MiR-96-5p suppress apoptosis-related signalling by regulating AIFM3 in cancer cells

Abstract

Objective: Cancer is an important public health problem that is common throughout the world. MicroRNAs (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 the effects of miR-96-5p on apoptosis inducing factor mitochondria associated 3 (AIFM3) gene and apoptosis in breast, lung and osteosarcoma cancer cells. Materials and Methods: 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 and lung cancer cells after miR-96-5p mimic transfection. In addition, significant changes in expression levels of apoptosis-related genes were found after mimic transfection in these cells. 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.

Keywords

• AIFM3,apoptosis,cancer

Kanser hücrelerinde miR-96-5p’nin AIFM3’ü düzenleyerek apoptoz ilişkili sinyali baskılaması

Öz

Amaç: Kanser dünya genelinde yaygın olarak görülen önemli bir halk sağlığı problemidir. MikroRNA’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 mitokondri ilişkili apoptoz uyarıcı faktör 3 (AIFM3) geni ve apoptoz üzerine olan etkilerinin araştırılması amaçlanmıştır. Materyal ve Metot: Kanser hücreleri normal kültür koşullarında çoğaltılıp ardından miR-96-5p mimik transfeksiyonu yapılmıştır. Mimik transfekte edilen hücreler toplanarak miRNA ve toplam 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 ve akciğer kanseri hücrelerinde ciddi derecede azaldığı gösterilmiştir. Ayrıca, bu hücrelerde mimik transfeksiyonu sonrasında apoptoz ile ilişkili genlerin ifade düzeylerinde önemli değişikliklerin olduğu saptanmıştır. 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.

Anahtar Kelimeler

• AIFM3,apoptoz,kanser

References

1. 1. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2019. CA: a cancer journal for clinicians. 2019;69(1):7-34.
2. 2. Dalmay T, Edwards D. MicroRNAs and the hallmarks of cancer. Oncogene. 2006;25(4):6170-5.
3. 3. Yu J-J, Wu Y-X, Zhao F-J, Xia S-J. miR-96 promotes cell proliferation and clonogenicity by down-regulating of FOXO1 in prostate cancer cells. Med. Oncol. 2014;31(4):910.
4. 4. Bao Y, Wang Y, Liu Y, Wang S, Wu B. MiR-96 expression in prostate cancer and its effect on the target gene regulation. Eur Rev Med Pharmacol Sci. 2017;21(20):4548-6.
5. 5. Iwai N, Yasui K, Tomie A, Gen Y, Terasaki K, Kitaichi T, et al. Oncogenic miR-96-5p inhibits apoptosis by targeting the caspase-9 gene in hepatocellular carcinoma. Int J Oncol. 2018;53(1):237-5.
6. 6. Xie W, Sun F, Chen L, Cao X. miR-96 promotes breast cancer metastasis by suppressing MTSS1. Oncol Lett. 2018;15(3):3464-1.
7. 7. Ress AL, Stiegelbauer V, Winter E, Schwarzenbacher D, Kiesslich T, Lax S, et al. MiR‐96‐5p influences cellular growth and is associated with poor survival in colorectal cancer patients. Mol. Carcinog. 2015;54(11):1442-50.
8. 8. Zheng A, Zhang L, Song X, Wang Y, Wei M, Jin F. Clinical implications of a novel prognostic factor AIFM3 in breast cancer patients. BMC cancer. 2019;19(1):451.

9. 9. Lewis BP, Burge CB, Bartel DP. Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell. 2005;120(1):15-0.
10. 10. Esquela-Kerscher A, Slack FJ. Oncomirs—microRNAs with a role in cancer. Nat Rev Cancer. 2006;6(4):259-9.
11. 11. Hong Y, Liang H, Wang Y, Zhang W, Zhou Y, Yu M, et al. miR-96 promotes cell proliferation, migration and invasion by targeting PTPN9 in breast cancer. Sci. Rep. 2016;6:37421.
12. 12. Yao Q, Pei Y, Zhang X, Xie B. microRNA-96 acts as a tumor suppressor gene in human osteosarcoma via target regulation of EZRIN. Life Sci. 2018;203:1-11.
13. 13. Yu N, Fu S, Liu Y, Xu Z, Liu Y, Hao J, et al. miR-96 suppresses renal cell carcinoma invasion via downregulation of Ezrin expression. J Exp Clin Cancer Res. 2015;34(1):107.
14. 14. Wu L, Pu X, Wang Q, Cao J, Xu F, Xu LI, et al. miR‑96 induces cisplatin chemoresistance in non‑small cell lung cancer cells by downregulating SAMD9. Oncol Lett. 2016;11(2):945-2.