Yıl: 2021 Cilt: 9 Sayı: 2 Sayfa Aralığı: 166 - 173 Metin Dili: İngilizce DOI: 10.4274/nkmj.galenos.2021.770512 İndeks Tarihi: 12-10-2021

The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line

Öz:
Aim: It was aimed to investigate the effect of miR-34a-5p and miR-145-5p on breast cancer cell line MDA-MB-231 and to determine the expressionof target genes of these microRNAs (miRNAs).Materials and Methods: Firstly, literature search and in silico analysis were performed to detect possible target genes of miR-34a-5p and miR-145-5p, which are known to be tumor suppressors. Mimic miR-34a-5p and miR-145-5p were transfected to the breast cancer cell line MDA-MB-231.Deregulated genes were investigated by the quantitative real-time polymerase chain reaction compared to control cells. Also, the effect of thesemiRNAs on proliferation was determined using the Water Soluble Tetrazolium Salt-8 method. Finally, the expressions of epithelial mesenchymaltransition (EMT) markers, which are known to be important in the metastatic process, are examined.Results: The proliferation of the miR-34a-5p or miR-145-5p transfected cells decreased compared to the control groups. The expression of E2Ftranscription factor 1 (E2F1) (p=0.009), mitogen activated protein kinase 1 (MEK1) (p=0.001) and cyclin dependent kinase 4 (CDK4) (p=0.005)genes, which were among the genes targeted by miR-34a-5p, were significantly reduced. EMT markers were significantly changed in miR-34a-5ptransfected cells (E-Cad increase p=0.01; Vimentin decrease p=0.008). Kruppel-like factor 4 (KLF4) (p=0.007) targeted miR-145-5p were significantlyreduced and EMT markers were significantly changed in miR-145-5p transfected cells (E-Cad increase p=0.0005; Vimentin decrease p=0.006).Conclusion: miR-34a-5p and miR-145-5p may have an impact on the breast cancer cell line MDA-MB-231 proliferation and EMT mechanism. Atthe same time, according to our study results, it was revealed that E2F1, MEK1 and CDK4 genes, whose expression level decreased after transfectionof mimic miR-34a-5p, could be targeted by miR-34a-5p in breast cancer, and that the expression level of KLF4, which decreased as a result of mimicmiR-145-5p transfection, could be the target.
Anahtar Kelime:

miR-34a-5p ve miR-145-5p’nin MDA-MB-231 Hücre Hattındaki Ektopik Ekspresyonunun Hücre Proliferasyonu ve Hedef Gen Ekpresyonu Üzerindeki Etkisi

Öz:
Amaç: Bu çalışmada miR-34a-5p ile miR-145-5p’nin meme kanseri hücre hattı MDA-MB-231 üzerindeki fonksiyonel etkilerinin araştırılması ve bu mikroRNA’ların (miRNA) olası hedef genlerinin miRNA mimik transfeksiyonu yapılan hücrelerde ifade seviyelerinin araştırılması hedeflenmiştir. Gereç ve Yöntem: Tümör baskılayıcı özellikte oldukları bilinen miR-34a-5p ve miR-145-5p’nin meme kanserindeki olası hedef genlerinin tespiti için öncelikle literatür taraması ve in silico analiz çalışması yapılmıştır. Tespit edilen genlerin ifadelerindeki değişimler in vitro ortamda meme kanseri hücre hattı MDA-MB-231’de belirlenen miRNA’ların transfekte edilmesiyle kontrol hücrelerle karşılaştırmalı olarak kantitatif gerçek zamanlı polimeraz zincir reaksiyonu ile araştırılması hedeflenmiştir. Ayrıca bu miRNA’ların proliferasyona etkisi Water Soluble Tetrazolium Salt-8 yöntemi kullanılarak belirlenmiştir. Son olarak metastatik süreçte önemli olduğu bilinen epitelyal mezenkimal geçiş (EMT) belirteçlerinin ekspresyonuna bakılarak metastaza olan katkısının araştırılması hedeflenmiştir. Bulgular: miR-34a-5p transfeksiyonu yapılan gruptaki hücre proliferasyonunun kontrol grubu hücrelere göre azaldığı, EMT ile ilişkili genlerin ifadesinin anlamlı şekilde değiştiği (E-Cad artış p=0,01; Vimentin azalma p=0,008), miR-34a-5p’nin hedeflediği genlerden olan E2F transkripsiyon faktör 1 (E2F1) (p=0,009), mitojen aktive protein kinaz 1 (MEK1) (p=0,001) ve siklin bağımlı kinaz 4 (CDK4) (p=0,005) genlerinin ifadesinin belirgin şekilde düştüğü tespit edilmiştir. miR-145-5p transfekte edilen hücrelerde ise benzer şekilde hücre proliferasyonunda azalma ve EMT ile ilişkili genlerde anlamlı değişiklikler gözlenmiş (E-Cad artış p=0,0005; Vimentin azalma p=0,006), miR-145-5p’nin hedeflediği önemli proto onkogenlerden birisi olan Kruppel-benzeri faktör 4 (KLF4) (p=0,007) geninin ifadesinin önemli derecede azaldığı tespit edilmiştir. Sonuç: Çalışma sonuçlarımız, miR-34a-5p ve miR-145-5p’nin meme kanseri hücre hattı MDA-MB-231 hücre proliferasyonu ve EMT mekanizması üzerinde etkili olabileceğini göstermiştir. Aynı zamanda çalışma sonuçlarımıza göre mimik miR-34a-5p transfeksiyon yapılması sonrasında ekspresyon seviyesi azalan E2F1, MEK1 ve CDK4 genlerinin meme kanserinde miR-34a-5p tarafından hedeflenebileceği, yine mimik miR-145-5p transfeksiyonu neticesinde ifade seviyesi azalan KLF4 geninin miR-145-5p’nin hedefi olabileceği ortaya çıkmıştır.
Anahtar Kelime:

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık
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APA SUER İ, KAYA M, Özgür E (2021). The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line. , 166 - 173. 10.4274/nkmj.galenos.2021.770512
Chicago SUER İlknur,KAYA Murat,Özgür Emre The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line. (2021): 166 - 173. 10.4274/nkmj.galenos.2021.770512
MLA SUER İlknur,KAYA Murat,Özgür Emre The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line. , 2021, ss.166 - 173. 10.4274/nkmj.galenos.2021.770512
AMA SUER İ,KAYA M,Özgür E The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line. . 2021; 166 - 173. 10.4274/nkmj.galenos.2021.770512
Vancouver SUER İ,KAYA M,Özgür E The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line. . 2021; 166 - 173. 10.4274/nkmj.galenos.2021.770512
IEEE SUER İ,KAYA M,Özgür E "The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line." , ss.166 - 173, 2021. 10.4274/nkmj.galenos.2021.770512
ISNAD SUER, İlknur vd. "The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line". (2021), 166-173. https://doi.org/10.4274/nkmj.galenos.2021.770512
APA SUER İ, KAYA M, Özgür E (2021). The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line. Namık Kemal Tıp Dergisi, 9(2), 166 - 173. 10.4274/nkmj.galenos.2021.770512
Chicago SUER İlknur,KAYA Murat,Özgür Emre The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line. Namık Kemal Tıp Dergisi 9, no.2 (2021): 166 - 173. 10.4274/nkmj.galenos.2021.770512
MLA SUER İlknur,KAYA Murat,Özgür Emre The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line. Namık Kemal Tıp Dergisi, vol.9, no.2, 2021, ss.166 - 173. 10.4274/nkmj.galenos.2021.770512
AMA SUER İ,KAYA M,Özgür E The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line. Namık Kemal Tıp Dergisi. 2021; 9(2): 166 - 173. 10.4274/nkmj.galenos.2021.770512
Vancouver SUER İ,KAYA M,Özgür E The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line. Namık Kemal Tıp Dergisi. 2021; 9(2): 166 - 173. 10.4274/nkmj.galenos.2021.770512
IEEE SUER İ,KAYA M,Özgür E "The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line." Namık Kemal Tıp Dergisi, 9, ss.166 - 173, 2021. 10.4274/nkmj.galenos.2021.770512
ISNAD SUER, İlknur vd. "The Effect of miR-34a-5p and miR-145-5p Ectopic Expression on Cell Proliferation and Target Gene Expression in the MDA-MB-231 Cell Line". Namık Kemal Tıp Dergisi 9/2 (2021), 166-173. https://doi.org/10.4274/nkmj.galenos.2021.770512