MicroRNAs as possible biomarkers for diagnosis and prognosis of hepatitis B- and C-related-hepatocellular-carcinoma

Table 3 miRNAs patterns in studies enrolling hepatocellular carcinoma patients with hepatitis B virus and hepatitis C virus-related infection

Ref.	Characteristics of the study	miRNAs Up-regulated	miRNAs Down-regulated	Conclusions
Chung GE, Oncol Rep, 2010 Korea Period: 2001-2004	Tissue samples obtained from twenty-five pairs of primary HCC (18 HBV positive patients, 2 HCV positive subjects, 5 HBV/HCV negative) and adjacent non-tumor liver tissues were evaluated in this study	Up-regulation of miR-15b, miR-105 and miR-339, let-7d, miR-107, miR-103, miR-210, miR-25, let-7a, miR-93, miR-345, miR-30d, miR-423, miR-320	miR-422b, miR-22, miR-497, miR-195, miR-199a*, miR-199ª, miR-130a	miR-15b expression in HCC tissues may predict a low risk of HCC recurrence. In addition, the modulation of miR-15b expression may be useful as an apoptosis-sensitizing strategy for HCC treatment
Cong N, Tumor Biol, 2014 China Period: January 2007 - February 2012	Serum samples obtained from: -206 patients with HCC -217 controls	NR	NR	The miR-146a GG genotype and G allele carried an increased risk of HCC HBV-positive subjects carrying but not in HCV-infected patients
Coulouarn C, Oncogene, 2009 USA Period: NR	Specimens obtained from 64 HCC tissues (18 pts HBsAg +, 13 HCV +, 3 with HBV and HCV coinfection, 30 with different etiologies) and 28 matched non-tumor surrounding liver tissues from patients undergoing partial hepatectomy as treatment for HCC	NR	Down-regulation of miRNA-122 in HCC	miR-122 as a diagnostic and prognostic marker for HCC progression
Diaz G, Int J Cancer, 2013 Italy Period: NR	Tissue samples obtained from: -HCV-associated HCC (HCC), -HCC-associated non-tumours cirrhosis (HCC-CIR), -HCV- associated cirrhosis without HCC (CIR), -HBV-associated acute liver failure (ALF), -normal liver tissue surrounding angioma (NL), -normal liver from liver donors (LD)	Up-regulation of: miR 221, miR-224 and miR-224-3p, miR-452, miR-1269	Down-regulation of: miR-125a-5p , miR-130a, miR-139-5p, miR-139-3p, miR-195, miR-199a-5 and miR-199a-3p, miR-214, miR-424-3p, miR-497	18 miRNAs exclusively expressed in HCV-associated HCC and characterized by high specificity and selectivity vs all other liver diseases and healthy conditions were identified Among the 18 HCC-exclusive miRNAs identified in this research, miR-221 and miR-224, miR-199a-5p, miR-195, miR-214, miR-199a-3p, miR-125a-5p, miR-139-5p, miR-130a, miR-199b-3p, miR-139-3p, miR-224-3p and miR-452 were already reported in previous studies miR-497, miRNA-1269 miR-424-3p were never described in previous reports
Gramantieri L, Cancer Research, 2007 Italy Period: NR	Tissue obtained from: 60 patients (HBV: 5, HCV: 31, HBV+ HCV: 5, HCV+ pas tHBV: 5, past HBV: 1; HBV + ethanol: 1, HCV + ethanol: 2, ethanol: 3)	Up-regulation of: miR-221	Down-regulation of : let-7a-1, let-7a2, let-7a3, let-7b, let-7c, let-7d, let-7e, let-7f2, let-7g, miR-122a, miR-124a, miR-130a, miR-132, miR-136, miR-141, miR-142, miR-143, miR-145, miR-146, miR-150, miR-155, miR-181a-1, miR-181a-2, miR-181c, miR-195, miR-199a1-5p, miR-199a2-5p, miR-199b, miR-200b, miR-214, miR-223	The aberrant expression of a restricted panel of miRNAs could participate in the molecular events leading to HCC development
Hao YX, Asian Pac J Cancer Prev, 2013 China Period: January 2010 - April 2012	Serum samples from: -285 patients with HCC 133 HBsAg-positive 36 anti-HCV positive 8 with coinfection -Residents without HCC who entered the hospital for health check-ups were enrolled into control group Each control was pair-matched by sex and age (± 5 yr) to a patient with HCC	NR	NR	miR-196a2 CC genotype and C allele have an important role in HCC risk in Chinese patients, especially in HBV infection carriers No significant association observed between miR-146aG>C and miR-499A>G genetic polymorphisms and HCC risk
Köberle V, Eur J Cancer, 2013 Germany Period: February 2009 - July 2012	Serum samples obtained from: 195 patients with HCC ( 33 HBV+; 87 HCV +, 14 NASH, 65 Alcohol, 8 Haemochromatosis, 9 Cryptogenic) 54 patients with liver cirrhosis (2 HBV +; 41 HCV +, 0 NASH, 16 Alcohol, 0 Haemochromatosis, 0 Cryptogenic)	Longer OS in patients with higher miR-1 and miR-122 serum levels	Reduced OS in patients with lower miR-1 and miR-122 serum levels	At age-, sex-, tumor stage and treatment-adjusted multivariate Cox regression analysis miR-1 serum levels were independently associated with OS, whereas serum miR-122 was no. t miR-1 may improve the predictive value of classical HCC staging scores
Ladeiro Y, Hepatology, 2008 France Period: 1992-2004	109 liver samples, collected from 93 patients surgically treated. Analysed cases: HCC: 28, HC: 13, FNH: 5, non-tumor liver samples: 4 Two sets of samples were considered:(first set of samples (n = 50, 16 HBV positive, 9 HCV positive ) and validation set of samples (n = 59, 18 HBV positive, 17 HCV positive)	- miR-96 overexpressed in HBV tumors - miR-21, miR-222, miR-10b overexpression in HCC - miR-224 overexpression in HCC vs benign tumours	-miR-422b, miR-122a down-regulation both in benign and malignant tumors - miR-200c and miR-203 underexpression in benign tumours - low expression of miR-375 in both HCA and HCC mutated for β-catenin	Hepatocellular tumors may have a distinct miRNA expression fingerprint according to malignancy, risk factors, and oncogene/tumor suppressor gene alterations
Liu YX, BioMed Research International, 2014 China Period: January 2004 - December 2008	Tissue obtained from: -207 HCC liver tissue and patients and adjacent noncancerous tissue samples. HBV + : 174 patients; HCV+ : 3 patients	miR-24 up-regulation in HCC tumor tissues relative to adjacent noncancerous tissue samples	NR	High expression of miR-24 could promote AFB1-DNA formation and increase adducts mount. High expression of miR-24 was significantly correlated with larger tumor size, higher microvessel density, and tumor dedifferentiation
Liu WH, Gastroenterology, 2009 Taiwan Period: 2002-2006	Tissue obtained from: -80 HCC patients (40 HBV+ and 40 HCV +), -16 focal nodular hyperplasia cases -7 adenoma cases	Specifically increased miR-18a miRNA in samples from female HCC patients. miR-18a expression in tumor tissues not different from the non-tumoral tissues in either male or female patient FNHs and adenomas	NR	miR-18a prevents translation of ER, potentially blocking the protective effects of oestrogen and promoting the development of HCC in women
Lu CY, Genes Chromosomes and Cancer, 2013 Taiwan Period: NR	Tissue and sera obtained from: -41 patients with HCC (19 HBV positive, 11 HCV positive, 2 HBV/HCV positive, 8 HBV/HCV negative and 1 not determined) -8 patients with cirrhosis (6 HBV positive, 2 HCV positive, 1 HBV/HCV positive), -10 Healthy subjects	In 39/41 HCC, the methylation levels of miR-129-2 were significantly increased in tumor tissues compared with adjacent normal tissues miR-129-2 methylation was detectable in plasma samples from HCC patients, but not in plasma samples from healthy individuals or patients with liver cirrhosis	NR	miR-129-2 methylation is highly accurate in distinguishing HCC patients from cirrhosis patients and healthy individuals, implying its potential utility as an early diagnostic marker for HCC
Murakami Y, Oncogene, 2006 Japan Period: NR	miR expression profiles in 25 pairs of hepatocellular carcinoma (HCC) and adjacent nontumorous tissue (NT) and nine additional CHB specimens was performed, using a human miRNA microarray HBV +: 6; HCV +: 26	Major expression of miR-18, precursor miR-18, and miR-224 in HCC vs non-cancerous tissue	Minor expression of miR-199a*, miR-195, miR-199a, miR-200a, and miR-125ª in HCC vs non-cancerous tissue	Higher expression of three miRNAs in the HCC samples vs NT samples, demonstrated lower expression of five miRNAs in the HCC samples vs NT samples
Qu KZ, J Clin Gastroenterol, 2011 USA Period: NR	283 subjects studied: -105 patients with HCC (20 HBV +, 66 HCV +, 1 with coinfection); -107 individuals with CLDs; -71 healthy controls	NR	Significantly lower serum levels of miR-16 and miR-199a in HCC than in CLD patients or control subjects	Measurement of serum levels of miR-16 improves differentiation of HCC from non-HCC CLD. The combination of miR-16, AFP, AFP-L3, and DCP yielded greater sensitivity and specificity for HCC detection than any other single marker or marker combination examined
Salvi A, Intern J Oncol, 2012 Italy Period: NR	Tissue specimens obtained from: human HCC samples, corresponding peritumoral and non-tumor samples (resected 1-2 cm from the malignant tumor) 15 HCV + patients, 10 HBV+ patients 4 HBV+/HCV + subjects 7 HBV -/ HCV - patients 5 patients with no available informations (25 cirrhosis, 15 hepatitis, 1 steatosis)	Up-regulation of: miR-21 in HCC tissues vs the corresponding peritumoral tissues, particularly in non-cirrhotic HCC	Down-regulation of: miR-24 and miR-27a in HCCs from cirrhotic liver tissues in comparison to those from non-cirrhotic liver tissues. The downregulation of miR-24 was correlated with poorer prognosis in patients with HBV and HCV virus infections	Differential expression of miRNAs in cirrhotic and non-cirrhotic HCCs, thereby contributing to advances in the discovery and validation of novel molecular biomarkers of HCC progression
Sato F, PLoS One, 2011 Japan Period: January 1997 - March 2007	73/639 patients with HCC and satisfying enrollment criteria, underwent hepatic resection Patients HBsAg +: 12; Patients HCV +: 51	Recurrence-related miR in tumor tissues: miR22, miR99a, miR99b, miR100, miR 125a-5p, miR125b, miR129-5p, miR 140-3p, miR145, miR195, miR221, miR378, miR497	Recurrence-related miRNA in non-tumor tissues: miR18a, miR18b, miR21, miR23a, miR24, miR27a, miR96, miR103, miR 107, miR126, miR142- 3p, miR 148a, miR 191, miR 222, miR362-3p, miR 425, miR378, miR1202, let 7e, let-7f	miRNA profiling can predict HCC recurrence in Milan criteria cases miR-96 in non-tumor tissues is the most strongly associated with HCC recurrence
Shigoka M, Pathology International, 2010 Japan Period: NR	Serum and tissue samples obtained from: -22 HCC cases (6 HBV +, 10 HCV +, 6 non-HBV/HCV) -5 pairs of fresh HCC and non-tumorous LCD samples surgically resected from HCC patients -10 healthy subjects	Higher levels of miR-92a expression in HCC sections vs adjacent LC sections	Decreased ratio of miR- 92a to miR-638 in the plasma samples from the HCC patients compared with that from the normal donors	Deregulation of miR-92 expression in cells and plasma could be implicated in the development of HCC
Spaniel C, PLoS One, 2013 Japan/USA Period: NR	Tissue samples obtained from: a) Paired tumor and nontumor tissues collected from 26 patients undergoing surgical resection of HCC: -16 with concomitant chronic HCV infection, -10 infected with HBV b) 9 with non-infected ‘normal’ liver tissue collected from patients undergoing resection of metastases of non-hepatic primary cancers	Possible miR-191 increased expression in HBV-associated HCC	Significant reduction of miR-122 abundance in HBV associated HCC in comparison with “normal” liver tissue, but not in liver cancer associated with HCV. Significant differences in miR-122 expression exist in non-tumor tissue, with miR-122 abundance reduced from “normal” in HCV- but not HBV-infected liver	miR-122 abundance varies between HBV- and HCV-related liver HCC as well as in non-tumor tissue
Toffanin S, Hoshida Cabellos Gastroenterology, 2011 Italy Period: NR	Tissue samples obtained from: - 89 fresh-frozen HCC samples (surgical resection or LT); - Formalin-fixed paraffin-embedded tissues of 165 HCCs (validation set) caused by HCV, HBV, alcohol, and others Subjects subdivided into: Training set: 79 patients Validation set: 161 patients Training set:, HCV +: 79/79 patients Validation set: HCV +: 74/161; HBV +: 43/161; Alcohol: 12/161; Other: 25/161	Up-regulation of 23 miRNAs in cluster C2 (miR-517a, miR-517b, miR-517c, miR-520g, miR-520h, miR-519b, miR-519d, miR-516-5p, miR-519a,miR-520c, miR-520b, miR-520f, miR-526b, miR-524, miR-516-1, miR-526b, miR-519e, miR-512-3p, miR-522, miR-526a, miR-518f, miR-518b, and miR-525 Up-regulation of 16 miRNAsin cluster C3 (miR-376a, miR-494, miR-409-3p, miR-376b, miR-377, miR-368, miR-382, miR-369-3p, miR-410, miR-432, miR-154, miR-379, miR-299-5p, miR-431, miR-381, miR-495)	Down-regulation of miR-26a and miR-26b in cluster C2 and C3	miRNA-based classification of 3 subclasses of HCC is proposed. Among the proliferation class, miR-517a is an oncogenic miRNA, promoting tumor progression A rationale for developing therapies that miRNA 517 for patients with HCC is proposed A hierarchical clustering of miRNA data identified 3 main clusters of HCC: clusters A (32/89), B (29/ 89) and C (28/89). The C cluster divided into 3 sub-clusters with distinct miRNA expression patterns: C1 (15/89), C2 (8/89) and C3 (5/89)
Tomimaru Y, J Hepatol, 2011 Japan Period: January2010 - February 2010	Serum samples obtained from: -10 patients before and after curative resection of HCC (HBV/HCV-: 1, HBV: /3/ HCV+: 6/); -126 patients with HCC, -30 patients with CLD, -50 healthy volunteers	Significantly higher plasma miR-21 level in the HCC patients in comparison with CLD patients and healthy volunteers	Significantly diminished plasma miRNA-21 levels after surgery compared with the pre-operative values	Plasma miRNA-21 level is a promising biochemical marker for HCC
Ura S Hepatology, 2009 Japan Period: 1999-2004	12 patients with HBV-related HCC 14 patients with HCV-related HCC	NR	Commonly repressed miR in CH-B, CH-C, HCC-B, and HCC-C compared with normal liver: miR-219, miR-320, miR-154, miR-29c; miR-338; miR-26a; miR-126; miR-325	miRNAs as important mediators of HBV and HCV infection as well as liver disease progression, they could be potential therapeutic target molecules Major miRNAs expression in HCC vs CLD: miR21, miR-98, miR183, miR221, miR222, miR301. Minor miRNAs expression in HCC vs CLD: miR17-3p, miR30a-3p, miR30e, miR92, miR 99a, miR122, miR125b, miR130a, miR139, miR187, miR199a, miR200a, miR200b, miR223, miR326
Zhou B Tumor Biol, 2014 China Period: January 2010 - February 2012	Serum samples obtained from: -266 patients with HCC -281 Healthy controls	NR	NR	Subjects with miR-146a GG and G allele had an enhanced risk of HCC in comparison with homozygote CC genotype. Individuals with miR-196a2, TT and T allele significantly decreased the risk of HCC in comparison with CC genotype. miR-196a2C>T polymorphisms associated with a decreased risk of HBV-related HCC, but not in HCV-related HCC cases

ABH: Acute B hepatitis; ASCs: Asymptomatic HBsAg carriers; BCLC: Barcelona Clinic Liver Cancer staging system; CHB: Chronic hepatitis B; CLD: Chronic liver disease; ER-α: Estrogen receptor-α; FNH: Focal nodular hyperplasia; HCC: Hepatocellular carcinoma; HC: Healthy controls; HCA: Hepatocellular Adenoma; LC: Liver cirrhosis; LT: Liver transplantation; NR: Not reported; OS: Overall survival; RFS: Recurrence-free survival; SR: Spontaneously recovered.