Glioblastoma'da MGMT inhibitörü Diethylamine nonoate'nin epitelyal mezenkimal geçişe etkisi
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Date
2024
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Aksaray Üniversitesi Sağlık Bilimleri Enstitüsü
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info:eu-repo/semantics/openAccess
Abstract
Glioblastoma, en agresif ve en malin (grade IV) beyin tümörüdür. Glioblastoma, mevcut terapötik yaklaşımlara direnç gösterir ve bu nedenle kötü bir prognoza sahiptir. Glioblastomada kemoterapötik direncin en büyük nedeni MGMT proteinidir. MGMT inhibisyonuyla TMZ temelli alkilasyon tedavisinin başarısının artması beklenmektedir. Dietilamin NONOate (DEA/NO), nitrik oksit (NO) salınımı yapabilen bir MGMT inhibitörüdür. NO'nun, glioma hücrelerinin büyüme, invazyon, farklılaşma ve terapötik direnci etkileyebileceği belirtilmektedir. Glioblastomada terapötik dirence neden olan hücresel olaylardan biri de Epitelyal Mezenkimal Geçiş (EMT)'tir. DEA/NO'nun kanser hücrelerinde EMT yolağı üzerine etkisi bilinmemektedir. Bu tez çalışmasında DEA/NO'nun glioblastoma hücrelerinde EMT sürecinde rol alan genlerin ekspresyonuna olan etkisinin araştırılması amaçlanmıştır. Bu amaçla T98G ve U-87 MG hücre serisi kullanılmıştır. DEA/NO'nun glioma hücrelerindeki sitotoksisitesi MTT ile belirlendikten sonra hücre göçüne etkisi yara iyileşmesi testi ile belirlenmiştir. DEA/NO'nun EMT yolağında yer alan genlerin ekspresyonlarına olan etkisi qRT-PZR ile saptanmıştır. Elde edilen verilerin değerlendirilmesi amacıyla IBM SPSS Versiyon 20.0. kullanılmıştır. Sitotoksisite sonuçlarına göre U-87 MG hücre serisi T98G'ye göre daha duyarlı bulunmuştur.U87-MG hücre serisinde DEA/NO'nun hücre göçüne etkisi T98G hücre serisine göre daha yüksek bulunmuştur. DEA/NO, T98G ve U-87 MG hücre serisinde EMT genlerinin ekspresyonunu istatistiksel açsından anlamlı düzeyde azaltmıştır (p<0.05). DEA/NO glioma hücrelerinde EMT'yi indüklememektedir ve EMT genlerinin ekspresyonunu azaltmasıyla kemoterapötik ilaçlarla birlikte kullanım potansiyeline sahip olabilir. Çalışmamız, DEA/NO'nun EMT sürecini tetikleyen genlerin ekspresyonlarına olan etkisini araştıran literatürdeki ilk çalışmadır.
Glioblastoma is the most aggressive and malignant (grade IV) brain tumor. Glioblastoma shows resistance to current therapeutic approaches, resulting in a poor prognosis. The primary cause of chemotherapeutic resistance in glioblastoma is the MGMT protein. Enhancement of the success of TMZ-based alkylating therapy is expected with MGMT inhibition. Diethylenetriamine NONOate (DEA/NO) is an MGMT inhibitor capable of releasing nitric oxide (NO). It is indicated that NO may affect the growth, invasion, differentiation, and therapeutic resistance of glioma cells. One of the cellular events contributing to therapeutic resistance in glioblastoma is Epithelial-to-Mesenchymal Transition (EMT). The effect of DEA/NO on the EMT pathway in cancer cells is unknown. This thesis aims to investigate the effect of DEA/NO on the expression of genes involved in the EMT process in glioblastoma cells. For this purpose, T98G and U-87 MG cell lines were used. The cytotoxicity of DEA/NO in glioma cells was determined using MTT assay, and its effect on cell migration was determined using wound healing assay. The effect of DEA/NO on the expression of genes involved in the EMT pathway was detected by qRT-PCR. IBM SPSS Version 20.0 was used for data analysis. According to the cytotoxicity results, the U-87 MG cell line was found to be more sensitive than T98G. The effect of DEA/NO on cell migration was found to be higher in the U87-MG cell line compared to the T98G cell line. DEA/NO significantly reduced the expression of EMT genes in both T98G and U-87 MG cell lines (p<0.05). DEA/NO does not induce EMT in glioma cells and may have the potential to be used in combination with chemotherapeutic drugs by reducing the expression of EMT genes. Our study is the first in the literature to investigate the effects of DEA/NO on the expression of genes involved in the EMT process.
Glioblastoma is the most aggressive and malignant (grade IV) brain tumor. Glioblastoma shows resistance to current therapeutic approaches, resulting in a poor prognosis. The primary cause of chemotherapeutic resistance in glioblastoma is the MGMT protein. Enhancement of the success of TMZ-based alkylating therapy is expected with MGMT inhibition. Diethylenetriamine NONOate (DEA/NO) is an MGMT inhibitor capable of releasing nitric oxide (NO). It is indicated that NO may affect the growth, invasion, differentiation, and therapeutic resistance of glioma cells. One of the cellular events contributing to therapeutic resistance in glioblastoma is Epithelial-to-Mesenchymal Transition (EMT). The effect of DEA/NO on the EMT pathway in cancer cells is unknown. This thesis aims to investigate the effect of DEA/NO on the expression of genes involved in the EMT process in glioblastoma cells. For this purpose, T98G and U-87 MG cell lines were used. The cytotoxicity of DEA/NO in glioma cells was determined using MTT assay, and its effect on cell migration was determined using wound healing assay. The effect of DEA/NO on the expression of genes involved in the EMT pathway was detected by qRT-PCR. IBM SPSS Version 20.0 was used for data analysis. According to the cytotoxicity results, the U-87 MG cell line was found to be more sensitive than T98G. The effect of DEA/NO on cell migration was found to be higher in the U87-MG cell line compared to the T98G cell line. DEA/NO significantly reduced the expression of EMT genes in both T98G and U-87 MG cell lines (p<0.05). DEA/NO does not induce EMT in glioma cells and may have the potential to be used in combination with chemotherapeutic drugs by reducing the expression of EMT genes. Our study is the first in the literature to investigate the effects of DEA/NO on the expression of genes involved in the EMT process.
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Dietilamin NONOate, Epitelyal-Mezenkimal Geçiş, Hücre Kültürü, Glioblastoma, Diethylenetriamine NONOate, Epithelial-to-Mesenchymal Transition, Cell Culture, Glioblastoma
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Aksaray Üniversitesi Sağlık Bilimleri Enstitüsü
