AIFM3 ARACILI APOPTOZ SİNYALİNİN KANSER HÜCRELERİNDE MİR-96-5P İLE BASKILANMASI

Amaç: Kanser dünya genelinde yaygın olarak görülen önemli bir halk sağlığı problemidir. Mikro RNA’lar (miRNA), kanser gibi karmaşık hastalıkların moleküler mekanizmasında önemli fonksiyonları olan küçük kodlanmayan RNA transkriptleridir. Bu çalışmada, miR-96-5p’nin meme, akciğer ve osteosarkom kanser hücrelerinde AIFM3 (apoptosis inducing factor mitochondria associated 3) geni ve apoptoz üzerine olan etkilerinin araştırılması amaçlanmıştır. Materyal ve Metot: Osteosarkom (U2-OS), meme (MCF7) ve akciğer (A549) kanseri hücreleri normal kültür koşullarında çoğaltılıp ardından miR-96-5p mimik transfeksiyonu yapılmıştır. Mimik transfekte edilen hücreler toplanarak miRNA ve total RNA izolasyonları yapılmıştır. Real-Time PCR ile gen ifadesi analizleri gerçekleştirilmiştir. Bulgular: Kanser hücrelerinde miR-96-5p mimik transfeksiyonu sonrasında AIFM3 geninin ifade düzeyinin meme (p=0,015) ve akciğer (p=0,024) kanseri hücrelerinde ciddi derecede azaldığı gösterilmiştir. Osteosarkom hücrelerinde ise miR-96-5p mimik transfeksiyonu sonrasında AIFM3 geninin ifade düzeyinde azalma olduğu gösterilmiştir, ancak bu değişim istatistiksel olarak anlamlı değildir. Ayrıca, bu hücrelerde mimik transfeksiyonu sonrasında apoptoz ile ilişkili genlerin ifade düzeylerinde önemli değişikliklerin olduğu saptanmıştır. MiR-96-5p mimik transfeksiyonu sonrasında, MCF7 hücrelerinde BCLxL (p=0,043) ve BCL2 (p=0,0147) genlerinin, A549 akciğer kanseri hücrelerinde BCL2 (p=0,026) geninin ifade düzeyinin arttığı bulunmuştur. Sonuç: Sonuç olarak, kanser hücrelerinde miR-96-5p’nin ifade düzeyinin arttırılması ile AIFM3 geninin ifade düzeyinin azaldığı ve apoptoz ilişkili sinyal yolağının baskılandığı bulunmuştur. MiR-96-5p’nin apoptoz üzerine etkilerin net olarak anlaşılması için daha kapsamlı çalışmalara ihtiyaç vardır.

Suppression of AIFM3-Mediated Apoptosis Signal by miR-96-5p in Cancer Cells

Aim: Cancer is an important public health problem that is common throughout the world. Micro RNAs (miRNAs) are small non-coding RNA transcripts that have important functions in the molecular mechanism of complex diseases such as cancer. In this study, it was aimed to reveal the effects of miR-96-5p on AIFM3 (apoptosis inducing factor mitochondria associated 3) gene and apoptosis in breast, lung and osteosarcoma cancer cells. Materials and Methods: Osteosarcoma (U2-OS), breast (MCF7) and lung (A549) cancer cells were grown under normal culture conditions and subsequently subjected to miR-96-5p mimic transfection. Mimic transfected cells were harvested and miRNA and total RNA isolations were performed. Gene expression analyzes were performed by Real-Time PCR. Results: Expression of AIFM3 gene was shown to be significantly reduced in breast (p=0.015) and lung (p=0.024) cancer cells after miR-96-5p mimic transfections. In osteosarcoma cells, the expression level of the AIFM3 gene was shown to be decreased after miR-96-5p mimic transfections, but this change was not statistically significant. In addition, significant changes were found in the expression levels of apoptosis-related genes after mimic transfection in cancer cells. After miR-96-5p mimic transfection, the expression level of BCLxL (p = 0.043) and BCL2 (p = 0.0147) genes in MCF7 cells and BCL2 (p = 0.026) gene in A549 lung cancer cells were found to be increased. Conclusion: Consequently, it was found that by increasing the expression level of miR-96-5p in cancer cells, the expression level of AIFM3 gene decreased and apoptosis-associated signaling was suppressed. Further more comprehensive studies are needed to clearly understand the effects of miR-96-5p on apoptosis.

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