AURKB Geni miR-34a-5p ve let-7b-5p Tarafından Hedeflendiğinden AML Hücre Proliferasyonunda Rol Oynar mı?

Abstract

Amaç: Lösemik blastların çoğalarak birikimiyle karakterize olan AML’de kemik iliğindeki normal kan hücrelerinin üretimi sekteye uğradığından hastalar anemi ve trombositopeni sorunu yaşamaktadır. AML'de yüksek oranda eksprese edildiği bildirilen Aurora kinazların gen ekspresyonu azaldığında AML'deki klinik bulguları hafifletmesi mümkün olabilir. Bu çalışmada, AURKB ile protein ekspresyonunu düzenleme potansiyeli bulunan önemli miRNA’ların ilişkisinin incelenmesi amaçlanmıştır. Metod: HL60 ve NB4 hücreleri, önemli tümor supresör miRNA’lardan olan miR-34a-5p ve let-7b-5p mimikleri ile tarnsfekte edilmiştir. Ardından WST-8 tekniğiyle proliferasyona etkileri incelenmiş ve qRT-PZR ile de AURKB gen ifadesine etkileri incelenmiştir. Bulgular: Her iki AML hücre hattında da bu miRNA’ların proliferasyonu negatif yönde regüle ettiği ve kontrol grubuna kıyasla miRNA transfekte edilen grupta Aurora kinase B (AURKB) geninin ifade seviyesini dowregüle ettikleri belirlenmiştir. Sonuç: Sonuç olarak miR-34a-5p ve let-7b-5p’nin AML hücrelerinde AURKB ifadesini düzenleyebileceği tespit edilmiştir. Bu nedenle, AML’deki aşırı hücre bölünmesi ve kötü prognozun engellenebilmesinde bu miRNA’ların terapötik bir biyomarker olarak önemli bir potansiyeli olabileceği düşünülmüştür.

Keywords

• AML
• AURKB
• miR-34a-5p
• let-7b-5p

Is the AURKB Gene Involved in Aml Cell Proliferation Since It is Targeted by miR-34a-5p and let-7b-5p?

Abstract

Objective: The production of normal blood cells in the bone marrow is interrupted in AML, which is characterized by the proliferation and accumulation of leukemic blasts. Therefore, patients experience anemia and thrombocytopenia. When gene expression of Aurora kinases, which is reported to be highly expressed in AML, decreases, it may be possible to alleviate the clinical findings in AML. In this study, it was aimed to examine the relationship of AURKB with important miRNAs that have the potential to regulate gene expression. Method: HL60 and NB4 cells were transfected with important tumor suppressor miRNAs miR-34a-5p and let-7b-5p mimics. Then, its effects on proliferation were examined with WST-8 technique and its effects on AURKB gene expression were examined with qRT-PCR. Results: It was determined that these miRNAs negatively regulated proliferation in both AML cell lines and downregulated the expression level of the Aurora kinase B (AURKB) gene in the miRNA transfected group compared to the control group. Conclusion: In conclusion, it was determined that miR-34a-5p and let-7b-5p could regulate AURKB expression in AML cells. Therefore, it was thought that these miRNAs may have an important potential as a therapeutic biomarker in preventing excessive cell division and poor prognosis in AML.

Keywords

• AML
• AURKB
• miR-34a-5p
• let-7b-5p

References

1. 1. Pelcovits A, Niroula R. Acute Myeloid Leukemia: A Review. R I Med J (2013). 2020;103(3):38-40.
2. 2. De Kouchkovsky I, Abdul-Hay M. Acute myeloid leukemia: a comprehensive review and 2016 update. Blood Cancer J. 2016;6.
3. 3. Kojima K, Konopleva M, Tsao T, Nakakuma H, Andreeff M. Concomitant inhibition of Mdm2-p53 interaction and Aurora kinases activates the p53-dependent postmitotic checkpoints and synergistically induces p53-mediated mitochondrial apoptosis along with reduced endoreduplication in acute myelogenous leukemia. Blood. 2008;112(7):2886-95.
4. 4. Carmena M, Earnshaw WC. The cellular geography of aurora kinases. Nat Rev Mol Cell Bio. 2003;4(11):842-54.
5. 5. Chieffi P, Cozzolino L, Kisslinger A, Libertini S, Staibano S, Mansueto G, et al. Aurora B expression directly correlates with prostate cancer malignancy and influence prostate cell proliferation. Prostate. 2006;66(3):326-33.
6. 6. García-Fernández E, De Diego JI, Collantes-Bellido E, Mendiola M, Prim MP, Pérez-Fernández E, et al. Aurora B kinase expression in laryngeal squamous cell carcinoma and its prognostic implications. Histopathology. 2011;58(3):368-76.
7. 7. Yang J, Ikezoe T, Nishioka C, Tasaka T, Taniguchi A, Kuwayama Y, et al. AZD1152, a novel and selective aurora B kinase inhibitor, induces growth arrest, apoptosis, and sensitization for tubulin depolymerizing agent or topoisomerase II inhibitor in human acute leukemia cells in vitro and in vivo. Blood. 2007;110(6):2034-40.
8. 8. Manfredi MG, Ecsedy JA, Meetze KA, Balani SK, Burenkova O, Chen W, et al. Antitumor activity of MLN8054, an orally active small-molecule inhibitor of Aurora A kinase. Proc Natl Acad Sci U S A. 2007;104(10):4106-11.

9. 9. Emmrich S, Katsman-Kuipers JE, Henke K, Khatib ME, Jammal R, Engeland F, et al. miR-9 is a tumor suppressor in pediatric AML with t(8;21). Leukemia. 2014;28(5):1022-32.
10. 10. Stavast CJ, van Zuijen I, Karkoulia E, Özçelik A, van Hoven-Beijen A, Leon LG, et al. The tumor suppressor MIR139 is silenced by POLR2M to promote AML oncogenesis. Leukemia. 2022;36(3):687-700.
11. 11. Xue H, Hua LM, Guo M, Luo JM. SHIP1 is targeted by miR-155 in acute myeloid leukemia. Oncol Rep. 2014;32(5):2253-9.
12. 12. 06.06.2022 [Available from: https://servers.binf.ku.dk/bloodspot/.
13. 13. 07.06.2022 [Available from: https://string-db.org/.
14. 14. Mardani R, Jafari Najaf Abadi MH, Motieian M, Taghizadeh-Boroujeni S, Bayat A, Farsinezhad A, et al. MicroRNA in leukemia: Tumor suppressors and oncogenes with prognostic potential. J Cell Physiol. 2019;234(6):8465-86.
15. 15. Turk A, Calin GA, Kunej T. MicroRNAs in Leukemias: A Clinically Annotated Compendium. Int J Mol Sci. 2022;23(7).
16. 16. 08.06.2022 [Available from: https://www.mirnet.ca/.
17. 17. 20.06.2022 [Available from: http://www.mirdb.org/.
18. 18. 21.06.2022 [Available from: www.targetscan.org.
19. 19. 10.06.2022 [Available from: http://mirwalk.umm.uni-heidelberg.de/.
20. 20. 01.09.2022 [Available from: https://sfold.wadsworth.org/starmirDB.php.
21. 21. Mäki-Jouppila JH, Pruikkonen S, Tambe MB, Aure MR, Halonen T, Salmela AL, et al. MicroRNA let-7b regulates genomic balance by targeting Aurora B kinase. Mol Oncol. 2015;9(6):1056-70.
22. 22. Suer I, Karatas OF, Yuceturk B, Yilmaz M, Guven G, Buge O, et al. Characterization of stem-like cells directly isolated from freshly resected laryngeal squamous cell carcinoma specimens. Curr Stem Cell Res Ther. 2014;9(4):347-53.
23. 23. Döhner H, Estey E, Grimwade D, Amadori S, Appelbaum FR, Büchner T, et al. Diagnosis and management of AML in adults: 2017 ELN recommendations from an international expert panel. Blood. 2017;129(4):424-47.
24. 24. Liu H. Emerging agents and regimens for AML. J Hematol Oncol. 2021;14(1):49.
25. 25. Kelly JR, Martini S, Brownlow N, Joshi D, Federico S, Jamshidi S, et al. The Aurora B specificity switch is required to protect from non-disjunction at the metaphase/anaphase transition. Nat Commun. 2020;11(1):1396.
26. 26. Keen N, Taylor S. Mitotic drivers--inhibitors of the Aurora B Kinase. Cancer Metastasis Rev. 2009;28(1-2):185-95.
27. 27. Moore AS, Blagg J, Linardopoulos S, Pearson AD. Aurora kinase inhibitors: novel small molecules with promising activity in acute myeloid and Philadelphia-positive leukemias. Leukemia. 2010;24(4):671-8.
28. 28. Qi J, Gao X, Zhong X, Zhang N, Wang R, Zhang H, et al. Selective inhibition of Aurora A and B kinases effectively induces cell cycle arrest in t(8;21) acute myeloid leukemia. Biomed Pharmacother. 2019;117:109113.
29. 29. Yang J, Ikezoe T, Nishioka C, Tasaka T, Taniguchi A, Kuwayama Y, et al. AZD1152, a novel and selective aurora B kinase inhibitor, induces growth arrest, apoptosis, and sensitization for tubulin depolymerizing agent or topoisomerase II inhibitor in human acute leukemia cells in vitro and in vivo. Blood. 2007;110(6):2034-40.
30. 30. Joshi SK, Nechiporuk T, Bottomly D, Piehowski PD, Reisz JA, Pittsenbarger J, et al. The AML microenvironment catalyzes a stepwise evolution to gilteritinib resistance. Cancer Cell. 2021;39(7):999-1014.e8.
31. 31. Hartsink-Segers SA, Zwaan CM, Exalto C, Luijendijk MW, Calvert VS, Petricoin EF, et al. Aurora kinases in childhood acute leukemia: the promise of aurora B as therapeutic target. Leukemia. 2013;27(3):560-8.
32. 32. Shi Q, Xing G, Qi M, Xing Y. Lower Serum miR-145 Predicts Poor Prognosis in Patients with Acute Myeloid Leukemia. Clin Lab. 2020;66(6).
33. 33. Huang Y, Zou Y, Lin L, Ma X, Chen H. Identification of serum miR-34a as a potential biomarker in acute myeloid leukemia. Cancer Biomark. 2018;22(4):799-805.
34. 34. Katoh M, Igarashi M, Fukuda H, Nakagama H, Katoh M. Cancer genetics and genomics of human FOX family genes. Cancer Lett. 2013;328(2):198-206.
35. 35. Peng Y, Fan JY, Xiong J, Lou Y, Zhu Y. miR-34a Enhances the Susceptibility of Gastric Cancer to Platycodin D by Targeting Survivin. Pathobiology. 2019;86(5-6):296-305.
36. 36. Zhou X, Ma X, Sun H, Li X, Cao H, Jiang Y, et al. Let-7b regulates the adriamycin resistance of chronic myelogenous leukemia by targeting AURKB in K562/ADM cells. Leuk Lymphoma. 2020;61(14):3451-9.