KURKUMİNİN miR-145-5p VE HEDEF GENLERİ ÜZERİNDEN MEME KANSERİ HÜCRELERİNE ETKİSİ

Year 2024, Volume: 87 Issue: 3, 235 - 245, 19.07.2024

Asmaa Abuaisha , Murat Kaya , İlknur Suer , Selman Emiroğlu , Fahrünnisa Abanoz , Şükrü Palanduz , Kıvanç Çefle , Şükrü Öztürk

https://doi.org/10.26650/IUITFD.1420664

Abstract

Amaç: Kurkumin anti-kanser etkileri olan bir epigenetik regülatör olarak kabul edilmektedir. miR-145-5p, meme kanseri (MK) dahil olmak üzere bir çok kanserde ekspresyon düzeyi düşük olan bir tümör baskılayıcı mikroRNA'dır (miRNA). Bu çalışmada, kurkuminin miR-145-5p’yi ve olası hedef genlerini regüle ederek MCF-7 insan MK hücre hattının proliferasyonunu ve göçünü inhibe edip etmediğinin araştırılması amaçlanmıştır.
Gereç ve Yöntem: MCF-7 hücreleri kurkumin ve onun solvent kontrolü ile muamele edildi. Ayrıca hücreler miR-145-5p mimiği ve negatif kontrol miRNA mimiği ile transfekte edildi. Hücre canlılığı değerlendirildi ve hücre göçü scratch yara (wound) testi kullanılarak değerlendirildi. miR-145-5p’nin potansiyel hedefleri, MK doku örneklerinde aşırı eksprese edilen genlerle miRNet ve miRTarBase v8 veri-tabanları ile örtüşen genlerin Kanser Genom Atlası (TCGA) datasetlerinde araştırılması yoluyla belirlendi. miR-145-5p ve seçilen genlerin ekspresyon düzeyleri, kantitatif gerçek-zamanlı polimeraz zincir reaksiyonu (qRT-PCR) ile tespit edildi. Kantifikasyon analizi için 2-ΔΔCt yöntemi kullanıldı. p-değeri <0,05 istatistiksel olarak anlamlı kabul edildi.
Bulgular: Hem kurkumin ile muamele edilmiş hem de miR-145-5p mimiği ile transfekte edilmiş MCF-7 hücrelerinde, proliferasyonun ve göçün, kontrollere kıyasla azaldığı gözlenmiştir. Ayrıca, kurkumin ile muamele edilen hücrelerde miR-145-5p'nin ekspresyonunun anlamlı düzeyde arttığı görülmüştür. miR-145-5p’nin olası hedef genlerinin, MCM2, MMP1, MMP9 ve EEF1A2, kurkumin ile muamele edilmiş MCF-7 hücrelerinde ekspresyon düzeyinin azaldığı saptanmıştır. Bunun yanı sıra, miR-145-5p mimiği ile transfekte edilmiş hücrelerde, MCM2, MMP1 ve MMP9 genlerinin ekspresyon düzeyinin düşük olduğu belirlenmiştir.
Sonuç: Kurkuminin, miR-145-5p ve olası hedef genleri üzerinden etki ederek MCF-7 hücrelerinin hem proliferasyonunu hem de göçünü inhibe ettiği gösterilmiştir.

Keywords

Meme kanseri, MCF-7 hücre hattı, kurkumin, miR-145-5p, hedef genler

EFFECT of CURCUMIN on BREAST CANCER CELLS THROUGH miR-145-5p AND ITS TARGET GENES

Abstract

Objective: Curcumin is considered an epigenetic regulator with anticancer effects. The micro-RNA (miRNA) miR-145-5p is a tumor suppressor that shows low expression levels in various cancers, including breast cancer (BC). This study aims to investigate whether curcumin inhibits MCF-7 human BC cell line proliferation and migration by regulating miR-145-5p and its possible target genes.
Material and Method: MCF-7 cells were treated with curcumin and its solvent control. Additionally, cells were transfected with an miR-145-5p mimic and a non-targeting miRNA mimic. Cell viability was then evaluated, and the scratch wound assay was used to assess cell migration. The study predicts the miR-145- 5p putative target genes by searching for overlapping genes in the miRNet and miRTarBase v8 databases via the overexpressed genes in the BC tissue samples in the Cancer Genome Atlas (TCGA) datasets. Expression levels of miR-145-5p and the selected genes were detected using the quantitative real-time polymerase chain reaction (qRT-PCR). The 2-ΔΔCt method was used for the quantification analysis, with p<0.05 being considered statistically significant.
Result: Curcumin treatment and overexpression of miR-145- 5p via the transfection of an miR-145-5p mimic significantly decreased the proliferation and migration of MCF-7 cells. Moreover, curcumin treatment significantly increased the expression of miR-145-5p. The possible target genes of miR-145- 5p (i.e., MCM2, MMP1, MMP9, EEF1A2) were downregulated in curcumin-treated MCF-7 cells. Additionally, miR-145-5p mimictransfected cells showed low expression levels of the MCM2, MMP1, and MMP9 genes.
Conclusion: Curcumin inhibits the proliferation and migration of MCF-7 cells by acting on miR-145-5p and its possible target genes.

Keywords

Breast cancer, MCF-7 cell line, curcumin, miR-145- 5p, target genes

Ethical Statement

No ethics committee approval was need for this study, since the study was conducted on human breast cancer cell line and public available datasets.

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