MicroRNA profile in HBV-induced infection and hepatocellular carcinoma - PubMed (original) (raw)

MicroRNA profile in HBV-induced infection and hepatocellular carcinoma

Guanyu Wang et al. BMC Cancer. 2017.

Abstract

Background: MicroRNAs (miRNAs) exhibit essential regulatory functions related to cell growth, apoptosis, development and differentiation. Dysregulated expression of miRNAs is associated with a wide variety of human diseases. As such miRNA signatures are valuable as biomarkers for disease and for making treatment decisions. Hepatitis B virus (HBV) is a major risk factor for hepatocellular carcinoma (HCC). Here we screened for miRNAs in chronic HBV associated HCC.

Methods: To determine the miRNAs in HCC occurrence associated with HBV infection, we analyzed global miRNA expression profiles in 12 pairs of HCC and adjacent matched non-HCC tissues from HBV-positive and HBV-negative patients using microarray analyses. The microarray result was validated by real-time PCR in 32 HBV-positive and 24 HBV-negative patient HCC samples. The potential candidate target genes of the miRNAs were predicted by miRWalk software. Genes simultaneously predicted as targets by two or more miRNAs were subjected to GO and KEGG pathway analysis. The miRNA regulatory network analysis was performed using the Ingenuity Pathway Analysis (IPA) software.

Results: Eight miRNAs (miR-223, miR-98, miR-15b, miR-199a-5p, miR-19b, miR-22, miR-451, and miR-101) were involved in HBV-unrelated HCC, 5 miRNAs (miR-98, miR-375, miR-335, miR-199a-5p, and miR-22) were involved in HBV infection, and 7 miRNAs (miR-150, miR-342-3p, miR-663, miR-20b, miR-92a-3p, miR-376c-3p and miR-92b) were specifically altered in HBV-related HCC. Gene Ontology and KEGG analyses predict that these HBV-related HCC miRNAs are involved in the regulation of: transcription, RNA polymerase II promoter, phosphorylation of proteins through MAPK signaling pathway, focal adhesion, and actin cytoskeleton. IPA analysis also suggest that these miRNAs act on AGO2, TP53, CCND1, and 11 other genes that significantly influence HCC occurrence and HBV infection.

Conclusion: Our data indicates that the unique 7 miRNAs expression signature could be involved in the development HBV- related HCC.

Keywords: Hepatitis B virus; Hepatocellular carcinoma; Regulatory network; microRNA.

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Conflict of interest statement

This study was performed in strict accordance with the recommendations from the Guide for Clinical Research provided by Sir Run Run Shaw Hospital, Zhejiang University. The protocol was approved and monitored by the Ethics Committee of Sir Run Run Shaw Hospital, Zhejiang University. Signed informed consent was obtained from each patient.

Not applicable.

Competing interests

The authors declare that they have no competing interests.

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Figures

Fig. 1

Fig. 1

qPCR results of candidate miRNA confirm microarray data. a 5 miRNAs were selected for confirmation by qPCR. in HCC and ajdjacent non tumor tissues. b The expression of miR-22. c The expression of miR-98. d The expression of miR-155. e The expression of miR-361. f The expression of miR-223. PP = adjacent non tumor tissues of HBV-positive patients, TP = tumors of HBV-positive patients, PN = adjacent non-tumor tissues of HBV-negative patient, TN = tumors of HBV-negative patients

Fig. 2

Fig. 2

Venn diagram analysis of the relationships among target genes predicted by differentially expressed miRNAs in HBV infection. a Relationships among the target genes predicted by miR-98, miR-375, and miR-335. b Relationships among target genes predicted by miR-199a-5p and miR-22

Fig. 3

Fig. 3

GO and pathway analysis results of the target genes predicted by differentially expressed miRNAs in HBV infection. -log10(P) indicates the GO score related to genes with the biological process by _P_-value. a Main biological processes influenced by genes targeted by two or more miRNAs (miR-98, mir-375, and miR-335). b Main biological processes influenced by genes targeted by miR-199a-5p, and miR-22. c Main pathway influenced by genes targeted by two or more miRNAs from miR-98, mir-375, and miR-335. d Main pathway influenced by genes targeted by miR-199a-5p, and miR-22

Fig. 4

Fig. 4

Venn diagram analysis of the relationship among target genes predicted by differentially expressed miRNAs which are involved in HBV-induced HCC. a Relationship among target genes predicted by miR-20b, miR-92a-3p, miR-92b, and miR-376c-3p. b Relationship among target genes predicted by miR-150, miR-342-3p, and miR-663

Fig. 5

Fig. 5

GO and pathway analysis result of the target genes predicted by differentially expressed miRNAs which are involved in HBV-induced HCC. -log10(P) indicates the GO score related to genes in the biological process by _P_-value. a Main biological processes influenced by genes targeted by two or more miRNAs from miR-20b, miR-92a-3p, miR-92b, and miR-376c-3p. b Main biological processes influenced by genes targeted by two or more miRNAs from miR-150, miR-342-3p, and miR-663. c Main pathways influenced by genes targeted by two or more miRNAs from miR-20b, miR-92a-3p, miR-92b, and miR-376c-3p. d Main pathways influenced by genes targeted by two or more miRNAs from miR-19b, miR-101, and miR-199a-5p

Fig. 6

Fig. 6

Results of IPA analysis of the target genes predicted by differentially expressed miRNAs involved in HBV-indcued HCC. IPA-obtained network showing the relationships among 6 co-regulated miRNAs (miR-150, miR-342-3p, miR-92a-3p, miR-92b, and miR-376c-3p)

References

    1. Kerr TA, Korenblat KM, Davidson NO. MicroRNAs and liver disease. Transl Res. 2011;157(4):241–252. doi: 10.1016/j.trsl.2011.01.008. - DOI - PMC - PubMed
    1. Motavaf M, Safari S, Saffari Jourshari M, Alavian SM. Hepatitis B virus-induced hepatocellular carcinoma: the role of the virus x protein. Acta Virol. 2013;57(4):389–396. doi: 10.4149/av_2013_04_389. - DOI - PubMed
    1. Su IJ, Wang HC, Wu HC, Huang WY. Ground glass hepatocytes contain pre-S mutants and represent preneoplastic lesions in chronic hepatitis B virus infection. J Gastroenterol Hepatol. 2008;23(8 Pt 1):1169–1174. doi: 10.1111/j.1440-1746.2008.05348.x. - DOI - PubMed
    1. Soussan P, Garreau F, Zylberberg H, Ferray C, Brechot C, Kremsdorf D. In vivo expression of a new hepatitis B virus protein encoded by a spliced RNA. J Clin Invest. 2000;105(1):55–60. doi: 10.1172/JCI8098. - DOI - PMC - PubMed
    1. El-Serag HB, Mason AC, Key C. Trends in survival of patients with hepatocellular carcinoma between 1977 and 1996 in the United States. Hepatology. 2001;33(1):62–65. doi: 10.1053/jhep.2001.21041. - DOI - PubMed

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