Abstract
Emerging evidence has shown the association of aberrantly expressed microRNAs (miRNAs) with tumor development and progression. However, little is known about the potential role of miRNAs in gastric carcinogenesis. Here, we performed miRNA microarray to screen miRNAs differentially expressed in the paired gastric cancer and their adjacent nontumor tissues and found that miR-375 was greatly downregulated in gastric cancer tissues. Quantitative real-time PCR analysis verified that miR-375 expression was significantly decreased in more than 90% of primary gastric cancers compared with their nontumor counterparts from patients undergoing gastric resection. Overexpression of miR-375 significantly inhibited gastric cancer cell proliferation in vitro and in vivo. Forced expression of miR-375 in gastric cancer cells significantly reduced the protein level of Janus kinase 2 (JAK2) and repressed the activity of a luciferase reporter carrying the 3′-untranslated region of JAK2, which was abolished by mutation of the predicted miR-375-binding site, indicating that JAK2 may be a miR-375 target gene. Either inhibition of JAK2 activity by AG490 or silencing of JAK2 by RNAi suppressed gastric cancer cell proliferation resembling that of miR-375 overexpression. Moreover, ectopic expression of JAK2 can partially reverse the inhibition of cell proliferation caused by miR-375. Finally, we found a significant inverse correlation between miR-375 expression and JAK2 protein level in gastric cancer. Thus, these data suggest that miR-375 may function as a tumor suppressor to regulate gastric cancer cell proliferation potentially by targeting the JAK2 oncogene, implicating a role of miR-375 in the pathogenesis of gastric cancer.
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Abbreviations
- miRNA:
-
(microRNA)
- JAK2:
-
(Janus kinase 2)
- 3′-UTR:
-
(3′-untranslated region)
- qRT-PCR:
-
(quantitative real-time RT-PCR)
- STAT:
-
(signal transducer and activator of transcription)
References
Ambros V . The functions of animal microRNAs. Nature 2004; 431:350–355.
Bartel DP . MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 2004; 116:281–297.
Pasquinelli AE, Hunter S, Bracht J . MicroRNAs: a developing story. Curr Opin Genet Dev 2005; 15:200–205.
Ahmed FE . Role of miRNA in carcinogenesis and biomarker selection: a methodological view. Expert Rev Mol Diagn 2007; 7:569–603.
Calin GA, Sevignani C, Dumitru CD, et al. Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers. Proc Natl Acad Sci USA 2004; 101:2999–3004.
Croce CM . Causes and consequences of microRNA dysregulation in cancer. Nat Rev Genet 2009; 10:704–714.
Parkin DM, Bray F, Ferlay J, Pisani P . Global cancer statistics, 2002. CA Cancer J Clin 2005; 55:74–108.
Hohenberger P, Gretschel S . Gastric cancer. Lancet 2003; 362:305–315.
Yasui W, Yokozaki H, Fujimoto J, et al. Genetic and epigenetic alterations in multistep carcinogenesis of the stomach. J Gastroenterol 2000; 35 Suppl 12:111–115.
Stock M, Otto F . Gene deregulation in gastric cancer. Gene 2005; 360:1–19.
Du Y, Xu Y, Ding L, et al. Down-regulation of miR-141 in gastric cancer and its involvement in cell growth. J Gastroenterol 2009; 44:556–561.
Takagi T, Iio A, Nakagawa Y, et al. Decreased expression of microRNA-143 and -145 in human gastric cancers. Oncology 2009; 77:12–21.
Luo H, Zhang H, Zhang Z, et al. Down-regulated miR-9 and miR-433 in human gastric carcinoma. J Exp Clin Cancer Res 2009; 28:82.
Gao C, Zhang Z, Liu W, et al. Reduced microRNA-218 expression is associated with high nuclear factor kappa B activation in gastric cancer. Cancer; 116:41–49.
Chan SH, Wu CW, Li AF, Chi CW, Lin WC . miR-21 microRNA expression in human gastric carcinomas and its clinical association. Anticancer Res 2008; 28:907–911.
Wu Q, Jin H, Yang Z, et al. MiR-150 promotes gastric cancer proliferation by negatively regulating the pro-apoptotic gene EGR2. Biochem Biophys Res Commun 2010; 392:340–345.
Xiao B, Guo J, Miao Y, et al. Detection of miR-106a in gastric carcinoma and its clinical significance. Clin Chim Acta 2009; 400:97–102.
Zhang Z, Li Z, Gao C, et al. miR-21 plays a pivotal role in gastric cancer pathogenesis and progression. Lab Invest 2008; 88:1358–1366.
Petrocca F, Visone R, Onelli MR, et al. E2F1-regulated microRNAs impair TGFbeta-dependent cell-cycle arrest and apoptosis in gastric cancer. Cancer Cell 2008; 13:272–286.
Kim YJ, Hwang SJ, Bae YC, Jung JS . MiR-21 regulates adipogenic differentiation through the modulation of TGF-beta signaling in mesenchymal stem cells derived from human adipose tissue. Stem Cells 2009; 27:3093–3102.
Meydan N, Grunberger T, Dadi H, et al. Inhibition of acute lymphoblastic leukaemia by a Jak-2 inhibitor. Nature 1996; 379:645–648.
Poy MN, Eliasson L, Krutzfeldt J, et al. A pancreatic islet-specific microRNA regulates insulin secretion. Nature 2004; 432:226–230.
El Ouaamari A, Baroukh N, Martens GA, et al. miR-375 targets 3′-phosphoinositide-dependent protein kinase-1 and regulates glucose-induced biological responses in pancreatic beta-cells. Diabetes 2008; 57:2708–2717.
Poy MN, Hausser J, Trajkovski M, et al. miR-375 maintains normal pancreatic alpha- and beta-cell mass. Proc Natl Acad Sci USA 2009; 106:5813–5818.
Kloosterman WP, Lagendijk AK, Ketting RF, Moulton JD, Plasterk RH . Targeted inhibition of miRNA maturation with morpholinos reveals a role for miR-375 in pancreatic islet development. PLoS Biol 2007; 5:e203.
Ladeiro Y, Couchy G, Balabaud C, et al. MicroRNA profiling in hepatocellular tumors is associated with clinical features and oncogene/tumor suppressor gene mutations. Hepatology 2008; 47:1955–1963.
Avissar M, Christensen BC, Kelsey KT, Marsit CJ . MicroRNA expression ratio is predictive of head and neck squamous cell carcinoma. Clin Cancer Res 2009; 15:2850–2855.
Bloomston M, Frankel WL, Petrocca F, et al. MicroRNA expression patterns to differentiate pancreatic adenocarcinoma from normal pancreas and chronic pancreatitis. JAMA 2007; 297:1901–1908.
Mathe EA, Nguyen GH, Bowman ED, et al. MicroRNA expression in squamous cell carcinoma and adenocarcinoma of the esophagus: associations with survival. Clin Cancer Res 2009; 15:6192–6200.
Constantinescu SN, Girardot M, Pecquet C . Mining for JAK-STAT mutations in cancer. Trends Biochem Sci 2008; 33:122–131.
Levine RL, Pardanani A, Tefferi A, Gilliland DG . Role of JAK2 in the pathogenesis and therapy of myeloproliferative disorders. Nat Rev Cancer 2007; 7:673–683.
Levine RL, Wadleigh M, Cools J, et al. Activating mutation in the tyrosine kinase JAK2 in polycythemia vera, essential thrombocythemia, and myeloid metaplasia with myelofibrosis. Cancer Cell 2005; 7:387–397.
James C, Ugo V, Le Couedic JP, et al. A unique clonal JAK2 mutation leading to constitutive signalling causes polycythaemia vera. Nature 2005; 434:1144–1148.
Baxter EJ, Scott LM, Campbell PJ, et al. Acquired mutation of the tyrosine kinase JAK2 in human myeloproliferative disorders. Lancet 2005; 365:1054–1061.
Kralovics R, Passamonti F, Buser AS, et al. A gain-of-function mutation of JAK2 in myeloproliferative disorders. N Engl J Med 2005; 352:1779–1790.
Lee JW, Soung YH, Kim SY, et al. Absence of JAK2 V617F mutation in gastric cancers. Acta Oncol 2006; 45:222–223.
Kumar C, Purandare AV, Lee FY, Lorenzi MV . Kinase drug discovery approaches in chronic myeloproliferative disorders. Oncogene 2009; 28:2305–2313.
Orom UA, Kauppinen S, Lund AH . LNA-modified oligonucleotides mediate specific inhibition of microRNA function. Gene 2006; 372:137–141.
Zhou T, Zimmerman W, Liu X, Erikson RL . A mammalian NudC-like protein essential for dynein stability and cell viability. Proc Natl Acad Sci USA 2006; 103:9039–9044.
Cai Y, Yang Y, Shen M, Zhou T . Inhibition of cytokinesis by overexpression of NudCL that is localized to the centrosome and midbody. Cell Res 2009; 19:1305–1308.
Acknowledgements
This work was supported by the National Natural Scientific Foundation of China (30901714, 30671070 and 30771107), the Ministry of Science and Technology of China (2007CB914500), the Ministry of Education of China (NCET-06-0530), the Ministry of Health of China (WKJ2006-2-014), the Postdoctoral Science Foundation of China (20070421179), the Department of Science and Technology of Zhejiang Province (2009F80032), and the Natural Scientific Foundation of Zhejiang Province, China (R205291, Y206103 and 2007R10G2010103).
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( Supplementary information is linked to the online version of the paper on Cell Research website.)
Supplementary information
Supplementary information, Table S1
Clinical and pathological parameters of included patients from gastric resection (PDF 11 kb)
Supplementary information, Table S2
Clinical characteristics of included patients from gastroscopy (PDF 9 kb)
Supplementary information, Figure S1
MiR-375 expression is significantly downregulated in five gastric cancer cell lines compared to gastric normal tissues. (PDF 90 kb)
Supplementary information, Figure S2
Relative expression of miR-375 in gastric cancer cells transfected with miR-375 precursor. (PDF 73 kb)
Supplementary information, Figure S3
Effects of miR-375 on the proliferation of gastric cancer cells. (PDF 77 kb)
Supplementary information, Figure S4
The expression of JAK2 mRNA in gastric cancer cells with transfection of miR-375. (PDF 71 kb)
Supplementary information, Figure S5
The protein levels of JAK2 in AGS and MGC-803 cells treated with vector-based RNAi and JAK2 overexpression. (PDF 75 kb)
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Ding, L., Xu, Y., Zhang, W. et al. MiR-375 frequently downregulated in gastric cancer inhibits cell proliferation by targeting JAK2. Cell Res 20, 784–793 (2010). https://doi.org/10.1038/cr.2010.79
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DOI: https://doi.org/10.1038/cr.2010.79
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