Abstract
MicroRNAs (miRNAs) are important gene regulators whose dysregulations can be involved in tumorigenesis. β-catenin, the main agent in the Wnt/β-catenin pathway, controls various genes and its over-expression has been discovered in different kinds of cancers including Hepatocellular Carcinoma (HCC). Extensive research demonstrated that the Wnt signaling is one of the major affected pathways in HCC. This study aimed to find miRNA targeting β-catenin gene by bioinformatic approaches and confirm this correlation to propose new therapeutic targets for HCC. Prediction of miRNAs targeting 3′-Untranslated Regions (UTR) of β-catenin mRNA, were done using different types of credible bioinformatic databases. The luciferase assay was also recruited for further confirmation of the bioinformatic predictions. In the first step, the expression of β-catenin was assessed in the HepG2 cell line by real-time PCR technique. Next, transduction of HepG2 cells were done by lentiviral vectors containing the desired miRNA. Then, the expression level of miRNA and the β-catenin gene were evaluated. Based on the results obtained from different bioinformatic databases, miR-214 was selected as the potential miRNA with the highest probability in targeting β-catenin. Furthermore, Luciferase assay results confirmed the accuracy of our bioinformatic prediction. In line with our hypothesis, after the overexpression of miR-214 in HepG2 cells, β-catenin gene expression was reduced significantly. Gathered results indicate the miRNAs role in the down-regulation of their target genes. Hence, the results propose that miR-214 can prevent HCC development by suppressing β-catenin and may supply a newfound approach towards HCC therapy in humans.
Similar content being viewed by others
References
Desai A, Sandhu S, Lai JP, Sandhu DS (2019) Hepatocellular carcinoma in non-cirrhotic liver: a comprehensive review. World J Hepatol 11(1):1–18
Rawla P, Sunkara T, Muralidharan P, Raj JP (2018) Update in global trends and aetiology of hepatocellular carcinoma. Contemp Oncol (Pozn) 22(3):141–150
Scaggiante B, Kazemi M, Pozzato G, Dapas B, Farra R, Grassi M, Zanconati F, Grassi G (2014) Novel hepatocellular carcinoma molecules with prognostic and therapeutic potentials. World J Gastroenterol WJG 20(5):1268
Wang CY, Li S (2019) Clinical characteristics and prognosis of 2887 patients with hepatocellular carcinoma: a single center 14 years experience from China. Medicine (Baltim) 98(4):e14070
Greene CM, Varley RB, Lawless MW (2013) MicroRNAs and liver cancer associated with iron overload: therapeutic targets unravelled. World J Gastroenterol WJG 19(32):5212
Khare S, Zhang Q, Ibdah JA (2013) Epigenetics of hepatocellular carcinoma: role of microRNA. World J Gastroenterol WJG 19(33):5439
Liu Y, Ding Y, Huang J, Wang S, Ni W, Guan J, Li Q, Zhang Y, Ding Y, Chen B (2014) MiR-141 suppresses the migration and invasion of HCC cells by targeting Tiam1. PLoS ONE 9(2):e88393
Lobry C, Oh P, Mansour MR, Look AT, Aifantis I (2014) Notch signaling: switching an oncogene to a tumor suppressor. Blood 123(16):2451–2459
Khalaf AM, Fuentes D, Morshid AI, Burke MR, Kaseb AO, Hassan M, Hazle JD, Elsayes KM (2018) Role of Wnt/beta-catenin signaling in hepatocellular carcinoma, pathogenesis, and clinical significance. J Hepatocell Carcinoma 5:61–73
Moeini A, Cornellà H, Villanueva A (2012) Emerging signaling pathways in hepatocellular carcinoma. Liver Cancer 1(2):83–93
Kim JW (2009) MicroRNA expression, survival, and response to interferon in liver cancer. N Engl J Med 361:1437–1447
Joo M, Lee HK, Kang YK (2003) Expression of-catenin in hepatocellular carcinoma in relation to tumor cell proliferation and cyclin D1 expression. J Korean Med Sci 18:211–217
Mohammadi-Yeganeh S, Paryan M, Samiee SM, Soleimani M, Arefian E, Azadmanesh K, Mostafavi E, Mahdian R, Karimipoor M (2013) Development of a robust, low cost stem-loop real-time quantification PCR technique for miRNA expression analysis. Mol Biol Rep 40(5):3665–3674
Tricoli L, Niture S, Chimeh U, Kumar D (2019) Role of microRNAs in the development of hepatocellular carcinoma and acquired drug resistance. Front Biosci (Landmark Ed) 24:545–554
Wang J, Lu L, Luo Z, Li W, Lu Y, Tang Q, Pu J (2019) miR-383 inhibits cell growth and promotes cell apoptosis in hepatocellular carcinoma by targeting IL-17 via STAT3 signaling pathway. Biomed Pharmacother 120:109551
Ahsani Z, Mohammadi-Yeganeh S, Kia V, Karimkhanloo H, Zarghami N, Paryan M (2017) WNT1 gene from WNT signaling pathway is a direct target of miR-122 in hepatocellular carcinoma. Appl Biochem Biotechnol 181(3):884–897
Karimkhanloo H, Mohammadi-Yeganeh S, Ahsani Z, Paryan M (2017) Bioinformatics prediction and experimental validation of microRNA-20a targeting Cyclin D1 in hepatocellular carcinoma. Tumor Biol 39(4):1010428317698361
Zhu Y, Xie J, Sun H (2019) Three miRNAs cooperate with host genes involved in human cardiovascular disease. Hum Genomics 13(1):40
Karimkhanloo H, Paryan M, Darabi M, Mohamadi-Yeganeh S, Ahsani Z (2016) Bioinformatic investigation of mirnas involved in the regulation of beta-catenin and cyclin D1 expression. Int J Adv Biotechnol Res 7(2):701–709
Friedman RC, Farh KK-H, Burge CB, Bartel DP (2009) Most mammalian mRNAs are conserved targets of microRNAs. Genome Res 19(1):92–105
Dweep H, Sticht C, Pandey P, Gretz N (2011) miRWalk–database: prediction of possible miRNA binding sites by “walking” the genes of three genomes. J Biomed Inform 44(5):839–847
Paryan M, Mohammadi-Yeganeh S, Samiee SM, Soleimani M, Arefian E, Azadmanesh K, Poopak B, Mostafavi E, Karimipoor M, Mahdian R (2013) Investigation of deregulated genes of Notch signaling pathway in human T cell acute lymphoblastic leukemia cell lines and clinical samples. Mol Biol Rep 40(10):5531–5540
Renard C-A, Labalette C, Armengol C, Cougot D, Wei Y, Cairo S, Pineau P, Neuveut C, de Reyniès A, Dejean A (2007) Tbx3 is a downstream target of the Wnt/β-catenin pathway and a critical mediator of β-catenin survival functions in liver cancer. Cancer Res 67(3):901–910
Cui J, Zhou X, Liu Y, Tang Z, Romeih M (2003) Wnt signaling in hepatocellular carcinoma: analysis of mutation and expression of beta-catenin, T‐cell factor‐4 and glycogen synthase kinase 3‐beta genes. J Gastroenterol Hepatol 18(3):280–287
Shi J, Keller J, Zhang J, Keller E (2014) A review on the diagnosis and treatment of hepatocellular carcinoma with a focus on the role of wnts and the dickkopf family of wnt inhibitors. J Hepato Carcinoma 1:1–7
Alao JP (2007) The regulation of cyclin D1 degradation: roles in cancer development and the potential for therapeutic invention. Mol Cancer 6(1):24
Xia H, Ng SS, Jiang S, Cheung WK, Sze J, Bian X-W, Kung H-f, Lin MC (2010) miR-200a-mediated downregulation of ZEB2 and CTNNB1 differentially inhibits nasopharyngeal carcinoma cell growth, migration and invasion. Biochem Biophys Res Commun 391(1):535–541
Abedi N, Mohammadi-Yeganeh S, Koochaki A, Karami F, Paryan M (2015) miR-141 as potential suppressor of β-catenin in breast cancer. Tumor Biol 36:1–7
Le Quesne J, Caldas C (2010) Micro-RNAs and breast cancer. Mol Oncol 4(3):230–241
Schetter AJ, Leung SY, Sohn JJ, Zanetti KA, Bowman ED, Yanaihara N, Yuen ST, Chan TL, Kwong DL, Au GK (2008) MicroRNA expression profiles associated with prognosis and therapeutic outcome in colon adenocarcinoma. JAMA 299(4):425–436
Wang B, Xi Y (2013) Challenges for microRNA microarray data analysis. Microarrays 2(2):34–50
Wahid F, Khan T, Kim YY (2014) MicroRNA and diseases: therapeutic potential as new generation of drugs. Biochimie 104:12–26
Gong J, He X-X, Tian D-A (2015) Emerging role of microRNA in hepatocellular carcinoma (review). Oncol Lett 9(3):1027–1033
Murakami Y, Yasuda T, Saigo K, Urashima T, Toyoda H, Okanoue T, Shimotohno K (2006) Comprehensive analysis of microRNA expression patterns in hepatocellular carcinoma and non-tumorous tissues. Oncogene 25(17):2537–2545
Bader AG (2012) miR-34–a microRNA replacement therapy is headed to the clinic. Front Genet 3:120
Callegari E, Elamin BK, Giannone F, Milazzo M, Altavilla G, Fornari F, Giacomelli L, D’Abundo L, Ferracin M, Bassi C (2012) Liver tumorigenicity promoted by microRNA-221 in a mouse transgenic model. Hepatology 56(3):1025–1033
Zhang J, Jiao J, Cermelli S, Muir K, Jung KH, Zou R, Rashid A, Gagea M, Zabludoff S, Kalluri R (2015) miR-21 inhibition reduces liver fibrosis and prevents tumor development by inducing apoptosis of CD24 + progenitor cells. Cancer Res 75(9):1859–1867
Yang N, Ekanem NR, Sakyi CA, Ray SD (2015) Hepatocellular carcinoma and microRNA: new perspectives on therapeutics and diagnostics. Adv Drug Deliv Rev 81:62–74
Ritchie W, Rasko JE, Flamant S (2013) MicroRNA target prediction and validation. In: Schmitz U, Wolkenhauer O, Vera J (eds) MicroRNA cancer regulation. Springer, Dordrecht, pp 39–53
Heyn H, Schreek S, Buurman R, Focken T, Schlegelberger B, Beger C (2012) MicroRNA miR-548d is a superior regulator in pancreatic cancer. Pancreas 41(2):218–221
Li Y, Zhang Z (2015) Computational biology in microRNA. Wiley, Hoboken
Cho WC (2010) MicroRNAs in cancer—from research to therapy. Biochim Biophys Acta (BBA) 1805(2):209–217
Tsao CM, Yan MD, Shih YL, Yu PN, Kuo CC, Lin WC, Li HJ, Lin YW (2012) SOX1 functions as a tumor suppressor by antagonizing the WNT/β-catenin signaling pathway in hepatocellular carcinoma. Hepatology 56(6):2277–2287
Gyöngyösi B, Végh É, Járay B, Székely E, Fassan M, Bodoky G, Schaff Z, Kiss A (2014) Pretreatment MicroRNA level and outcome in sorafenib-treated hepatocellular carcinoma. J Histochem Cytochem 62(8):547–555
Zhang Z, Yin J, Yang J, Shen W, Zhang C, Mou W, Luo J, Yan H, Sun P, Luo Y (2016) miR-885-5p suppresses hepatocellular carcinoma metastasis and inhibits Wnt/β-catenin signaling pathway. Oncotarget 7(46):75038
Tao J, Zhang R, Singh S, Poddar M, Xu E, Oertel M, Chen X, Ganesh S, Abrams M, Monga SP (2017) Targeting β-catenin in hepatocellular cancers induced by coexpression of mutant β‐catenin and K‐Ras in mice. Hepatology 65(5):1581–1599
Zhang K-C, Xi H-Q, Cui J-X, Shen W-S, Li J-Y, Wei B, Chen L (2015) Hemolysis-free plasma miR-214 as novel biomarker of gastric cancer and is correlated with distant metastasis. Am J Cancer Res 5(2):821
Acknowledgements
This study was funded by Pasteur Institute of Iran, Tehran, Iran. The authors should thank Stem Cell technology Research Center, Tehran, Iran, and also Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences for providing technical supports.
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare no conflict of interest.
Informed consent
It is not applicable in this study because the research was completely based on in vitro investigation and no human samples have been used in this project.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Karimkhanloo, H., Mohammadi-Yeganeh, S., Hadavi, R. et al. Potential role of miR-214 in β-catenin gene expression within hepatocellular carcinoma. Mol Biol Rep 47, 7429–7437 (2020). https://doi.org/10.1007/s11033-020-05798-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-020-05798-5