ARAŞTIRMA MAKALESİ Mersin İlinde Sağlıklı Bireylerde MikroRNA Ekspresyon Profili

Abstract

Abstract MicroRNA Expression Profiling in Healthy Subjects in Mersin Aim: MicroRNAs are a class of about 18-24 nucleotid, non-coding single-stranded small RNA molecules that regulate gene expression at the posttranscriptional level by either degrading or blocking translation of messenger RNA targets. They have roles in cardiovascular, neurodegenerative and inflammatory diseases, infections, developmental disorders, muscular disorders and cancer. The levels and types of miRNA expression in normal tissues are significantly different from in tumor tissues. The aim of this study was to investigate the profiling of circulating miRNAs expression in volunteer healthy subjects. Method: The miRNAs expression was evaluated in 292 healthy volunteers (98 women and 194 men) whose ages\'re ranging from 16 to 80. EDTA-anticoagulated peripheral blood samples were centrifuged at 4000 rpm for 15 minutes to separate plasma. cDNA's were obtained from the isolated plasma miRNAs by employing Reverse Transcription Kits and subsequently, 740 different miRNAs were analyzed with High Throughput Real-Time PCR (qRT-PCR) device (Fluidigm, Biomark, USA) by using 96.96 Dynamic Array IFCs. Results: In our study, 182 of 740 miRNAs were expressed while an expression was not observed in 558 miRNAs. Conclusion: The determination of miRNA profiles in healthy volunteers may lead to their immunotherapeutic or pharmacotherapeutic use in treatment of many diseases such as cancer in particular.

Keywords

• miRNA; healthy subjects; profiling

Mersin İlinde Sağlıklı Bireylerde MikroRNA Ekspresyon Profili

Abstract

Amaç: MikroRNA\'lar yaklaşık olarak 18-24 nükleotit uzunluğunda, tek iplikli, küçük, kodlanmayan RNA\'ların bir sınıfıdır. MikroRNA\'lar hedef mesajcı RNA\'nın yıkımını yada translasyonunun baskılanmasını gerçekleştirerek posttranskripsiyonal düzeyde gen ekspresyonunu düzenler. miRNA\'lar kardiyovasküler, nörodejeneratif ve inflamatuvar hastalıklar, infeksiyonlar, gelişimsel bozukluklar, musküler bozukluklar ve kanserde rol alırlar. Bu çalışmada sağlıklı bireylerde dolaşımda eksprese olan miRNA profilinin belirlenmesi amaçlandı. Yöntem: miRNA ekspresyonu yaşları 16 ile 80 arasında değişen, 98 kadın ve 194 erkekten oluşan 292 sağlıklı bireyde değerlendirildi. EDTA içeren tüplere alınan periferik kan örnekleri, 4000 rpm\'de 15 dakika santrifüj edilerek plazmaları ayrıldı. Plazmadan izole edilen miRNA\'lardan Reverse Transcription kitiyle cDNA\'lar elde edildi ve 96.96 Dynamic Array IFCs kullanılarak Yüksek Kapasiteli Real-Time PCR(qRT-PCR) cihazı (Fluidigm, Biomark, USA) ile 740 farklı miRNA analiz edildi. Bulgular: Çalışmamızda 740 miRNA\'dan 182\'si eksprese olurken, 558\'inde ekspresyon görülmedi. Sonuç: Sağlıklı bireylerde miRNA profilinin belirlenmesi ile başta kanser olmak üzere birçok hastalıkta immünoterapik veya farmokolojik tedavi amaçlı kullanımı gerçekleşebilir.

Keywords

• miRNA, sağlıklı birey, profil

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