Detection of hypermethylation of the p16INK4A gene promoter in chronic hepatitis and cirrhosis associated with hepatitis B or C virus

Abstract

Background/aim-Inactivation of the p16 INK4A (p16) tumour suppressor gene by promoter region hypermethylation has been demonstrated not only in many types of tumours, including hepatocellular carcinoma (HCC), but also in early preneoplastic lesions in the lung, colon, oesophagus, and pancreas. The aim of this study was to examine the methylation status of the p16 promoter in pre-and/or non-neoplastic liver diseases. Patients/subjects/methods-The methylation status of p16 was evaluated in 22 HCC, 17 cirrhosis, 17 chronic hepatitis, nine primary biliary cirrhosis (PBC), eight autoimmune hepatitis, seven drug induced liver disease, six fatty liver, and three normal liver tissues using methylation specific polymerase chain reaction (MSP). p16 protein expression was also examined by immunohistochemical staining. Results-Methylation

Methylation of multiple genes as molecular markers for diagnosis of a small, well‐differentiated hepatocellular carcinoma

“…[5][6][7] Clinical studies investigating associations between carcinogenesis and abnormal methylation on CpG islands in tumor suppressor genes or genes involved in cell proliferation and death have been conducted in various cancers. [8][9][10][11][12] For HCC, it has been reported that some genes undergo hypermethylation in liver tissues, [13][14][15][16][17][18][19][20][21] supporting the hypothesis that determination of methylation in specific genes may be useful for HCC diagnosis. However, to the best of our knowledge, the diagnosis of HCC, in particular early HCC, based on the methylation levels of a single gene has not been clinically investigated to date.…”

“…APC, CASP8, CCND2, CFTR, CDKN2A, GSTP1, HIC1, POU3F1, RASSF1A, RUNX3, SPINT2 and TP73 were the genes filtered and were selected from previously reported data. [13][14][15][16][17][18][19][20][21] With the exception of CASP8, all genes were found to contain CpG islands located in their promoter regions by CpG mapping analysis, and the regions including their short CpG-rich stretches were successfully amplified for methylation analysis by sequencing. For CASP8, the region located in intron 3 that contained several CpG dinucleotide sequences was amplified (Table II).…”

“…However, to the best of our knowledge, the diagnosis of HCC, in particular early HCC, based on the methylation levels of a single gene has not been clinically investigated to date. [13][14][15][16][17][18][19][20][21] The aim of the current study was to identify a combination of methylated marker genes suitable for use in the diagnosis of a small HCCs less than 3 cm HCC or a well-differentiated HCC as an early HCC, and to establish a user-friendly methylation-specific PCR (MSP) system to measure methylation status of these HCC gene markers. To achieve the aim, we selected 12 genes (APC, CASP8, CCND2, CFTR, CDKN2A, GSTP1, HIC1, POU3F1, RASSF1A, RUNX3, SPINT2 and TP73) with a clear methylation in >50% (actually 68-100%) of HCC cases examined in studies reported previously as genetic marker candidates for diagnosis of early HCC.…”

“…To achieve the aim, we selected 12 genes (APC, CASP8, CCND2, CFTR, CDKN2A, GSTP1, HIC1, POU3F1, RASSF1A, RUNX3, SPINT2 and TP73) with a clear methylation in >50% (actually 68-100%) of HCC cases examined in studies reported previously as genetic marker candidates for diagnosis of early HCC. [13][14][15][16][17][18][19][20][21] We then analyzed the methylation status of these genes in HCC and non-tumor liver tissues using sequencing and MSP, and evaluated their use, individually and in combination, as a diagnostic tool for the detection of early HCC.…”

CpG methylation of MGMT and hMLH1 promoter in hepatocellular carcinoma associated with hepatitis viral infection

“…Esteller et al (2001) have demonstrated the reduced expression of tumour suppressor genes such as p16, MGMT, and hMLH1 by promoter hypermethylation in several human neoplasias, and have suggested that this epigenetic change might be an early event in the pathogenesis of several human tumours. Recently, the correlation of p16 promoter hypermethylation with chronic hepatitis and cirrhosis associated with HBV or HCV infection has been reported (Kaneto et al, 2001). …”