Systematic analysis of the molecular mechanism of microRNA‑124 in hepatoblastoma cells

Wang,Guiming; Liu,Hong; Wei,Zhigang; Jia,Hongyan; Liu,Yu; Liu,Jiansheng

doi:10.3892/ol.2017.7131

Systematic analysis of the molecular mechanism of microRNA‑124 in hepatoblastoma cells

Authors:
- Guiming Wang
- Hong Liu
- Zhigang Wei
- Hongyan Jia
- Yu Liu
- Jiansheng Liu
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Affiliations: Department of Surgery, The First Hospital of Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China
Published online on: October 3, 2017 https://doi.org/10.3892/ol.2017.7131
Pages: 7161-7170
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study aimed to identify the molecular mechanisms of microRNA‑124 (miRNA‑124/miR‑124) in hepatoblastoma. The GSE6207 microarray dataset, obtained from the Gene Expression Omnibus database, included samples extracted from HepG2 cells transfected with miR‑124 duplex (the experimental group) or negative control (the control group) at 4, 8, 16, 24, 32, 72 and 120 h after transfection. Differentially expressed genes (DEGs) were screened between the two groups. miR‑124 activity was inferred based on the expression of its target genes. The mRNAs targeted by miR‑124 were predicted and a miR‑124‑target mRNA network was constructed. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed for the target genes. The number of DEGs was highest at 72 h. The experimental group had higher miR‑124 activity than that of the control group at 4, 8, 16, 24 and 120 h. Small GTPase‑mediated signal transduction and Ras protein signal transduction were significant GO terms enriched with syndecan binding protein (SDCBP), Ras homolog family member G (RHOG) and Rho‑GDP dissociation inhibitor‑α (ARHGDIA). Regulation of actin cytoskeleton, D‑glutamine and D‑glutamate metabolism, and axon guidance were significant pathways. Axon guidance pathway was associated with neuropilin (NRP1), MET proto‑oncogene, receptor tyrosine kinase (MET) and semaphorin 7A, GPI membrane anchor (SEMA7A). Small GTPase‑mediated signal transduction, Ras protein signal transduction, regulation of actin cytoskeleton pathway, D‑glutamine and D‑glutamate metabolism pathway, axon guidance pathway, SDCBP, RHOG, ARHGDIA, NRP1, SEMA7A, and MET may be implicated in the underlying mechanisms of miR‑124 overexpression in hepatoblastoma.

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1	Stewart BW and Wild CP: World cancer report 2014. World Health Organization. 2014.
2	Ringe B, Pichlmayr R, Wittekind C and Tusch G: Surgical treatment of hepatocellular carcinoma: Experience with liver resection and transplantation in 198 patients. World J Surg. 15:270–285. 1991. View Article : Google Scholar : PubMed/NCBI
3	Maluccio M and Covey A: Recent progress in understanding, diagnosing, and treating hepatocellular carcinoma. CA Cancer J Clin. 62:394–399. 2012. View Article : Google Scholar : PubMed/NCBI
4	Bruix J and Sherman M: American Association for the Study of Liver Diseases: Management of hepatocellular carcinoma: An update. Hepatology. 53:1020–1022. 2011. View Article : Google Scholar : PubMed/NCBI
5	Meyers RL, Tiao G, de Ville de Goyet J, Superina R and Aronson DC: Hepatoblastoma state of the art: Pre-treatment extent of disease, surgical resection guidelines and the role of liver transplantation. Curr Opin Pediatr. 26:29–36. 2014. View Article : Google Scholar : PubMed/NCBI
6	Osada H and Takahashi T: MicroRNAs in biological processes and carcinogenesis. Carcinogenesis. 28:2–12. 2007. View Article : Google Scholar : PubMed/NCBI
7	Shi XB, Xue L, Ma AH, Tepper CG, Gandour-Edwards R, Kung HJ and deVere White RW: Tumor suppressive miR-124 targets androgen receptor and inhibits proliferation of prostate cancer cells. Oncogene. 32:4130–4138. 2013. View Article : Google Scholar : PubMed/NCBI
8	Zheng H, Song F, Zhang L, Yang D, Ji P, Wang Y, Almeida M, Calin GA, Hao X, Wei Q, et al: Genetic variants at the miR-124 binding site on the cytoskeleton-organizing IQGAP1 gene confer differential predisposition to breast cancer. Int J Oncol. 38:1153–1161. 2011.PubMed/NCBI
9	Zheng F, Liao YJ, Cai MY, Liu YH, Liu TH, Chen SP, Bian XW, Guan XY, Lin MC, Zeng YX, et al: The putative tumour suppressor microRNA-124 modulates hepatocellular carcinoma cell aggressiveness by repressing ROCK2 and EZH2. Gut. 61:278–289. 2012. View Article : Google Scholar : PubMed/NCBI
10	Zeng B, Li Z, Chen R, Guo N, Zhou J, Zhou Q, Lin Q, Cheng D, Liao Q, Zheng L and Gong Y: Epigenetic regulation of miR-124 by hepatitis C virus core protein promotes migration and invasion of intrahepatic cholangiocarcinoma cells by targeting SMYD3. FEBS Lett. 586:3271–3278. 2012. View Article : Google Scholar : PubMed/NCBI
11	Krützfeldt J, Rajewsky N, Braich R, Rajeev KG, Tuschl T, Manoharan M and Stoffel M: Silencing of microRNAs in vivo with ‘antagomirs’. Nature. 438:685–689. 2005. View Article : Google Scholar : PubMed/NCBI
12	Cheng C and Li LM: Inferring microRNA activities by combining gene expression with microRNA target prediction. PLoS One. 3:e19892008. View Article : Google Scholar : PubMed/NCBI
13	Lim LP, Lau NC, Garrett-Engele P, Grimson A, Schelter JM, Castle J, Bartel DP, Linsley PS and Johnson JM: Microarray analysis shows that some microRNAs downregulate large numbers of target mRNAs. Nature. 433:769–773. 2005. View Article : Google Scholar : PubMed/NCBI
14	Wang X and Wang X: Systematic identification of microRNA functions by combining target prediction and expression profiling. Nucleic Acids Res. 34:1646–1652. 2006. View Article : Google Scholar : PubMed/NCBI
15	Irizarry RA, Hobbs B, Collin F, Beazer-Barclay YD, Antonellis KJ, Scherf U and Speed TP: Exploration, normalization and summaries of high density oligonucleotide array probe level data. Biostatistics. 4:249–264. 2003. View Article : Google Scholar : PubMed/NCBI
16	Bernhardt V, Hotchkiss MT, Garcia-Reyero N, Escalon BL, Denslow N and Davenport PW: Tracheal occlusion conditioning in conscious rats modulates gene expression profile of medial thalamus. Front Physiol. 2:242011. View Article : Google Scholar : PubMed/NCBI
17	Shannon W, Culverhouse R and Duncan J: Analyzing microarray data using cluster analysis. Pharmacogenomics. 4:41–52. 2003. View Article : Google Scholar : PubMed/NCBI
18	Huang da W, Sherman BT and Lempicki RA: Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc. 4:44–57. 2009. View Article : Google Scholar : PubMed/NCBI
19	Lewis BP, Shih IH, Jones-Rhoades MW, Bartel DP and Burge CB: Prediction of mammalian microRNA targets. Cell. 115:787–798. 2003. View Article : Google Scholar : PubMed/NCBI
20	Kertesz M, Iovino N, Unnerstall U, Gaul U and Segal E: The role of site accessibility in microRNA target recognition. Nat Genet. 39:1278–1284. 2007. View Article : Google Scholar : PubMed/NCBI
21	Lall S, Grün D, Krek A, Chen K, Wang YL, Dewey CN, Sood P, Colombo T, Bray N, Macmenamin P, et al: A genome-wide map of conserved microRNA targets in C. elegans. Curr Biol. 16:460–471. 2006. View Article : Google Scholar : PubMed/NCBI
22	Wang X: miRDB: A microRNA target prediction and functional annotation database with a wiki interface. RNA. 14:1012–1017. 2008. View Article : Google Scholar : PubMed/NCBI
23	Betel D, Wilson M, Gabow A, Marks DS and Sander C: The microRNA.org resource: targets and expression. Nucleic Acids Res. 36:D149–D153. 2008. View Article : Google Scholar : PubMed/NCBI
24	Harris MA, Clark J, Ireland A, Lomax J, Ashburner M, Foulger R, Eilbeck K, Lewis S, Marshall B, Mungall C, et al: The gene ontology (GO) database and informatics resource. Nucleic Acids Res. 32:(Database Issue). D258–D261. 2004. View Article : Google Scholar : PubMed/NCBI
25	Weis SM and Cheresh DA: Tumor angiogenesis: Molecular pathways and therapeutic targets. Nat Med. 17:1359–1370. 2011. View Article : Google Scholar : PubMed/NCBI
26	Pillai RS: MicroRNA function: Multiple mechanisms for a tiny RNA? RNA. 11:1753–1761. 2005. View Article : Google Scholar : PubMed/NCBI
27	Hobert O: Common logic of transcription factor and microRNA action. Trends Biochem Sci. 29:462–468. 2004. View Article : Google Scholar : PubMed/NCBI
28	Etienne-Manneville S and Hall A: Rho GTPases in cell biology. Nature. 420:629–635. 2002. View Article : Google Scholar : PubMed/NCBI
29	Jaffe AB and Hall A: Rho GTPases: Biochemistry and biology. Annu Rev Cell Dev Biol. 21:247–269. 2005. View Article : Google Scholar : PubMed/NCBI
30	Yoshimura T, Arimura N, Kawano Y, Kawabata S, Wang S and Kaibuchi K: Ras regulates neuronal polarity via the PI3-kinase/Akt/GSK-3beta/CRMP-2 pathway. Biochem Biophys Res Commun. 340:62–68. 2006. View Article : Google Scholar : PubMed/NCBI
31	Chang L and Karin M: Mammalian MAP kinase signalling cascades. Nature. 410:37–40. 2001. View Article : Google Scholar : PubMed/NCBI
32	Wong CM, Yam JW, Ching YP, Yau TO, Leung TH, Jin DY and Ng IO: Rho GTPase-activating protein deleted in liver cancer suppresses cell proliferation and invasion in hepatocellular carcinoma. Cancer Res. 65:8861–8868. 2005. View Article : Google Scholar : PubMed/NCBI
33	Calvisi DF, Ladu S, Conner EA, Seo D, Hsieh JT, Factor VM and Thorgeirsson SS: Inactivation of Ras GTPase-activating proteins promotes unrestrained activity of wild-type Ras in human liver cancer. J Hepatol. 54:311–319. 2011. View Article : Google Scholar : PubMed/NCBI
34	Beekman JM and Coffer PJ: The ins and outs of syntenin, a multifunctional intracellular adaptor protein. J Cell Sci. 121:1349–1355. 2008. View Article : Google Scholar : PubMed/NCBI
35	Lee H, Kim Y, Choi Y, Choi S, Hong E and Oh ES: Syndecan-2 cytoplasmic domain regulates colon cancer cell migration via interaction with syntenin-1. Biochem Biophys Res Commun. 409:148–153. 2011. View Article : Google Scholar : PubMed/NCBI
36	Bustelo XR, Sauzeau V and Berenjeno IM: GTP-binding proteins of the Rho/Rac family: Regulation, effectors and functions in vivo. Bioessays. 29:356–370. 2007. View Article : Google Scholar : PubMed/NCBI
37	Liang L, Li Q, Huang LY, Li DW, Wang YW, Li XX and Cai SJ: Loss of ARHGDIA expression is associated with poor prognosis in HCC and promotes invasion and metastasis of HCC cells. Int J Oncol. 45:659–666. 2014. View Article : Google Scholar : PubMed/NCBI
38	Yamaguchi H and Condeelis J: Regulation of the actin cytoskeleton in cancer cell migration and invasion. Biochim Biophys Acta. 1773:642–652. 2007. View Article : Google Scholar : PubMed/NCBI
39	Maciver SK and Hussey PJ: The ADF/cofilin family: Actin-remodeling proteins. Genome Biol. 3:reviews30072002. View Article : Google Scholar : PubMed/NCBI
40	Aledo JC: Glutamine breakdown in rapidly dividing cells: Waste or investment? Bioessays. 26:778–785. 2004. View Article : Google Scholar : PubMed/NCBI
41	Le Bacquer O, Laboisse C and Darmaun D: Glutamine preserves protein synthesis and paracellular permeability in Caco-2 cells submitted to ‘luminal fasting’. Am J Physiol Gastrointest Liver Physiol. 285:G128–G136. 2003. View Article : Google Scholar : PubMed/NCBI
42	Spanaki C, Kotzamani D and Plaitakis A: Widening spectrum of cellular and subcellular expression of human Glud1 and Glud2 glutamate dehydrogenases suggests novel functions. Neurochem Res. 42:92–107. 2017. View Article : Google Scholar : PubMed/NCBI
43	Biankin AV, Waddell N, Kassahn KS, Gingras MC, Muthuswamy LB, Johns AL, Miller DK, Wilson PJ, Patch AM, Wu J, et al: Pancreatic cancer genomes reveal aberrations in axon guidance pathway genes. Nature. 491:399–405. 2012. View Article : Google Scholar : PubMed/NCBI
44	Gu C, Rodriguez ER, Reimert DV, Shu T, Fritzsch B, Richards LJ, Kolodkin AL and Ginty DD: Neuropilin-1 conveys semaphorin and VEGF signaling during neural and cardiovascular development. Dev Cell. 5:45–57. 2003. View Article : Google Scholar : PubMed/NCBI
45	Xu J and Xia J: NRP-1 silencing suppresses hepatocellular carcinoma cell growth in vitro and in vivo. Exp Ther Med. 5:150–154. 2013. View Article : Google Scholar : PubMed/NCBI
46	Pasterkamp RJ, Peschon JJ, Spriggs MK and Kolodkin AL: Semaphorin 7A promotes axon outgrowth through integrins and MAPKs. Nature. 424:398–405. 2003. View Article : Google Scholar : PubMed/NCBI
47	Monvoisin A, Neaud V, Lédinghen V, Dubuisson L, Balabaud C, Bioulac-Sage P, Desmoulière A and Rosenbaum J: Direct evidence that hepatocyte growth factor-induced invasion of hepatocellular carcinoma cells is mediated by urokinase. J Hepatol. 30:511–518. 1999. View Article : Google Scholar : PubMed/NCBI
48	Scagliotti GV, Novello S and von Pawel J: The emerging role of MET/HGF inhibitors in oncology. Cancer Treat Rev. 39:793–801. 2013. View Article : Google Scholar : PubMed/NCBI