Multiple myeloma kanser hücrelerinde bortezomibe karşı oluşan çoklu ilaç direnç mekanizmasının araştırılması
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Dosyalar
Tarih
2019
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Yayıncı
Aksaray Üniversitesi Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Multiple Myeloma (MM), kemik iliğinde malign plazma hücrelerinin birikmesiyle karakterize olan hematolojik bir kanserdir. Bortezomib myeloma tedavisinde kullanılan en etkin kemoterapötik ilaçtır. Fakat tedavi sürecinde bortezomibe karşı gelişen ilaç dirençliliği, kemoterapide başarıya ulaşmayı engellemektedir. Kanser hücrelerinde çoklu ilaç dirençlilik genlerinin (MDR) ifadesinin artması ilaç dirençliliğine sebep olan en önemli faktörlerden birisidir. Bu nedenle tez çalışmasında, MM hücrelerinde ilaç dirençliliğiyle ilişkili olan P-gp, MRP-1, MRP-2, MRP-3, MRP-6, MRP-7 ilaç taşıyıcı protein genlerinin ve GSTP1'in ifade düzeyleri araştırılmıştır. Bu çalışmada, multiple myeloma KMS20 (bortezomibe dirençli) ve KMS28 (bortezomibe hassas) hücre hatlarına MTT testi uygulanarak bortezomibin IC50 değerleri belirlenmiştir. Her iki hücre hattından RNA izolasyonları yapılarak cDNA'lar elde edilmiştir. Çalışılan genlerin ifade düzeyleri qRT-PCR ile analiz edilmiştir. Gen ifade analizleri sonucunda MM'de bortezomib dirençliliğinden esas sorumlu taşıyıcının P-gp (MDR-1) olduğu bulunmuştur. Bortezomibin büyük olasılıkla GSTP1 ile glutatyonlanarak MRP taşıyıcılarıyla hücreden atıldığı düşünülmektedir. MRP-7'nin bortezomib dirençliliğinden sorumlu olduğu ilk kez bu çalışmayla açığa çıkmıştır. Ayrıca uzun süre yüksek bortezomib dozuna maruz kalmayla MRP-1 ifadesinin ortaya çıktığı ve bu durumdaki MRP-2'nin bortezomib dirençliliği gelişiminde kısmen etkili olduğu da tespit edilmiştir. MRP-6 ifadesinin bortezomib dirençliliğiyle ilişkisinin olmadığı ilk kez bu çalışmayla gösterilmiştir. MRP-3 ifadesi kullanılan her iki hücre hattında da tespit edilmemiştir. Bu sonuçlar doğrultusunda ifadesi yüksek olan genler uygun siRNA'lar veya baskılayıcı moleküller kullanılarak engellenebilir. Multiple myeloma için bu çalışmayla birlikte diğer tüm dirençlilik mekanizmalarının aydınlatılmasıyla tedavi şekillerinin kişiye özel olarak geliştirilmesi de mümkün olabilecektir.
Multiple Myeloma (MM) is a hematological cancer characterized by accumulation of malignant plasma cells and bortezomib is the most effective chemotherapeutic used in treatment. However, drug resistance prevents success of chemotherapy in treatment process. One of the factors causing resistance is overexpression of multiple drug resistance genes (MDR). Therefore, in this study expression levels of MDR-1 (Pgp), MRP-1, MRP-2, MRP-3, MRP-6, MRP-7 and GSTP-1 genes in MM cell lines were investigated. IC50 values of bortezomib were determined by MTT assay in KMS20 (bortezomib resistant) and KMS28 (bortezomib sensitive) multiple myeloma cell lines. RNA was isolated from both cell lines and cDNAs were obtained. Expression levels of investigating genes were analyzed by qRT-PCR. As a result of gene expression analysis, it was found that P-gp (MDR-1) was the main responsible transporter for bortezomib resistance in MM. Bortezomib is strong probably thought to be excreted from the cell with MRP transporters by glutathione with GSTP1. This was the first time that MRP-7 expression was responsible for bortezomib resistance. In addition, MRP-1 expression appears with prolonged exposure to high bortezomib doses and in the same condition MRP-2 is partially effective in the development of bortezomib resistance. This is the first report that MRP-6 expression was not associated with bortezomib resistance. MRP-3 expression was not detected in both cell lines. According to the results of this study, genes with high expression can be prevented by using appropriate siRNAs or suppressive molecules. Through the results of this study and solving all other resistance mechanisms for multiple myeloma are be possible to develop personalized treatment forms.
Multiple Myeloma (MM) is a hematological cancer characterized by accumulation of malignant plasma cells and bortezomib is the most effective chemotherapeutic used in treatment. However, drug resistance prevents success of chemotherapy in treatment process. One of the factors causing resistance is overexpression of multiple drug resistance genes (MDR). Therefore, in this study expression levels of MDR-1 (Pgp), MRP-1, MRP-2, MRP-3, MRP-6, MRP-7 and GSTP-1 genes in MM cell lines were investigated. IC50 values of bortezomib were determined by MTT assay in KMS20 (bortezomib resistant) and KMS28 (bortezomib sensitive) multiple myeloma cell lines. RNA was isolated from both cell lines and cDNAs were obtained. Expression levels of investigating genes were analyzed by qRT-PCR. As a result of gene expression analysis, it was found that P-gp (MDR-1) was the main responsible transporter for bortezomib resistance in MM. Bortezomib is strong probably thought to be excreted from the cell with MRP transporters by glutathione with GSTP1. This was the first time that MRP-7 expression was responsible for bortezomib resistance. In addition, MRP-1 expression appears with prolonged exposure to high bortezomib doses and in the same condition MRP-2 is partially effective in the development of bortezomib resistance. This is the first report that MRP-6 expression was not associated with bortezomib resistance. MRP-3 expression was not detected in both cell lines. According to the results of this study, genes with high expression can be prevented by using appropriate siRNAs or suppressive molecules. Through the results of this study and solving all other resistance mechanisms for multiple myeloma are be possible to develop personalized treatment forms.
Açıklama
Anahtar Kelimeler
Multiple Myeloma, Bortezomib, İlaç Dirençliliği, MDR
