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microRNAs (miRs) are 18-25 nucleotide non-coding RNAs that regulate geneexpression in several physiological and pathological conditions. To gather moreknowledge on microRNAs in human hepatocellular carcinoma (HCC) we generated asmall RNA library in the human HCC cell line HA22T/VGH by cloning and sequencingthe cDNA obtained following the size selection of 18-24 nucleotide RNAs. We determinedthe expression levels of the most frequently cloned microRNAs by qPCR in HCC tissuesand in their peritumoral counterparts from biopsy specimens of 41 HCC patients.The most frequently cloned miRs were miR-24, miR-27a and miR-21, and their expressionlevels in human HCC tissues indicate that these miRs were dysregulated in HCC.We showed that miR-24 and miR-27a were significantly downregulated in HCCs fromcirrhotic liver tissues in comparison to those from non-cirrhotic liver tissues.In cirrhotic HCCs the downregulation of miR-24 was correlated with poorer prognosisin patients with HBV and HCV virus infections. miR-21 was generally upregulatedin HCC tissues versus the corresponding peritumoral tissues, particularly in non-cirrhoticHCC. Furthermore, by sequence alignment we identified the human miR orthologueof Mus musculus miR-1199 not yet annotated. Our results outline the differentialexpression of miRs in cirrhotic and non-cirrhotic HCCs, thereby contributing toadvances in the discovery and validation of novel molecular biomarkers of HCCprogression.

Human hepatocellular carcinoma cell-specific miRNAs reveal the differential expression of miR-24 and miR-27a in cirrhotic/non-cirrhotic HCC