MiR-490-5p Functions as an OncomiR in Breast Cancer by Targeting NFATc4

Document Type : Research Article


Department of Laboratory Sciences, Kashan Branch, Islamic Azad University, Kashan, Iran


Breast cancer is a serious health problem worldwide in women. MicroRNAs are small non-coding RNAs of 18–25 nucleotides in length that post-transcriptionally modulate gene expression. MiR-490 has been reported as a tumor suppressor and oncomiR microRNA in breast cancer with two separate targets, NFAT and Rho. NFAT is one of the targets for miR-490 but the relationship between hsa-miR-490 and NFATc3, NFATc4 are not clear yet. Except for NFAT5, the other members of NFAT are activated by Ca2+ influx in the cell, either via the PLC-γ pathway or via store-operated Ca2+ entry, typically in T lymphoid cells. In a cross-sectional comparative study, peripheral blood samples were collected from 30 subjects with breast cancer and 30 healthy individuals as a control group. Gene expression analysis of peripheral blood mononuclear cells (PBMCs) was performed using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to study the NFATc3, NFATc4, and hsa-miR-490-5p gene expression alterations.   As per the obtained results, a significant decrease was observed in the expression level of NFATc4 (p <0.05), while hsa-miR-490-5p expression found to be elevated in PBMCs of breast cancer patient (p <0.05).  Expression changes were not significant for NFATc3 gene (P>0.05). Taken together findings of this study indicated that serum hsa-miR-490-5p acts as an oncomiR by direct targeting the NFATc4.


Aminisepehr F, Babaei E, Hosseinpour Feizi MA. 2018. Study of the expression of miR-4270 in plasma of patients with breast invasive ductal carcinoma. J Genet Resour 4:85-89.
Becker TS, Moreira MA, Lima LA, De Oliveira ELC, Freitas‐Júnior R. 2010. Pilomatrixoma mimicking breast cancer in man. Breast J 16:89-91.
Benedito AB, Lehtinen M, Massol R, Lopes UG, Kirchhausen T, Rao A, Bonni A. 2005. The transcription factor NFAT3 mediates neuronal survival. J Biol Chem 280:2818-2825.
Cai H, Hu B, Ji L, Ruan X, Zheng Z. 2018. Hsa_circ_0103809 promotes cell proliferation and inhibits apoptosis in hepatocellular carcinoma by targeting miR-490-5p/SOX2 signaling pathway. Am J Transl Res 10:1690-1702.
Chen K, Zeng J, Tang K, Xiao H, Hu J, Huang C. 2016. miR4905p suppresses tumour growth in renal cell carcinoma through targeting PIK3CA. Biol Cell 108: 41-50.
Chen S, Chen X, Xiu Y-L, Sun K-X, Zong Z-H, Zhao Y. 2014. microRNA 490-3P enhances the drug-resistance of human ovarian cancer cells. J Ovarian Res 7:84. doi: 10.1186/s13048-014-0084-4.
Chen W, Ye L, Wen D, Chen F. 2019. miR-490-5p inhibits hepatocellular carcinoma cell proliferation, migration and invasion by directly regulating ROBO1. Pathol Oncol Res 25:1-9.
Chung Y-H, Li S-C, Kao Y-H, Luo H-L, Cheng Y-T, and Lin P-R. 2017. MiR-30a-5p inhibits epithelial-to-mesenchymal transition and upregulates expression of tight junction protein claudin-5 in human upper tract urothelial carcinoma cells. Int J Mol Sci 18: 1826. doi:10.3390/ijms18081826.
Cockerill G, Bert A, Ryan G, Gamble J, Vadas M, Cockerill P. 1995. Regulation of granulocyte-macrophage colony-stimulating factor and e-selectin expression in endothelial cells by cyclosporin A and the T-cell transcription factor NFAT. Blood 86(7):2689-2698.
Ghosn B, Benisi-Kohansal S, Ebrahimpour-Koujan S, Azadbakht L, Esmaillzadeh A. 2020. Association between healthy lifestyle score and breast cancer. Nutr J 19:1-11.
González-Patiño D, Villuendas-Rey Y, Argüelles-Cruz A-J, Karray F. 2019. A novel bio-inspired method for early diagnosis of breast cancer through mammographic image analysis. Appl Sci 9:4492. doi:10.3390/app9214492.
Graef IA, Chen F, Chen L, Kuo A, Crabtree GR. 2001. Signals transduced by Ca2+/calcineurin and NFATc3/c4 pattern the developing vasculature. Cell 105: 863-875.
Gu H, Yang T, Fu S, Chen X, Guo L, Ni Y. 2014. MicroRNA-490-3p inhibits proliferation of A549 lung cancer cells by targeting CCND1. Biochem Biophys Res Commun 444:104-108.
Ho SN, Thomas DJ, Timmerman LA, Li X, Francke U, Crabtree GR. 1995. NFATc3, a lymphoid-specific NFATc family member that is calcium-regulated and exhibits distinct DNA binding specificity. J Biol Chem 270: 19898-19907.
Hoey T, Sun Y-L, Williamson K, Xu X. 1995. Isolation of two new members of the NF-AT gene family and functional characterization of the NF-AT proteins. Immunity 2: 461-472.
Jiang L, Cao S. 2020. Role of microRNA‐26a in cartilage injury and chondrocyte proliferation and apoptosis in rheumatoid arthritis rats by regulating expression of CTGF. J Cell Physiol 235: 979-992.
Jinnin M, Medici D, Park L, Limaye N, Liu Y, Boscolo E. 2008. Suppressed NFAT-dependent VEGFR1 expression and constitutive VEGFR2 signaling in infantile hemangioma. Nat Med 14: 1236-1246.
Kalivendi SV, Konorev EA, Cunningham S, Vanamala SK, Kaji EH, Joseph J, Kalyanaraman B. 2005. Doxorubicin activates nuclear factor of activated T-lymphocytes and Fas ligand transcription: role of mitochondrial reactive oxygen species and calcium. Biochem J 389:527-539.
Key TJ, Verkasalo PK, Banks E. 2001. Epidemiology of breast cancer. Lancet Oncol 2:133-140.
Lewis BP, Burge CB, Bartel DP. 2005. Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 120:15-20.
Liang Z, Wang X, Xu X, Xie B, Ji A, Meng S. 2017. MicroRNA-608 inhibit proliferation of bladder cancer via AKT/FOXO3a signaling pathway. Mol Cancer 16: 96. doi: 10.1186/s12943-017-0664-1.
Lomasney JW, Cheng H-F, Kobayashi M, King K. 2012. Structural basis for calcium and phosphatidylserine regulation of phospholipase C δ1. Biochemistry 51: 2246-2257.
López-Rodríguez C, Antos CL, Shelton JM, Richardson JA, Lin F, Novobrantseva TI. 2004. Loss of NFAT5 results in renal atrophy and lack of tonicity-responsive gene expression. Proc Natl Acad Sci 101: 2392-2397.
Luo C, Shaw K, Raghavan A, Aramburu J, Garcia-Cozar F, Perrino BA. 1996. Interaction of calcineurin with a domain of the transcription factor NFAT1 that controls nuclear import. Proc Natl Acad Sci 93: 8907-8912.
Maillet M, Davis J, Auger-Messier M, York A, Osinska H, Piquereau J. 2010. Heart-specific deletion of CnB1 reveals multiple mechanisms whereby calcineurin regulates cardiac growth and function. J Biol Chem 285: 6716-6724.
Mancini M, Toker A. 2009. NFAT proteins: emerging roles in cancer progression. Nat Rev Cancer 9: 810-820.
Mao XP, Zhang LS, Huang B, Zhou SY, Liao J, Chen LW. 2015. Mir-135a enhances cellular proliferation through post-transcriptionally regulating PHLPP2 and FOXO1 in human bladder cancer. J Transl Med 13: 86. doi:10.1186/s12967-015-0438-8.
Mark M, Ghyselinck NB, Chambon P. 2006. Function of retinoid nuclear receptors: lessons from genetic and pharmacological dissections of the retinoic acid signaling pathway during mouse embryogenesis. Annu Rev Pharmacol Toxicol 46: 451-480.
Nikfarjam A, Pornour M, Sohrabi M, Vaseghi H. 2019. Correlation between Expression of hsa-miR-490-5p and NFAT5 in Peripheral Blood Mononuclear Cell Obtained from Breast Cancer Patients. J Genet Resour 5: 31-37.
Nishida M, Sugimoto K, Hara Y, Mori E, Morii T, Kurosaki T. 2003. Amplification of receptor signaling by Ca2+ entry‐mediated translocation and activation of PLCγ2 in B lymphocytes. EMBO J 22: 4677-4688.
Nishimura K, Sano M, Ohtaka M, Furuta B, Umemura Y, Nakajima Y. 2011. Development of defective and persistent Sendai virus vector a unique gene delivery/expression system ideal for cell reprogramming. J Biol Chem 286: 4760-4771.
Paluch-Shimon S, Cardoso F, Partridge AH, Abulkhair O, Azim JH, Bianchi-Micheli G, et al. 2020. ESO-ESMO 4th International Consensus Guidelines for Breast Cancer in Young Women (BCY4). Ann Oncol S0923-7534(20)36363-8. doi: 10.1016/j.annonc.2020.03.284.
Pan M-G, Xiong Y, Chen F. 2013. NFAT gene family in inflammation and cancer. Curr Mol Med 13: 543-554.
Pilevarzadeh M, Amirshahi M, Afsargharehbagh R, Rafiemanesh H, Hashemi SM, Balouchi A. 2019. Global prevalence of depression among breast cancer patients: a systematic review and meta-analysis. Breast Cancer Res Treat 176(3):519-533.
Qu M, Li L, Zheng W. 2017. Reduced miR-490-3p expression is associated with poor prognosis of Helicobacter pylori induced gastric cancer. Eur Rev Med Pharmacol Sci 21:3384-3388.
Rao A, Luo C, Hogan PG. 1997. Transcription factors of the NFAT family: regulation and function. Annu Rev Immunol 15: 707-747.
Schulz RA, Yutzey KE. 2004. Calcineurin signaling and NFAT activation in cardiovascular and skeletal muscle development. Dev Biol 266: 1-16.
Shimomura A, Shiino S, Kawauchi J, Takizawa S, Sakamoto H, Matsuzaki J, Ochiya T. 2016. Novel combination of serum microRNA for detecting breast cancer in the early stage. Cancer Sci 107: 326-334.
Siamakpour-Reihani S, Caster J, Nepal DB, Courtwright A, Hilliard E, Usary J. 2011. The role of calcineurin/NFAT in SFRP2 induced angiogenesis-a rationale for breast cancer treatment with the calcineurin inhibitor tacrolimus. PLoS One 6(6):e20412. doi: 10.1371/journal.pone.0020412.
Siegel RL, Miller KD, Jemal A. 2019. Cancer statistics, 2019. CA Cancer J Clin 69:7-34.
Tafrihi M, Hasheminasab E. 2019. MiRNAs: Biology, Biogenesis, Their Web-based Tools, and Databases. MicroRNA 8:4-27.
Vashishta A, Habas A, Pruunsild P, Zheng J-J, Timmusk T, Hetman M. 2009. Nuclear factor of activated T-cells isoform c4 (NFATc4/NFAT3) as a mediator of antiapoptotic transcription in NMDA receptor-stimulated cortical neurons. J Neurosci 29:15331-15340.
Wang J, Zhang X, Yao H, Le Y, Zhou W, Li J, Li X. 2020. MiR-490-5p functions as tumor suppressor in childhood neuroblastoma by targeting MYEOV. Hum Cell 33:261-271.
Wijayabahu AT, Egan KM, Yaghjyan L. Uterine cancer in breast cancer survivors: a systematic review. Breast Cancer Research and Treatment. 2020:1-19.
Wu L, Zhang M, Qi L, Zu X, Li Y, Liu L. 2019. ERα-mediated alterations in circ-0023642 and miR-490-5p signaling suppress bladder cancer invasion. Cell Death Dis 10: 1-11.
Xu X, Chen R, Li Z, Huang N, Wu X, Li S, Wu S. 2015. MicroRNA-490-3p inhibits colorectal cancer metastasis by targeting TGFβR1. BMC Cancer 15:1023. doi: 10.1186/s12885-015-2032-0.
Yang Z, Hao J, Hu ZM. 2015. MicroRNA expression profiles in human adipose-derived stem cells during chondrogenic differentiation. Int J Mol Med 35:579-586.