı olarak sitotoksik etkiye neden olduğu sağlıklı hücrelerde ise toksik olmadığı gözlendi. Zn(II) kompleksinin kanser hücrelerinde apoptozisi indüklediği hem protein hem de gen düzeyinde belirlendi. Zn(II) kompleksinin kanser hücrelerinin migrasyon yeteneklerini önemli derecede engellediği gözleniği, ayrıca kanser kök hücrelerinde de sitotoksik aktiviteye sahip olduğu belirlenmiştir. Tüm sonuçlar değerlendirildiğinde, Zn(II) kompleksinin kanser tedavisinde umut verici bir ajan olarak kullanılabileceği öngörüsüyle ileri analizlerin yapılması gerektiği sonucuna varılmıştır.
Cancer is a disease caused by uncontrolled proliferation of cells. This process begins with the mutation of cells due to various risk factors and unlimited proliferation. Recent studies have shown that metal-based compounds are promising in the treatment of cancer. Especially metal complexes such as platinum (Pt), silver (Ag), gold (Au) and zinc (Zn) have been used in many anticancer studies and successful results have been obtained. Therefore, the anticancer effects of a Zn(II) complex in human breast cancer cells and the mechanisms responsible for these effects were investigated in this thesis. The anti-proliferative potential of Zn(II) complex in human breast cancer cells was determined by SRB viability test. Cell viability was confirmed by ATP viability test. The possible cytotoxic effect of the complex was also evaluated in healthy human breast cells. Hoechst 33342 / Propidium Iodide dual staining method and broken cytokeratin-18 (M30 antigen level) method were used to demonstrate the presence of cell death mechanism and apoptosis responsible for cytotoxic effects. Expression levels of cell death-related proteins (B-Actin, PARP/cleaved PARP, COX4, DR4, Prokaspas-8 / cleaved Kaspas-8, BAX, RIP) were demonstrated by immunoblotting. Apoptosis-related gene expression was determined using Polymerase Chain Reaction. Wound healing test was performed to determine the effect of Zn(II) complex on cell migration. In addition, the cytotoxic effect of Zn(II) complex on cancer stem cells was investigated on mamospheres rich in stem cell population. The results showed that Zn(II) complex caused cytotoxic effect in human breast cancer cells depending on dose and time and was not toxic in healthy cells. Zn(II) complex induces apoptosis in cancer cells was determined at both protein and gene levels. Zn(II) complex significantly inhibited the migration ability of cancer cells. Zn(II) complex also had cytotoxic activity in cancer stem cells. When all the results were evaluated, it was concluded that further analysis should be performed with the assumption that Zn(II) complex can be used as a promising agent in the treatment of cancer.
Cancer is a disease caused by uncontrolled proliferation of cells. This process begins with the mutation of cells due to various risk factors and unlimited proliferation. Recent studies have shown that metal-based compounds are promising in the treatment of cancer. Especially metal complexes such as platinum (Pt), silver (Ag), gold (Au) and zinc (Zn) have been used in many anticancer studies and successful results have been obtained. Therefore, the anticancer effects of a Zn(II) complex in human breast cancer cells and the mechanisms responsible for these effects were investigated in this thesis. The anti-proliferative potential of Zn(II) complex in human breast cancer cells was determined by SRB viability test. Cell viability was confirmed by ATP viability test. The possible cytotoxic effect of the complex was also evaluated in healthy human breast cells. Hoechst 33342 / Propidium Iodide dual staining method and broken cytokeratin-18 (M30 antigen level) method were used to demonstrate the presence of cell death mechanism and apoptosis responsible for cytotoxic effects. Expression levels of cell death-related proteins (B-Actin, PARP/cleaved PARP, COX4, DR4, Prokaspas-8 / cleaved Kaspas-8, BAX, RIP) were demonstrated by immunoblotting. Apoptosis-related gene expression was determined using Polymerase Chain Reaction. Wound healing test was performed to determine the effect of Zn(II) complex on cell migration. In addition, the cytotoxic effect of Zn(II) complex on cancer stem cells was investigated on mamospheres rich in stem cell population. The results showed that Zn(II) complex caused cytotoxic effect in human breast cancer cells depending on dose and time and was not toxic in healthy cells. Zn(II) complex induces apoptosis in cancer cells was determined at both protein and gene levels. Zn(II) complex significantly inhibited the migration ability of cancer cells. Zn(II) complex also had cytotoxic activity in cancer stem cells. When all the results were evaluated, it was concluded that further analysis should be performed with the assumption that Zn(II) complex can be used as a promising agent in the treatment of cancer.
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Keywords
Meme kanseri, Zn(II) complex, Zn(II) kompleksi, Apoptozis, Breast cancer, Apoptosis, Cytotoxic, Sitotoksite
Citation
Alioğlu, İ. (2019). ZN(II) kompleksinin sitotoksik ve apoptotik etkilerinin kanser hücrelerinde araştırılması. Yayınlanmamış yüksek lisans tezi. Bursa Uludağ Üniversitesi Fen Bilimleri Enstitüsü.