Differentially Expressed miRNAs in Hepatocellular Carcinoma Target Genes in the Genetic Information Processing and Metabolism Pathways

To date, studies of the roles of microRNAs (miRNAs) in hepatocellular carcinoma (HCC) have either focused on specific individual miRNAs and a small number of suspected targets or simply reported a list of differentially expressed miRNAs based on expression profiling. Here, we seek a more in-depth understanding of the roles of miRNAs and their targets in HCC by integrating the miRNA and messenger RNA (mRNA) expression profiles of tumorous and adjacent non-tumorous liver tissues of 100 HCC patients. We assessed the levels of 829 mature miRNAs, of which 32 were significantly differentially expressed. Statistical analysis indicates that six of these miRNAs regulate a significant proportion of their in silico predicted target mRNAs. Three of these miRNAs (miR-26a, miR-122, and miR-130a) were down-regulated in HCC, and their up-regulated gene targets are primarily associated with aberrant cell proliferation that involves DNA replication, transcription and nucleotide metabolism. The other three miRNAs (miR-21, miR-93, and miR-221) were up-regulated in HCC, and their down-regulated gene targets are primarily involved in metabolism and immune system processes. We further found evidence for a coordinated miRNA-induced regulation of important cellular processes, a finding to be considered when designing therapeutic applications based on miRNAs.

[1]  C. Croce,et al.  MiR-122/cyclin G1 interaction modulates p53 activity and affects doxorubicin sensitivity of human hepatocarcinoma cells. , 2009, Cancer research.

[2]  Chang-Zheng Chen,et al.  MicroRNAs as oncogenes and tumor suppressors. , 2005, The New England journal of medicine.

[3]  Kuo-Bin Li,et al.  Profiling MicroRNA Expression in Hepatocellular Carcinoma Reveals MicroRNA-224 Up-regulation and Apoptosis Inhibitor-5 as a MicroRNA-224-specific Target* , 2008, Journal of Biological Chemistry.

[4]  P. Chow,et al.  MicroRNA-224 Targets SMAD Family Member 4 to Promote Cell Proliferation and Negatively Influence Patient Survival , 2013, PloS one.

[5]  Xiuping Liu,et al.  Regulation of the Extrinsic Apoptotic Pathway by MicroRNA-21 in Alcoholic Liver Injury* , 2014, The Journal of Biological Chemistry.

[6]  P. Anthony Hepatocellular carcinoma: an overview , 2001, Histopathology.

[7]  Z. Duan,et al.  Interleukin-6 signaling pathway in targeted therapy for cancer. , 2012, Cancer treatment reviews.

[8]  Doron Betel,et al.  The microRNA.org resource: targets and expression , 2007, Nucleic Acids Res..

[9]  Gordon K Smyth,et al.  Statistical Applications in Genetics and Molecular Biology Linear Models and Empirical Bayes Methods for Assessing Differential Expression in Microarray Experiments , 2011 .

[10]  C. Croce,et al.  miR-130a targets MET and induces TRAIL-sensitivity in NSCLC by downregulating miR-221 and 222 , 2012, Oncogene.

[11]  Ming-Sound Tsao,et al.  An overview of the c-MET signaling pathway , 2011, Therapeutic advances in medical oncology.

[12]  A. Krook,et al.  Role of interleukin‐6 signalling in glucose and lipid metabolism , 2007, Acta physiologica.

[13]  J. Ranish,et al.  Isolation and Characterization of Proteins Associated with Histone H3 Tails in Vivo* , 2007, Journal of Biological Chemistry.

[14]  Zhaolei Zhang,et al.  Potential microRNA-mediated oncogenic intercellular communication revealed by pan-cancer analysis , 2014, Scientific Reports.

[15]  Y. Benjamini,et al.  Controlling the false discovery rate: a practical and powerful approach to multiple testing , 1995 .

[16]  J. Llovet,et al.  Molecular classification and novel targets in hepatocellular carcinoma: recent advancements. , 2010, Seminars in liver disease.

[17]  J. Neuzil,et al.  MicroRNA regulation of cancer metabolism: role in tumour suppression. , 2014, Mitochondrion.

[18]  Xiaowei Wang,et al.  Sequence analysis Prediction of both conserved and nonconserved microRNA targets in animals , 2007 .

[19]  P. Berk Seminars in Liver Disease , 2014 .

[20]  Jianxing He,et al.  MicroRNA-26a/b Regulate DNA Replication Licensing, Tumorigenesis, and Prognosis by Targeting CDC6 in Lung Cancer , 2014, Molecular Cancer Research.

[21]  A. Jemal,et al.  Global cancer statistics , 2011, CA: a cancer journal for clinicians.

[22]  K. Ghoshal,et al.  Downregulation of miR‐122 in the rodent and human hepatocellular carcinomas , 2006, Journal of cellular biochemistry.

[23]  S. Lowe,et al.  Rb-Mediated Heterochromatin Formation and Silencing of E2F Target Genes during Cellular Senescence , 2003, Cell.

[24]  C. Croce,et al.  MiR-221 controls CDKN1C/p57 and CDKN1B/p27 expression in human hepatocellular carcinoma , 2008, Oncogene.

[25]  M. Oren,et al.  The Lats2 tumor suppressor augments p53-mediated apoptosis by promoting the nuclear proapoptotic function of ASPP1. , 2010, Genes & development.

[26]  Mark Graham,et al.  miR-122 regulation of lipid metabolism revealed by in vivo antisense targeting. , 2006, Cell metabolism.

[27]  M. Caligiuri,et al.  Essential metabolic, anti-inflammatory, and anti-tumorigenic functions of miR-122 in liver. , 2012, The Journal of clinical investigation.

[28]  Gordon K. Smyth,et al.  A comparison of background correction methods for two-colour microarrays , 2007, Bioinform..

[29]  J. Scheller,et al.  Therapeutic strategies for the clinical blockade of IL-6/gp130 signaling. , 2011, The Journal of clinical investigation.

[30]  Rounak Nassirpour,et al.  miR-122 Regulates Tumorigenesis in Hepatocellular Carcinoma by Targeting AKT3 , 2013, PloS one.

[31]  Zhao-You Tang,et al.  MicroRNA-26a Inhibits Angiogenesis by Down-Regulating VEGFA through the PIK3C2α/Akt/HIF-1α Pathway in Hepatocellular Carcinoma , 2013, PloS one.

[32]  Joshua M. Stuart,et al.  The Cancer Genome Atlas Pan-Cancer analysis project , 2013, Nature Genetics.

[33]  Yu Wang,et al.  Role of miR-224 in hepatocellular carcinoma: a tool for possible therapeutic intervention? , 2011, Epigenomics.

[34]  C. Croce,et al.  Cyclin G1 is a target of miR-122a, a microRNA frequently down-regulated in human hepatocellular carcinoma. , 2007, Cancer research.

[35]  Shireen A. Sarraf,et al.  Methyl-CpG binding protein MBD1 couples histone H3 methylation at lysine 9 by SETDB1 to DNA replication and chromatin assembly. , 2004, Molecular cell.

[36]  L. Baier,et al.  Human adipose microRNA-221 is upregulated in obesity and affects fat metabolism downstream of leptin and TNF-α , 2013, Diabetologia.

[37]  Kristian Helin,et al.  The Polycomb group proteins bind throughout the INK4A-ARF locus and are disassociated in senescent cells. , 2007, Genes & development.

[38]  J. Nielsen,et al.  Expression and Localization of microRNAs in Perinatal Rat Pancreas: Role of miR-21 in Regulation of Cholesterol Metabolism , 2011, PloS one.

[39]  D. Iliopoulos,et al.  miRNAs link metabolic reprogramming to oncogenesis , 2013, Trends in Endocrinology & Metabolism.

[40]  Li Lin,et al.  Identification of miRNomes in human liver and hepatocellular carcinoma reveals miR-199a/b-3p as therapeutic target for hepatocellular carcinoma. , 2011, Cancer cell.

[41]  Pablo Tamayo,et al.  Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[42]  Xia Zhao,et al.  Roles of microRNA on cancer cell metabolism , 2012, Journal of Translational Medicine.

[43]  D. Cui,et al.  Tumor-specific expression of microRNA-26a suppresses human hepatocellular carcinoma growth via cyclin-dependent and -independent pathways. , 2011, Molecular therapy : the journal of the American Society of Gene Therapy.

[44]  D. Stepp,et al.  miRNA-93 Inhibits GLUT4 and Is Overexpressed in Adipose Tissue of Polycystic Ovary Syndrome Patients and Women With Insulin Resistance , 2013, Diabetes.

[45]  Mary Goldman,et al.  The UCSC Genome Browser database: extensions and updates 2011 , 2011, Nucleic Acids Res..

[46]  W. Sellers,et al.  Drug discovery approaches targeting the PI3K/Akt pathway in cancer , 2008, Oncogene.

[47]  C. Sardet,et al.  Coactivator-associated arginine methyltransferase 1 (CARM1) is a positive regulator of the Cyclin E1 gene , 2006, Proceedings of the National Academy of Sciences.

[48]  Michael W Pfaffl,et al.  RNA integrity and the effect on the real-time qRT-PCR performance. , 2006, Molecular aspects of medicine.

[49]  Y. Homma,et al.  TACC2 is an androgen-responsive cell cycle regulator promoting androgen-mediated and castration-resistant growth of prostate cancer. , 2012, Molecular endocrinology.

[50]  Mary Goldman,et al.  The UCSC Genome Browser database: extensions and updates 2013 , 2012, Nucleic Acids Res..

[51]  K. Ghoshal,et al.  MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer. , 2007, Gastroenterology.

[52]  Terry Speed,et al.  Normalization of cDNA microarray data. , 2003, Methods.

[53]  M. Manns,et al.  MicroRNA‐221 regulates FAS‐induced fulminant liver failure , 2011, Hepatology.

[54]  Stephanie Roessler,et al.  MicroRNA expression, survival, and response to interferon in liver cancer. , 2009, The New England journal of medicine.

[55]  Aaron N. Chang,et al.  MicroRNA-21 Promotes Fibrosis of the Kidney by Silencing Metabolic Pathways , 2012, Science Translational Medicine.

[56]  L. Holmberg,et al.  Integrated genomic analysis of triple-negative breast cancers reveals novel microRNAs associated with clinical and molecular phenotypes and sheds light on the pathways they control , 2013, BMC Genomics.

[57]  Stijn van Dongen,et al.  miRBase: microRNA sequences, targets and gene nomenclature , 2005, Nucleic Acids Res..

[58]  Q. Ye,et al.  MicroRNA‐26a suppresses tumor growth and metastasis of human hepatocellular carcinoma by targeting interleukin‐6‐Stat3 pathway , 2013, Hepatology.

[59]  K. Yokomori,et al.  A Human Condensin Complex Containing hCAP-C–hCAP-E and CNAP1, a Homolog of Xenopus XCAP-D2, Colocalizes with Phosphorylated Histone H3 during the Early Stage of Mitotic Chromosome Condensation , 2000, Molecular and Cellular Biology.

[60]  Xiaowei Wang miRDB: a microRNA target prediction and functional annotation database with a wiki interface. , 2008, RNA.

[61]  D. Waugh,et al.  Rationale and Means to Target Pro-Inflammatory Interleukin-8 (CXCL8) Signaling in Cancer , 2013, Pharmaceuticals.

[62]  S. Lowe,et al.  579 MIR-221 OVEREXPRESSION CONTRIBUTES TO LIVER TUMORIGENESIS , 2010 .

[63]  Jian-Rong Yang,et al.  MicroRNA-26a/b and their host genes cooperate to inhibit the G1/S transition by activating the pRb protein , 2011, Nucleic acids research.

[64]  Helga Thorvaldsdóttir,et al.  Molecular signatures database (MSigDB) 3.0 , 2011, Bioinform..

[65]  C. Lee,et al.  MicroRNA and cancer – focus on apoptosis , 2008, Journal of cellular and molecular medicine.

[66]  Cheng Li,et al.  Adjusting batch effects in microarray expression data using empirical Bayes methods. , 2007, Biostatistics.

[67]  Anton J. Enright,et al.  Human MicroRNA Targets , 2004, PLoS biology.

[68]  P. Dennis,et al.  PTEN loss in the continuum of common cancers, rare syndromes and mouse models , 2011, Nature Reviews Cancer.

[69]  S. Lowe,et al.  miR-221 overexpression contributes to liver tumorigenesis , 2009, Proceedings of the National Academy of Sciences.

[70]  Angela M. Liu,et al.  microRNA-122 as a regulator of mitochondrial metabolic gene network in hepatocellular carcinoma , 2010, Molecular systems biology.