MEME KANSERİNDE MİTOMİR’LERİN TANIMLANMASI: İN VİTRO ÇALIŞMA İLE ROLLERİNİN GÖSTERİLMESİ

Abstract

Amaç: Kadınlarda meme kanseri %5 oranında genetik yatkınlıkla ilişkilidir. Mitokondri, metabolizma, hücre ölümü ve inflamasyon gibi önemli hücresel olaylarda rol oynar. Son çalışmalar, olgun miRNA’ların yanı sıra pre-miRNA’ların mitokondride bulunduğunu göstermişlerdir. Bu çalışmada meme kanseri hücrelerinde mitomiR’lerin varlığını ortaya koymayı amaçladık. Gereç ve Yöntem: MACS yöntemi kullanılarak mitokondriyal fraksiyonlar hazırlandı ve küçük RNA dizilimi yapıldı. Bulgular: Mitokondriyal genoma hizalanmış bilinen ve yeni mitomiR dizilerini belirledik. Sonuç: Yeni mitomiR’lerin tanımlanması literatüre önemli katkılar sağlayabilir ve mitomiR biyogenezi moleküler mekanizmalar ile aydınlatılabilir.

Keywords

• meme kanseri
• mitomiR
• MACS
• küçük RNA dizileme

Project Number

2014SBE011

EXPLORING MITOMIRS IN BREAST CANCER: AN IN-VITRO STUDY OF THEIR EMERGING ROLES

Abstract

Objective: Breast cancer is associated with a 5% genetic predisposition in women. Mitochondria play a role in important cellular events such as metabolism, cell death, and inflammation. Recent studies have highlighted precursor micro-RNA (pre-miRNA) to be located in mitochondria as well as mature miRNA. This study aims to reveal the occurrence of mitochondrial miRNA (mitomiR) in breast cancer cells. Materials and Methods: The study has prepared the mitochondrial fractions using the magnetic-activated cell sorting (MACS) method and performed small RNA (sRNA) sequencing. Results: The study has identified known and novel mitomiR sequences aligned to the mitochondrial genome. Conclusion: Identifying new mitomiRs can provide significant contributions and illuminate the molecular mechanism underlying mitomiR biogenesis.

Keywords

• mitomiRs
• MACS
• breast cancer
• small RNA sequencing

Supporting Institution

Pamukkale Üniversitesi BAP

Project Number

2014SBE011

Thanks

We want to thank Prof. Dr. Gülseren Bağcı for her laboratory support and scientific comments, Assoc.Prof.Dr. Onur Tokgun for contributing with his valuable scientific support and Msc.Serap Kurt for laboratory support.

References

1. 1. Chatterjee A, Mambo E, Sidransky D. Mitochondrial DNA mutations in human cancer. Oncogene 2006;25(34): 663-74. [CrossRef] google scholar
2. 2. Plak K, Czarnecka AM, Krawczyk T, Golik P, Bartnik E. Breast cancer as a mitochondrial disorder (Review) Oncol Rep 2009;21(4):845-51. [CrossRef] google scholar
3. 3. Kushnareva Y, Newmeyer DD. Bioenergetics and cell death. Ann N Y Acad Sci 2010;1201:50-7. [CrossRef] google scholar
4. 4. Bienervota-Vasku J, Sana J, Slaby O. The role of miRNAs in mitochondria in cancer. Cancer Letters 2013;336:1-7. [CrossRef] google scholar
5. 5. Dimauro S. A history of mitochondrial diseases. J Inherit Metab Dis 2011;34:261-76. [CrossRef] google scholar
6. 6. Zorzano A, Hernandez-Alvarez MI, Palacin M, Mingrone G. Alterations in the mitochondrial regulatory pathways constituted by the nuclear co-factors PGC-1alpha or PGC-1beta and Mitofusin 2 in skeletal muscle in type 2 diabetes. Biochim Biophys Acta 2010;1797:1028-33. [CrossRef] google scholar
7. 7. Peralta S, Wang X, Moraes CT. Mitochondrial transcription: lessons from mouse models. Biochim Biophys Acta 2012;1819:961-9. [CrossRef] google scholar
8. 8. Boore JL. Animal mitochondrial genomes. Nucleic Acids Res 1999;27(8):1767-80. [CrossRef] google scholar

9. 9. Bertram JS. The molecular biology of the cancer. Mol Aspects Med 2000;21(6):167-223. [CrossRef] google scholar
10. 10. Kim S, Koh H. Role of FOXO transcription factors in crosstalk between mitochondria and the nucleus. J Bioenerg Biomembr 2017;49(4):335-41. [CrossRef] google scholar
11. 11. Sripada L, Tomar D, Singh R. Mitochondria: one of the destinations of miRNAs. Mitochondrion 2012;12:593-9. [CrossRef] google scholar
12. 12. Mercer TR, Neph S, Dinger ME, Crawford J, Smith MA, Shearwood AM, et al. The human mitochondrial transcriptome. Cell 2011;146 (4):645-58. [CrossRef] google scholar
13. 13. Entelis NS, Kolesnikova OA, Martin RP, Tarassov IA. RNA delivery into mitochondria. Adv Drug Deliv Rev 2001;49(1-2):199-215. [CrossRef] google scholar
14. 14. Puranam RS, Attardi G. The RNase P associated with Hela cell mitochondria contains an essential RNA component identical in sequence to that of the nuclear RNase P. Mol Cell Biol 2001;21(2):548-61. [CrossRef] google scholar
15. 15. Alfonzo JD, Soll D. Mitochondrial tRNA import—the challenge to understand has just begun. Biol Chem 2009;390(8):717-22. [CrossRef] google scholar
16. 16. Sripada L, Tomar D, Prajapati P, Singh R, Singh AK, Singh R. Systematic analysis of small RNAs associated with human mitochondria by deep sequencing: detailed analysis of mitochondrial associated miRNA. Plos One 2012;7(9):e44873. [CrossRef] google scholar
17. 17. Wang WX. Role of mitochondria in regulating microRNA activitiy and its relevence to the central nerveous system. Neural Regen Res 2015;10(7):1026-28. [CrossRef] google scholar
18. 18. Imanishi H, Hattori K, Wada R, Ishikawa K, Fukuda S, Takenaga K, et al. Mitochondrial DNA mutations regulate metastasis of human breast cancer cells. Plos One 2011;6(8):e23401. [CrossRef] google scholar
19. 19. Bandiera S, Rüberg S, Girard M, Cagnard N, Hanein S, Chrétien D, et al. Nuclear outsourcing of RNA interference components to human mitochondria. Plos One 2011;6(6):e20746. [CrossRef] google scholar
20. 20. Wang X, Song C, Zhou X, Han X, Li J, Wang Z. Mitochondria Associated MicroRNA Expression Profiling of Heart Failure. BioMed Res Int 2017;2017:4042509. [CrossRef] google scholar
21. 21. Hornig-Do HT, Günther G, Bust M, Lehnartz P, Bosio A, Wiesner RJ. Isolation of functional pure mitochondria by superparamagnetic microbeads. Anal Biochem 2009;389(1):1-5. [CrossRef] google scholar
22. 22. Chintha R, Kaipa PR, Sekhar N, Hasan Q. Mitochondria and tumors: a new perspective. Indian J Cancer 2013;50(3):206-13. [CrossRef] google scholar
23. 23. Bandiera S, Matégot R, Girard M, Demongeot J, Henrion-Caude A. MitomiRs delineating the intracellular localization of microRNAs at mitochondria. Free Radic Biol Med 2013;64:12-9. [CrossRef] google scholar
24. 24. Tamaddon G, Geramizadeh B, Karimi MH, Mowla SJ, Abroun S. miR-4284 and miR-4484 as Putative Biomarkers for Diffuse Large B-Cell Lymphoma. Iranian J Med Sci 2016;41(4):334-9. google scholar
25. 25. Li J, Zhang X, Sejas DP, Bagby GC, Pang Q. Hypoxia-induced nucleophosmin protects cell death through inhibition of p53. J Biol Chem 2004;279(40):41275-9. [CrossRef] google scholar
26. 26. Dhar SK, St Clair DK. Nucleophosmin blocks mitochondrial localization of p53 and apoptosis. J Biol Chem 2009;284(24):16409-18. [CrossRef] google scholar