OVER KANSERİ HÜCRELERİNDE PRİMA-1 Met TEDAVİSİNE YANIT OLARAK DEĞİŞEN miRNA EKSPRESYON ANALİZİ

Abstract

Tümör hücrelerinde p53 fonksiyonunun restorasyonu, over kanseri tedavisinde çekici bir strateji olacağı düşünülmektedir, çünkü p53 mutasyonlarının over kanserlerinde görülme sıklığı  %50-60 arasındadır. Küçük molekül Prima-1^Met'in, p53'ün tümör baskılama fonksiyonunu geri kazandığı ve insan tümör hücrelerinde hücre büyümesini inhibe ettiği ve apoptozu indüklediği gösterilmiştir. MikroRNA'lar hem transkripsiyonel hem de translasyonel seviyelerde gen ekspresyonunu düzenler ve hücre proliferasyonu, farklılaşma ve hematopoez gibi çok çeşitli fizyolojik ve biyolojik süreçlerde etki yapar. Epitelyal over kanserinde yapılan çok sayıdaki miRNA profillemesi çalışmalarında, kemoterapi direnci ve hastalık progresyonu ile ilişkili miRNA'lar tanımlanmıştır, fakat, Prima-1^Met'e yanıt olarak miRNA'ların tutulumu hakkında çok az şey bilinmektedir. Bu çalışmada, apoptotik etkisi olduğu bilinen Prima-1^Met ile muamele edilmiş over kanseri hücre hatlarında, bu ilaca yanıt olarak ekspresyonu değişen miRNA’ların belirlenmesini hedeflendi ve bunun için ilaç verilen hücre hatlarında hem kanser hem de apoptosis yolaklarını hedefleyen miRNA’ların ekspresyonları miScript PCR array ile belirlenip analiz edilmiştir. Analiz sonucunda, her iki hücre hattında da hem over kanseri hem de apoptosisle ilişkili olarak Prima-1^Metuygulamasıyla ekspresyonu artan miRNA’lar; miRNA-1, miRNA-134, miRNA-141, miRNA-143, miRNA-145, miRNA-204, miRNA-205, miRNA-214, miRNA-29a ve miRNA-29c olarak belirlenmiştir. Ekspresyonu azalan miRNA’lar ise miRNA-21, miRNA-221 ve miRNA-222 olarak tespit edilmiştir. Bu çalışma Prima-1^Met indüklü apoptosisin moleküler mekanizmasının aydınlatılması için bir temel oluşturmaktadır. 

Keywords

• Prima-1Met,miRNA,PCR-array,over kanseri,ekspresyon

ANALYSIS OF DIFFERENTIAL miRNA EXPRESSION IN RESPONSE TO PRIMA-1 Met THERAPY IN OVARIAN CANCER CELLS

Abstract

Restoration of p53 function in tumor cells will be an attractive strategy for ovarian cancer therapy since p53 mutations are found in more than 50 % of ovarian cancers. The small molecule Prima-1^Met has been shown to restore tumor suppression function of p53, inhibit cell growth and induce apoptosis in human tumor cells. MicroRNAs (miRNAs) regulate gene expression at both transcriptional and translational levels as well as acting in a wide variety of physiological and biological processes. Numerous miRNAs associated with chemotherapy resistance and disease progression have been described in a large number of miRNA profiling studies in epithelial ovarian cancer, but little is known about the involvement of miRNAs in response to Prima-1^Met. We aimed to determine the expression of miRNAs in response to Prima-1^Met treatment, which is known to have apoptotic effects in ovarian cancer cell lines. The expressions of miRNAs targeting both cancer and apoptosis pathways in drug-delivered cell lines were determined and analyzed by miScript PCR array. As a result of the assay, the expression of miRNA-1, miRNA-134, miRNA-141, miRNA-143, miRNA-145, miRNA-204, miRNA-205, miRNA-214, miRNA-29a and miRNA-29c increased in response to Prima-1^Met in both cell lines, while the expression of miRNA-21, miRNA-221 and miRNA-222 was reduced. In conclusion, this study provides a basis for elucidating the molecular mechanism of Prima-1^Met-induced apoptosis.

Keywords

• Prima-1Met,miRNA,PCR-array,ovarian cancer,expression

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