Abstract
Background
MicroRNAs play important roles in the pathogenesis of human diseases by regulating target gene expression in specific cells or tissues. Previously, we identified microRNA 452 (MIR452), which was specifically up-regulated in early stage human colorectal cancer (CRC) tissue.
Objective
The current study aims to identify and verify the target genes of MIR452 associated with CRC.
Methods
A luciferase reporter system was used to confirm the effect of MIR452 on ASB8, NOL8, and CDR2 expression. The expression levels of MIR452 and the target genes were evaluated by quantitative RT-PCR (qRT-PCR) and western blotting.
Results
We verified the association between MIR452 and three genes, ASB8, NOL8, and CDR2, and showed that their transcripts were down-regulated by MIR452. Up-regulated MIR452 also down-regulated ASB8, NOL8, and CDR2 mRNA and protein levels in CRC cells. CDR2 protein expression was decreased in CRC tissues compared to adjacent non-tumor tissues.
Conclusions
These results suggest that ASB8, NOL8, and CDR2 were target genes of MIR452 in CRC cells and that up-regulated MIR452 in CRC tissue regulated ASB8, NOL8, and CDR2 expression during colorectal carcinogenesis.
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Data availability
All data generated or analyzed during this study are included in this published article.
References
Alam KJ, Mo JS, Han SH, Park WC, Kim HS, Yun KJ, Chae SC (2017) MicroRNA 375 regulates proliferation and migration of colon cancer cells by suppressing the CTGF-EGFR signaling pathway. Int J Cancer 141(8):1614–1629. https://doi.org/10.1002/ijc.30861
Andresen CA, Smedegaard S, Sylvestersen KB, Charlotte S, Diego IG, Giuseppe C, Tine Kragh N, Michael LN, Amilcar FM (2014) Protein interaction screening for the ankyrin repeats and suppressor of cytokine signaling (SOCS) box (ASB) family Identify Asb11 as a novel endoplasmic reticulum resident ubiquitin ligase. J Biol Chem 289(4):2043–2054. https://doi.org/10.1074/jbc.M113.534602
Aydin Y, Kurt R, Song K, Lin D, Osman H, Youngquist B, Scott JW, Shores NJ, Thevenot P, Cohen A, Dash S (2019) Hepatic stress response in HCV infection promotes STAT3-mediated inhibition of HNF4A-miR-122 feedback loop in liver fibrosis and cancer progression. Cancers (Basel) 11:10. https://doi.org/10.7748/ns.6.24.43.s50
Balamurugan K, Luu VD, Kaufmann MR, Hofmann VS, Boysen G, Barth S, Bordoli MR, Stiehl DP, Moch H, Schraml P, Wenger RH, Camenisch G (2009) Onconeuronal cerebellar degenerationrelated antigen, Cdr2, is strongly expressed in papillary renal cell carcinoma and leads to attenuated hypoxic response. Oncogene 28:3274–3285. https://doi.org/10.1038/onc.2009.186
Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281–297. https://doi.org/10.1016/s0092-8674(04)00045-5
Braga EA, Fridman MV, Loginov VI, Dmitriev AA, Morozov SG (2019) Molecular mechanisms in clear cell renal cell carcinoma: role of miRNAs and hypermethylated miRNA genes in crucial oncogenic pathways and processes. Front Genet 10:320. https://doi.org/10.3389/fgene.2019.00320
Brennecke J, Hipfner DR, Stark A, Russell RB, Cohen SM (2003) bantam encodes a developmentally regulated microRNA that controls cell proliferation and regulates the proapoptotic gene hid in Drosophila. Cell 113:25–36. https://doi.org/10.1016/s0092-8674(03)00231-9
Chen XL, Hong LL, Wang KL, Liu X, Wang JL, Lei L, Xu ZY, Cheng XD, Ling ZQ (2019) Deregulation of CSMD1 targeted by microRNA-10b drives gastric cancer progression through the NF-κB pathway. Int J Biol Sci 15:2075–2086. https://doi.org/10.7150/ijbs.23802
Darnell RB, Albert ML (2000) Cdr2-specific CTLs are detected in the blood of all patients with paraneoplastic cerebellar degeneration analyzed. Ann Neurol 48:270–271 (PMID: 10939585)
Garzon R, Fabbri M, Cimmino A, Calin A, Croce GA (2006) MicroRNA expression and function in cancer. Trends Mol Med 12(12):580–587. https://doi.org/10.1016/j.molmed.2006.10.006
Gu S, Hou P, Liu K, Niu X, Wei B, Mao F, Xu Z (2018) NOL8, the binding protein for beta-catenin, promoted the growth and migration of prostate cancer cells. Chem Biol Interact 294:40–47. https://doi.org/10.1016/j.cbi.2018.08.019
Han SH, Mo JS, Park WC, Chae SC (2019) Reduced microRNA 375 in colorectal cancer upregulates metadherin-mediated signaling. World J Gastroenterol 25(44):6495–6507 (PMID: 1802830)
Jinawath N, Furukawa Y, Nakamura Y (2004) Identification of NOL8, a nucleolar protein containing an RNA recognition motif (RRM), which was overexpressed in diffuse-type gastric cancer. Cancer Sci 95(5):430–435. https://doi.org/10.1111/j.1349-7006.2004.tb03227.x
Kulkarni P, Dasgupta P, Bhat NS, Shahryari V, Shiina M, Hashimoto Y, Majid S, Deng G, Saini S, Tabatabai ZL, Yamamura S, Tanaka Y, Dahiya R (2018) Elevated miR-182-5p associates with renal cancer cell mitotic arrest through diminished MALAT-1 expression. Mol Cancer Res 16:1750–1760. https://doi.org/10.1158/1541-7786.MCR-17-0762
Liu YZ, Li JJ, Zhang FG, Wenxin Q, Genfu Y, Xianghuo H, Peng X, Chao G, Dafang W, Jianren G (2003) Molecular cloning and characterization of the human ASB-8 gene encoding a novel member of ankyrin repeat and SOCS box containing protein family. Biochem Biophys Res Commun 300:972–979. https://doi.org/10.1016/s0006-291x(02)02971-6
Liu L, Chen K, Wu J, Shi L, Hu B, Cheng S, Li M, Song L (2013) Downregulation of miR-452 promotes stem-like traits and tumorigenicity of gliomas. Clin Cancer Res 19:3429–3438. https://doi.org/10.1158/1078-0432.CCR-12-3794
Lu J, Getz G, Miska EA, Alvarez-Saavedra E, Lamb J, Peck D, Sweet-Cordero A, Ebert BL, Mak RH, Ferrando AA, Downing JR, Jacks T, Horvitz HR, Golub TR (2005) MicroRNA expression profiles classify human cancers. Nature 435:834–838. https://doi.org/10.1038/nature03702
Lu Z, Li X, Xu Y, Chen M, Chen W, Chen T, Tang Q, He Z (2019) microRNA-17 functions as an oncogene by downregulating Smad3 expression in hepatocellular carcinoma. Cell Death Dis 10:723. https://doi.org/10.1038/s41419-019-1960-z
Ma L, Teruya-Feldstein J, Weinberg RA (2007) Tumour invasion and metastasis initiated by microRNA-10b in breast cancer. Nature 449:682–688. https://doi.org/10.1038/nature06174
Mo JS, Alam KJ, Kang IH, Park WC, Seo GS, Choi SC, Kim HS, Moon HB, Yun KJ, Chae SC (2015) MicroRNA 196B regulates FAS-mediated apoptosis in colorectal cancer cells. Oncotarget 6:2843–2855. https://doi.org/10.18632/oncotarget.3066
Mo JS, Alam KJ, Kim HS, Lee YM, Yun KJ, Chae SC (2016) MicroRNA 429 regulates mucin gene expression and secretion in murine model of colitis. J Crohns Colitis 10:850–859. https://doi.org/10.1093/ecco-jcc/jjw033
Mo JS, Park WC, Choi SC, Yun KJ, Chae SC (2019) MicroRNA 452 regulates cell proliferation, cell migration, and angiogenesis in colorectal cancer by suppressing VEGFA expression. Cancers 11(10):1613. https://doi.org/10.3390/cancers11101613
O’Donvan KJ, Diedler J, Couture GC, Fak JJ, Darnell RB (2010) The onconeural antigen cdr2 is a novel APC/C target that acts in mitosis to regulate c-myc target genes in mammalian tumor cells. PLoS One 5:e10045. https://doi.org/10.1371/journal.pone.0010045
Okano HJ, Park WY, Corradi JP, Darnell RB (1999) The cytoplasmic Purkinje onconeural antigen cdr2 down-regulates c-Myc function: implications for neuronal and tumor cell survival. Genes Dev 13:2087–2097. https://doi.org/10.1101/gad.13.16.2087
Sakai K, Kitagawa Y, Saiki S, Saiki M, Hirose G (2004) Effect of a paraneoplastic cerebellar degeneration-associated neural protein on B-myb promoter activity. Neurobiol Dis 15:529–533. https://doi.org/10.1016/j.nbd.2003.11.003
Sekiguchi T, Todaka Y, Wang Y, Hirose E, Nakashima N, Nishimoto T (2004) A novel human nucleolar protein, Nop132, binds to the G proteins, RRAG A/C/D. J Biol Chem 279(9):8343–8350. https://doi.org/10.1074/jbc.M305935200
Sekiguchi T, Hayano T, Yanagida M, Takahashi N, Nishimoto T (2006) NOP132 is required for proper nucleolus localization of DEAD-box RNA helicase DDX47. Nucleic Acids Res 34(16):4593–4608. https://doi.org/10.1093/nar/gkl603
Serocki M, Bartoszewska S, Janaszak-Jasiecka A, Ochocka RJ, Collawn JF, Bartoszewski R (2018) miRNAs regulate the HIF switch during hypoxia: a novel therapeutic target. Angiogenesis 21:183–202. https://doi.org/10.1007/s10456-018-9600-2
Shan J, Al-Muftah MA, Al-Kowari MK, Abuaqel SWJ, Al-Rumaihi K, Al-Bozom I, Li P, Chouchane L (2019) Targeting Wnt/EZH2/microRNA-708 signaling pathway inhibits neuroendocrine differentiation in prostate cancer. Cell Death Discov 5:139. https://doi.org/10.1038/s41420-019-0218-y
Storstein A, Vedeler CA (2007) Paraneoplastic neurological syndromes and onconeural antibodies: clinical and immunological aspects. Adv Clin Chem 44:143–185. https://doi.org/10.1016/s0065-2423(07)44005-7
Su H, Yang JR, Xu T, Huang J, Li Xu, Yuan Y, Zhuang SM (2009) MicroRNA-101, down-regulated in hepatocellular carcinoma, promotes apoptosis and suppresses tumorigenicity. Cancer Res 69:1135–1142. https://doi.org/10.1158/0008-5472.CAN-08-2886
Takanaga H, Mukai H, Shibata H, Toshimori M, Ono Y (1998) PKN interacts with a paraneoplastic cerebellar degenerationassociated antigen, which is a potential transcription factor. Exp Cell Res 241:363–372. https://doi.org/10.1006/excr.1998.4060
Totland C, Aarskog NK, Eichler TW, Haugen M, Nøstbakken JK, Monstad SE, Salvesen HB, Mørk S, Haukanes BI, Vedeler CA (2011) CDR2 antigen and Yo antibodies. Cancer Immunol Immunother 60(2):283–289. https://doi.org/10.1007/s00262-010-0943-9
Van Schooneveld E, Wouters MC, Van der Auwera I, Peeters DJ, Wildier H, Van Dam PA, Vergote I, Vermeulen PB, Dirix LY, Van Laere SJ (2012) Expression profiling of cancerous and normal breast tissues identifies microRNAs that are differentially expressed in serum from patients with (metastatic) breast cancer and healthy volunteers. Breast Cancer Res 14:R34. https://doi.org/10.1186/bcr3127
Veerla S, Lindgren D, Kvist A, Frigyesi A, Staaf J, Persson H, Liedberg F, Chebil G, Gudjonsson S, Borg A et al (2009) MiRNA expression in urothelial carcinomas: important roles of miR-10a, miR-222, miR-125b, miR-7 and miR-452 for tumor stage and metastasis, and frequent homozygous losses of miR-31. Int J Cancer 124:2236–2242. https://doi.org/10.1002/ijc.24183
Wang Y, Toh HC, Chow P, Chung AYF, Meyers DJ, Cole PA, Ooi LL, Lee CG (2012) MicroRNA-224 is upregulated in hepatocellular carcinoma through epigenetic mechanisms. Faseb J 26:3032–3041. https://doi.org/10.1096/fj.11-201855
Wang YF, Ao X, Liu Y, Ding D, Jiao WJ, Yu Z, Zhai WX, Dong SH, He YQ, Guo H, Wang JX (2019) MicroRNA-608 promotes apoptosis in non-small cell lung cancer cells treated with doxorubicin through the inhibition of TFAP4. Front Genet 10:809. https://doi.org/10.3389/fgene.2019.00809
Zhang Z, Pi J, Zou D, Wang X, Xu J, Yu S, Zhang T, Li F, Zhang X, Zhao H, Wang F, Dong W, Ma Y, Jia YJ (2019a) microRNA arm-imbalance in part from complementary targets mediated decay promotes gastric cancer progression. Nat Commun 10:4397. https://doi.org/10.1038/s41467-019-12292-5
Zhang WC, Wells JM, Chow KH, Huang H, Yuan M, Saxena T, Melnick MA, Politi K, Asara JM, Costa DB (2019b) miR-147b-mediated TCA cycle dysfunction and pseudohypoxia initiate drug tolerance to EGFR inhibitors in lung adenocarcinoma. Nat Metab 1:460–474. https://doi.org/10.1038/s42255-019-0052-9
Zheng Q, Sheng Q, Jiang C, Shu J, Chen J, Nie Z, Lv Z, Zhang Y (2014) MicroRNA-452 promotes tumorigenesis in hepatocellular carcinoma by targeting cyclin-dependent kinase inhibitor 1B. Mol Cell Biochem 389:187–195. https://doi.org/10.1007/s11010-013-1940-z
Acknowledgements
The biospecimens for this study were provided by the Biobank of Wonkwang University Hospital, a member of the National Biobank of Korea, which is supported by the Ministry of Health and Welfare. This research was supported by Wonkwang University in 2018.
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This research was supported by Wonkwang University in 2018.
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J-S Mo performed the experiments and analyzed the data, and S-C Chae conceived, designed the experiments, analyzed the data and wrote the manuscript.
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Ji-Su Mo and Soo-Cheon Chae declare that they have no conflicts of interests.
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The study (WKIRB-201703-BR-010) was reviewed and approved by the review board of Wonkwang University, Republic of Korea. This study did not use animal's subjects.
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Mo, JS., Chae, SC. MicroRNA 452 regulates ASB8, NOL8, and CDR2 expression in colorectal cancer cells. Genes Genom 43, 33–41 (2021). https://doi.org/10.1007/s13258-020-01016-5
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DOI: https://doi.org/10.1007/s13258-020-01016-5