Aims: CpG island methylator phenotype (CIMP) involves the targeting of multiple genes by promoter hypermethylation, and the cell-cycle regulatory proteins often change in human neoplasms. To gain insight into the role of epigenetic aberration of cell-cycle regulator genes in oesophagus squamous cell carcinoma (ESCC), the authors determined the hypermethylation profile in ESCC. Methods: The promoter methylation status of nine cell-cycle regulator genes was examined in 50 ESCC, 50 dysplastic tissues and 50 normal epithelial tissues by methylation-specific PCR. Results: The frequency of promoter methylation was 52% for p14, 44% for p15, 50% for p16, 56% for p21, 38% for p27, 8% for p53, 42% for p57, 36% for p73, and 44% for RB1 of 50 ESCC. CIMP+ was detected in 54% (27/50) of ESCC and 8% (4/50) of dysplastic tissues; no CIMP+ was present in normal epithelial tissues (p<0.001). The results show that promoter methylation of p14, p15, p16, p21, p27, p57 and p73 was far more common in ESCC samples with CIMP+ than those with CIMP−. A significant difference between CIMP status and TNM stage and metastasis was found in ESCC (p<0.05). Patients with ESCC with CIMP+ had poorer 4-year survival than those with CIMP−. Conclusions: The results suggest that CIMP of a subset of cell-cycle regulatory genes has an important role in the pathogenesis and progression of ESCC.

Background: Epigenetic alterations of tumor-associated genes contribute to the pathogenesis of virtually all cancer types. We evaluated the methylation status of the interleukin-10 (IL-10) gene promoter and assessed its association with IL-10 mRNA expression and clinical prognosis in hepatocellular carcinoma (HCC) patients. Methods: Methylation-specific polymerase chain reaction (MSP) and real-time polymerase chain reaction (PCR) were used to define the methylation index (MI) of the IL-10 gene and quantify IL-10 mRNA expression in 120 HCC samples and paired non-tumor tissues. Results: Mean MI was 0.47 in HCC specimens and 0.59 in non-tumor controls, and was associated with metastasis classification and serum α-fetoprotein (AFP) levels. IL-10 mRNA levels [mean –∆∆Ct of 1.678 in HCC cases with hypomethylation (∆MI ≤0) and –0.18 in HCC cases with hypermethylation (∆MI >0)] also correlated with metastasis classification and serum AFP. An association was detected between IL-10 mRNA and its gene’s MI in HCC. Also, an association was found between IL-10 hypomethylation, but not IL-10 mRNA expression and reduced postoperative HCC survival. Conclusions: These results indicate that IL-10 promoter hypomethylation is associated with increased IL-10 mRNA levels and indicative of poor survival in HCC.

Esophageal cancer has high incidence rate in China. Neoadjuvant chemoradiotherapy (nCRT) has become the standard treatment for esophageal squamous cell carcinoma (ESCC). However, there are few reliable epigenetic parameters for patients with ESCC undergoing neoadjuvant therapy. Genomic extract from tumor tissue was amplified and sequenced using the Illumina HiSeq4000 to quantify genes associated methylation or hydromethylation in 12 patients with ESCC undergoing nCRT. The genome‑wide hydroxymethylation were analyzed by methylated and hydroxymethylated DNA immunoprecipitation sequencing by MACS2 software and UCSC RefSeq database. Abnormal DNA methylation was statistically different between nCRT‑well (showed a pathological complete response to nCRT) and nCRT‑poor (showed incomplete pathological response to nCRT) patients. Levels of ten‑eleven translocation 1, 2 and 3 mRNA and protein were higher in tumor tissue in nCRT‑well group patients than in nCRT‑poor group patients. Illumina HiSeq 4000 sequencing identified 2925 hypo‑differentially hydroxymethylated region (DhMRs) and 292 hyper‑DhMRs in promoter between nCRT‑well and nCRT‑poor patients. Biological processes associated with hyper‑DhMRs included ‘snRNA processing’, ‘hormone‑mediated signaling pathway’ and ‘cellular response’. Metabolic processes were associated with hypo‑DhMRs. These data may explain the functional response to nCRT in patients with abnormal promoter of methylation gene‑associated mRNA expression. The present results implied that hyper‑DhMRs and hypo‑DhMRs affect molecular pathways, such as hippo and Notch signaling pathways, highlighting epigenetic modifications associated with clinical response to nCRT in patients with esophageal cancer.

Background: Pro-inflammatory cytokine production, such as CD74 and macrophage migration inhibitory factor (MIF) could play an important role in liver cancer following chronic hepatitis B virus (HBV) infection. Methods: The level of CD74 was evaluated by using a capture enzyme-linked immunosorbent assay (ELISA) kit. The level of MIF protein was determined using Human Magnetic Luminex^® Assays on 60 HBV-related hepatocellular carcinoma (HCC) patients. The level of HBV copies was determined by using real-time polymerase chain reaction (RT-PCR). Furthermore, α-fetoprotein (AFP), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were determined by the cobas 6000 analyzer. Results: The mean concentration of CD74 was elevated significantly in the tumor tissues (10.24 ng/mg) when it was compared with non-tumor tissues (3.86 ng/mg). Meanwhile, the level of MIF was 63.49 ng/mg in the tumor tissues, and 45.47 ng/mg in non-tumor samples. We found that elevated CD74 protein expression was associated with an elevated serum AFP level and the AST/ALT ratio; however, an elevated MIF protein level was associated with a lymph node metastasis, serum AFP level and AST/ALT ratio. Only a high level of MIF protein was associated with a short-term survival rate of just months in the HBV-related HCC patients. The elevated expression of MIF protein was associated with HBV ≥10⁵ (copies/mL) in liver cancer. The elevated level of MIF protein was associated with the elevated level of CD74 protein in HBV-associated HCC progression. Conclusions: These results suggest that the elevated level of CD74 and MIF cytokines could play a significant role in HBV-related HCC.