Genomic instability in chronic viral hepatitis and hepatocellular carcinoma

doi:10.1053/hupa.2001.25593

Human Pathology

Volume 32, Issue 7, July 2001, Pages 698-703

https://doi.org/10.1053/hupa.2001.25593 Get rights and content

Abstract

Chronic hepatitis may progress to cirrhosis and hepatocellular carcinoma (HCC). Progressive accumulation of mutations and genomic instability in chronic viral hepatitis might flag an increased risk of HCC development. Genomic instability at dinucleotide microsatellite loci in chromosomes 2, 13, and 17 and at 2 mononucleotide repeat loci was examined in liver tissues from 41 patients, including 30 without HCC (18 patients with chronic hepatitis and 12 with cirrhosis) and 11 with HCC. Genomic instability was detected in 51% of the 41 cases. Allelic imbalance at informative dinucleotide loci occurred in 37% of the cases. In 14 cases (34%), allelic imbalance was detected in chronic hepatitis or cirrhosis without HCC. Allelic imbalance at the chromosome 13 locus was detected in 50% of the cases of chronic hepatitis C. Allelic imbalance at the TP53 chromosome locus and/or at the chromosome 13 locus was significantly more frequent than alterations at the chromosome 2 locus (P =.026). Low-level microsatellite instability was found in 20% of all cases examined and high-level microsatellite instability in 3 patients (7.5%), including 2 cases of chronic hepatitis and 1 case of cirrhosis. Our results show that allelic imbalance occurs frequently in hepatitis-related HCC as well as in chronic hepatitis in patients without HCC. Allelic imbalance at the D13S170 chromosome 13 locus (13q31.2) occurs frequently in chronic hepatitis, suggesting that genomic alterations affecting the long arm of chromosome 13 might be used to monitor the natural progression of chronic hepatitis-associated liver carcinogenesis. HUM PATHOL 32:698-703. Copyright © 2001 by W.B. Saunders Company

Section snippets

Tissue sample collection

Liver samples obtained from patients with HCV-related chronic liver disease and HCC were used. Liver tissue was obtained by percutaneous or wedge liver biopsy or by surgical resection at the Institutes of Clinica Medica and Clinica Chirurgica, University of Medicine, Sassari, Italy. A total of 41 samples were analyzed, including 11 surgical resection specimens and 30 needle biopsies. Eighteen patients had a histologic diagnosis of chronic hepatitis without cirrhosis, 12 had a diagnosis of

Results

Forty-one patients were studied (mean + SD age, 54.9 ± 15 years; range, 19 to 78 years). There were 17 women and 24 men (Table 1). All patients were white Italians from the same geographical region. Sera of all patients were anti-HCV and HCV RNA positive. Sixteen patients were also positive for anti-HBc immunoglobulin (Ig)G but negative for antibody to hepatitis B surface antigen (anti-HBsAg) IgG and for HBV DNA in the serum.

Chronic hepatitis grading and staging was performed according to

Discussion

In this study, we found that AI affecting the hepatic parenchyma was a frequent event in chronic viral hepatitis, providing a possible molecular mechanism for the progression of hepatic carcinogenesis associated with chronic hepatitis. We identified patients with hepatitis or cirrhosis who did not have evidence of carcinoma but nonetheless displayed obvious genetic anomalies that might predispose to cancer development.

MSI was infrequent, and high-level MSI was detected in only 2 cases of

References (37)

RD Kolodner et al.
Eukaryotic DNA mismatch repair
Curr Opin Genet Dev
(1999)
WK Leung et al.
Microsatellite instability in gastric intestinal metaplasia in patients with and without gastric cancer
Am J Pathol
(2000)
Y Kondo et al.
Microsatellite instability associated with hepatocarcinogenesis
J Hepatol
(1999)
M Roncalli et al.
Comprehensive allelotype study of hepatocellular carcinoma
Hepatology
(2000)
S Tamura et al.
Association of cumulative allelic losses with tumor aggressiveness in hepatocellular carcinoma
J Hepatol
(1997)
YW Lin et al.
Loss of heterozygosity at chromosome 13q in hepatocellular carcinoma: identification of three independent regions
Eur J Cancer
(1999)
M Colombo
Hepatitis C virus and hepatocellular carcinoma
Semin Liver Dis
(1999)
RP Beasley
Hepatitis B virus. The major etiology of hepatocellular carcinoma
Cancer
(1988)
I Saito et al.
Hepatitis C virus infection is associated with the development of hepatocellular carcinoma
Proc Natl Acad Sci U S A
(1990)
JD Groopman et al.
Molecular biomarkers for aflatoxins: From adducts to gene mutations to human liver cancer
Can J Physiol Pharmacol
(1996)

PC Nowell

Mechanisms of tumor progression

Cancer Res

(1986)

D Lasko et al.

Loss of constitutional heterozygosity in human cancer

Annu Rev Genet

(1991)

E Solomon et al.

Chromosome aberrations and cancer

Science

(1991)

GA MacDonald et al.

Microsatellite instability and loss of heterozygosity at DNA mismatch repair gene loci occurs during hepatic carcinogenesis

Hepatology

(1998)

LA Aaltonen et al.

Clues to the pathogenesis of familial colorectal cancer

Science

(1993)

A Sepulveda et al.

Marked differences in the frequency of microsatellite instability in gastric cancer from different countries

Am J Gastroenterol

(1998)

G Sozzi et al.

Detection of microsatellite alterations in plasma DNA of non-small cell lung cancer patients: A prospect for early diagnosis

Clin Cancer Res

(1999)

JT Johannsdottir et al.

The effect of mismatch repair deficiency on tumourigenesis: Microsatellite instability affecting genes containing short repeated sequences

Int J Oncol

(2000)

Cited by (31)

Polymorphisms in hepatocellular carcinoma
2022, Theranostics and Precision Medicine for the Management of Hepatocellular Carcinoma, Volume 1: Biology and Pathophysiology
Liver is the major glandular organ in the human body involved in homeostasis, detoxification, and metabolism. Different cells in the liver presented with distinct function sometimes lead to the formation of either benign or malignant liver tumors. Precisely 5%–10% of all cancers are due to genetic alterations. Hepatocellular carcinoma cancer is not exception in this regard. Liver cancer (LC) is one of the other cancers causing major burden in terms of prevalence and death in the world. Often, the death rate in patients with LC is elevated due to its late occurrence in advanced stages. The risk factors include age, gender, food habits, lifestyle, viral infections, and genetic aberrations. The risk of LC doubled with the family history of cancer. Various genetic alterations include chromosomal instability, microsatellite instability, and single-nucleotide polymorphisms (SNP) that are the main factors in the development of LC. Reports suggest that there are specific SNP, like rs17401966, which are highly associated with LC. Various mutations identified in the genes TP53, catenin, beta 1, Axin1, p16INK4, insulin-like growth factor 2 receptor, RB transcriptional corepressor 1, and cyclin D1 involved in multiple pathways are important for cell signaling and apoptosis. In addition to mutations, the loss of heterozygosity was identified at various loci in chromosomes 1, 4, 9, 13, and 17. As the death rate is more in LC, it is highly demanding to put our research into the study of genes involved in liver carcinogenesis.
Identification of NUCKS1 as a putative oncogene and immunodiagnostic marker of hepatocellular carcinoma
2016, Gene
Citation Excerpt :
Amplification of chromosome 1q is one of the most frequent genetic alterations in primary HCC, and can be detected in 58–78% of HCC patients (Marchio et al., 1997; Kusano et al., 1999; Wong et al., 1999; Guan et al., 2000). Another previous report showed that allelic imbalance occurs frequently in chronic hepatitis patients without HCC as well as in chronic hepatitis-associated HCC (Dore et al., 2001). This same study showed that copy number alterations affecting the chromosome 13q might be used to monitor the progression of chronic hepatitis-associated hepatocarcinogenesis.
Although the molecular mechanisms underpinning hepatocellular carcinoma (HCC) are unknown, gene copy number and associated mRNA expression changes are frequently reported. Comparative genomic hybridization arrays spotted with 4041 bacterial artificial chromosome clones were used to assess copy number changes in 45 HCC tissues. Seventy more HCC tissues were used to validate candidate genes by using western blots and immunohistochemistry. A total of 259 clones were associated with copy number changes that significantly differed between normal liver and HCC samples. The chromosomal region 1q32.1 containing the nuclear casein kinase and cyclin-dependent kinase substrate 1 (NUCKS1) gene was associated with tumor vascular invasion. Western blot analysis demonstrated that NUCKS1 was up-regulated in 37 of 70 (52.8%) HCC tissues compared with adjacent non-tumor tissues, and over-expressed in a vast majority of HCCs (44/52, 84.6%) as determined by immunohistochemical staining. Furthermore, immunostaining of both NUCKS1 and glypican-3 improved the diagnostic prediction of HCC. Knock-down of NUCKS1 by siRNA implied the decrease in cell viability of the Hep3B cell line and reduced tumor formation in a xenograft mouse model. NUCKS1 was identified as a potential oncogene at chromosomal 1q32.1 in patients with HCC, and it might be a valuable immunodiagnostic marker for HCC.
Pathological and molecular analysis of sporadic hepatic angiomyolipoma
2006, Human Pathology
Hepatic angiomyolipoma (HAML) is an unusual mesenchymal lesion that can be misdiagnosed as hepatocellular carcinoma or sarcoma. We sought to better define the morphological variations and immunohistochemical and molecular features of this unusual tumor. Forty-nine sporadic HAMLs were investigated for immunopathologic characteristics with EnVision Plus. The clonal analysis was based on the methylation pattern of the polymorphic X chromosome–linked human androgen receptor gene, loss of heterozygosity (LOH), and microsatellite instability (MSI) detected with polymerase chain reaction using the MegaBACE 500 automatic system on microdissected tissues. Histologically, HAML is composed of a heterogeneous mixture of blood vessels, smooth muscle, and adipose cells. The myomatous or myoid cells were the most variable. Most of the tumor cells were positive for HMB-45 (100%) and SMA (100%). There was a typical monoclonal pattern in 35 of the 40 tumors. No LOH or MSI was found. Hepatic AML is a benign neoplasm with varied morphology and monoclonal growth. HMB-45 is the best marker available for diagnosis. Neither LOH nor MSI appears to play an important role in the pathogenesis of this tumor.
Hepatitis C virus NS5B delays cell cycle progression by inducing interferon-β via Toll-like receptor 3 signaling pathway without replicating viral genomes
2006, Virology
Citation Excerpt :
Furthermore, we found that the core protein promoted microsatellite instability in PH5CH8 cells (Naganuma et al., 2004). In fact, microsatellite instability was detected in approximately 20% of the tumor tissues from HCC patients examined, whereas no microsatellite instability was detected in normal liver tissues from the same patients (Dore et al., 2001; Kondo et al., 2000). In order to clarify the effect of HCV proteins on cell cycle progression in PH5CH8 cells, we examined cell cycle progression after the cells were released from the G1/S boundary in PH5CH8 cells expressing HCV proteins.
To clarify the pathogenesis of hepatitis C virus (HCV), we have studied the effects of HCV proteins using human hepatocytes. Here, we found that HCV NS5B, an RNA-dependent RNA polymerase, delayed cell cycle progression through the S phase in PH5CH8 immortalized human hepatocyte cells. Since treatment with anti-interferon (IFN)-β neutralizing antibody restored the cell cycle delay, IFN-β was deemed responsible for the cell cycle delay in NS5B-expressing PH5CH8 cells. The induction of IFN-β and the cell cycle delay were overridden by the down-regulation of Toll-like receptor 3 (TLR3) through RNA interference in NS5B-expressing PH5CH8 cells. Moreover, the NS5B full form was required for the cell cycle delay, the induction of IFN-β, and the activation of the IFN-β signaling pathway. Our findings revealed that NS5B induced IFN-β through the TLR3 signaling pathway in immortalized human hepatocytes even without replicating viral genomes.
Allelic alterations in nontumorous liver tissues and corresponding hepatocellular carcinomas from chinese patients
2003, Human Pathology
Allelic imbalance may play an important in tumor progression in hepatocarcinogenesis, but the genetic background of the corresponding nontumorous liver in hepatocellular carcinoma (HCC) is not well defined. We studied the incidence of loss of heterozygosity (LOH) and microsatellite instability (MSI) by microsatellite analysis in both nontumorous livers and the corresponding tumors, by comparing them with the normal DNA from Chinese patients with resected primary HCCs. We also evaluated the pathologic significance of the alterations. We used 18 highly polymorphic microsatellite markers on chromosomes 1, 3, 4, 7, 8, 9, 13, 16, 17, and 18. Our results showed that 70.6% (24 of 34) of the HCCs exhibited LOH at 1 or more loci, and that the overall fractional allelic loss (FAL) was 0.169. MSI was observed in only 1 tumor. In contrast, the nontumorous livers of the HCCs showed a very low incidence of LOH, with only a single LOH detected in 1 of 34 (2.9%) of the nontumorous livers, with an overall FAL index of 0.005. Tumors with LOH at 1 or more loci had significantly more frequent venous invasion (P = 0.019). Allelic loss at locus D9S199 (9p23) was seen more frequently in larger tumors (P = 0.031), and, less significantly, allelic loss at locus D16S516 (16q24.1) was seen more frequently in larger tumors (P = 0.059). LOH was common in predominantly hepatitis B virus-associated HCCs from Chinese patients. However, LOH or MSI in the corresponding cirrhotic or noncirrhotic livers was uncommon.
Development and Validation of Genome Instability-Associated lncRNAs to Predict Prognosis and Immunotherapy of Patients With Hepatocellular Carcinoma
2022, Frontiers in Genetics

View all citing articles on Scopus

^☆: Supported by the Department of Veterans Affairs; by the generous support of Hilda Schwartz, Houston, TX; and by a grant from the Ministero dell'Università e della Ricerca Scientifica e Tecnologica (MURST), Rome, Italy.

^☆☆: Address correspondence and reprint requests to Antonia R. Sepulveda, MD, PhD, University of Pittsburgh Medical Center, Department of Pathology PUH-A610, 200 Lothrop St, Pittsburgh, PA 15213-2582.

View full text

Original ContributionsGenomic instability in chronic viral hepatitis and hepatocellular carcinoma☆,☆☆

Abstract

Section snippets

Tissue sample collection

Results

Discussion

Curr Opin Genet Dev

Am J Pathol

J Hepatol

Hepatology

J Hepatol

Eur J Cancer

Hepatitis C virus and hepatocellular carcinoma

Semin Liver Dis

Hepatitis B virus. The major etiology of hepatocellular carcinoma

Cancer

Hepatitis C virus infection is associated with the development of hepatocellular carcinoma

Proc Natl Acad Sci U S A

Molecular biomarkers for aflatoxins: From adducts to gene mutations to human liver cancer

Can J Physiol Pharmacol

Mechanisms of tumor progression

Cancer Res

Loss of constitutional heterozygosity in human cancer

Annu Rev Genet

Chromosome aberrations and cancer

Science

Microsatellite instability and loss of heterozygosity at DNA mismatch repair gene loci occurs during hepatic carcinogenesis

Hepatology

Clues to the pathogenesis of familial colorectal cancer

Science

Marked differences in the frequency of microsatellite instability in gastric cancer from different countries

Am J Gastroenterol

Detection of microsatellite alterations in plasma DNA of non-small cell lung cancer patients: A prospect for early diagnosis

Clin Cancer Res

The effect of mismatch repair deficiency on tumourigenesis: Microsatellite instability affecting genes containing short repeated sequences

Int J Oncol

Original Contributions
Genomic instability in chronic viral hepatitis and hepatocellular carcinoma☆,☆☆