D-vitaminin meme kanseri hücrelerinde mikro rna üzerine etkisi
Yükleniyor...
Dosyalar
Tarih
2016
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Selçuk Üniversitesi Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Vitamin D'nin aktif olan formu (1,25(OH)2D3) kalsiyum metabolizmasını düzenlemesinin yanısıra hücrenin proliferasyon, diferansiasyonunun düzenlenmesi gibi birçok fonksiyona da sahiptir. Bu fizyolojilerde oluşabilecek bozukluklar sonucunda aralarında kanser, osteoartrit, diabet gibi birçok hastalıkla da ilintili olduğu bilinmektedir. Bu çalışma, MCF7 insan meme kanseri hücresi miRNA'ları profilleri üzerine D-Vitamini uygulamasının etkilerini incelemektedir. Çalışmamızda, MCF-7 hücreleri D-vitaminin farklı saatlerde (2, 4, 6, 24 ve 48) ile tek dozda (140 uM) muamele edildi. Daha sonra total RNA, Roche yüksek saf miRNA İzolasyon Kiti (Roche Diagnostics) kullanarak izole edildi ve hücre lizatlarından cDNA sentezlendi. Gerçek zamanlı qPCR için 84 adet miRNA ekspresyonu, BioMarkTM 96,96 Dynamic Array (Fluidigm Corporation) ile saptanmıştır. İstatistiksel analiz Biogazelle en qase Plus 2,0 yazılımı kullanılarak yapıldı. Göreceli gen ekspresyonu değerlerinin tayini 2-ΔΔCt yöntemi (numunenin normalize edilmiş Ct (eşik döngüsü ) değeri- kontrolün normalize edilmiş Ct değeri) kullanılarak gerçekleştirilmiştir. İstatistiksel analizler sonucunda, kontrol grubuna kıyasla 84 miRNA'nın 22 tanesi farklı şekilde (fold regulation <2, fold regulation>2, p<0.05) eksprese edilmiştir. Özelleştirilmiş veri tabanları kullanarak hesaplanan analizlere göre (DIANA miRPath v.2.0), PI3 kinaz/AKT (hsa04151), Wnt (hsa04310), MAPK (hsa04010) ve p53 (hsa 04115) sinyal yollakları (KEGG yolak numarası) bu miRNA grubunun temel hedefleri olduğu gibi görünmektedir. Bu bulgular, MCF-7 hücrelerinin miRNA profillerine TQ'nun etkilerini vurgulamaktadır ve daha ileri çalışmalara yardımcı olabilir.
The active form of vitamin D (1,25 (OH)2D3) as well as the regulation of calcium metabolism, cell proliferation, it also has many functions, such as regulation of differentiation. These physiological disorders may occur as a result of including cancer, osteoarthritis, diabetes as are known to be associated with many diseases. This study examines the effect of Vitamin D treatment on MCF7 human breast cancer cells miRNAs. profiles. In our study, MCF-7 cells at different times of vitamin D (2, 4, 6, 24 and 48) with a single dose (140 uM) were treated. Then total RNA was isolated by using Roche High Pure miRNA Isolation Kit (Roche Diagnostics) and cDNA was synthesized from cell lysates. The expressions of 84 miRNAs were determined by The BioMarkTM 96.96 Dynamic Array (Fluidigm Corporation) for real-time qPCR. Statistical analysis was performed using the Biogazelle's qbase PLUS 2.0 software. Determinations of relative gene expression values were carried out by using the 2-∆∆Ct method (normalized threshold cycle (Ct) value of sample minus normalized Ct value of control). As a result of the statistically analysis, twenty-two of 84 miRNAs have been differentially expressed compared to control group (fold regulation <2, fold regulation>2, p<0.05). According to computational analyses using specialized databases (as DIANA miRPath v.2.0); PI3 kinase/AKT (hsa04151), Wnt (hsa04310), MAPK (hsa04010) and p53 (hsa 04115) signaling pathways (KEGG pathway number) seem to be the key targets of these miRNA group. Thesefindings highlight the effects of TQ to miRNA profiling of MCF7 cells and may be helpful for further studies.
The active form of vitamin D (1,25 (OH)2D3) as well as the regulation of calcium metabolism, cell proliferation, it also has many functions, such as regulation of differentiation. These physiological disorders may occur as a result of including cancer, osteoarthritis, diabetes as are known to be associated with many diseases. This study examines the effect of Vitamin D treatment on MCF7 human breast cancer cells miRNAs. profiles. In our study, MCF-7 cells at different times of vitamin D (2, 4, 6, 24 and 48) with a single dose (140 uM) were treated. Then total RNA was isolated by using Roche High Pure miRNA Isolation Kit (Roche Diagnostics) and cDNA was synthesized from cell lysates. The expressions of 84 miRNAs were determined by The BioMarkTM 96.96 Dynamic Array (Fluidigm Corporation) for real-time qPCR. Statistical analysis was performed using the Biogazelle's qbase PLUS 2.0 software. Determinations of relative gene expression values were carried out by using the 2-∆∆Ct method (normalized threshold cycle (Ct) value of sample minus normalized Ct value of control). As a result of the statistically analysis, twenty-two of 84 miRNAs have been differentially expressed compared to control group (fold regulation <2, fold regulation>2, p<0.05). According to computational analyses using specialized databases (as DIANA miRPath v.2.0); PI3 kinase/AKT (hsa04151), Wnt (hsa04310), MAPK (hsa04010) and p53 (hsa 04115) signaling pathways (KEGG pathway number) seem to be the key targets of these miRNA group. Thesefindings highlight the effects of TQ to miRNA profiling of MCF7 cells and may be helpful for further studies.
Açıklama
Anahtar Kelimeler
Hücre dizisi, Cell line, Hücre kültür teknikleri, Cell culture techniques, Meme neoplazmları, Breast neoplasms, Mikro RNA, Micro RNA, Neoplazmlar, Neoplasms, Polimeraz zincirleme reaksiyonu, Polymerase chain reaction, Vitamin D eksikliği, Vitamin D deficiency
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Mercan, A. (2016). D-vitaminin meme kanseri hücrelerinde mikro rna üzerine etkisi. Selçuk Üniversitesi, Yayımlanmış yüksek lisans tezi, Konya.
