Abstract
Hypermethylation of specific gene promoters is an important mechanism of carcinogenesis. A high frequency of promoter methylation of PAX1 and ZNF582 genes has been detected in cervical cancer. In the present study, we investigated the methylation status of PAX1 and ZNF582 genes in esophageal squamous cell carcinoma (ESCC) tissues. Tumor and paracancerous tissues were obtained from 14 ESCC patients. Genomic DNA was extracted from both tumor and paracancerous tissues, and the concentration of DNA were determined. DNA methylation analysis of PAX1 and ZNF582 genes was carried out using quantitative methylation-specific PCR. To assess the diagnostic performance of the two methylated genes for cancer detection, receiver operating characteristic (ROC) curves were generated. Sensitivities and specificities were tested at cut-offs obtained from the ROC curves. The methylation levels of both PAX1 and ZNF582 genes were significantly higher in tumor tissues compared to non-tumor paracancerous tissues. The methylation rates of PAX1 and ZNF582 in ESCC tumor and paracancerous tissues were 100% and 21.4% (p = 0.006), 85.7% and 0% (p < 0.001), respectively. The sensitivities and specificities of PAX1 and ZNF582 methylation for the detection of cancer were 100% and 85.7%, and 78.6% and 100%, respectively. The DNA methylation levels and frequencies of PAX1 and ZNF582 genes were markedly higher in ESCC tumor tissues compared to those in paracancerous tissues. Moreover, the conclusions were verified by using The Cancer Genome Atlas (TCGA) datasets. DNA methylation status of these two genes showed a relatively good sensitivity and specificity for the detection of ESCC tumors. This data suggests that DNA methylation testing holds a great promise for ESCC screening and warrants further prospective population-based studies.
Highlights
Esophageal cancer (EC) is one of the most aggressive cancers
A total of 14 subjects with esophageal squamous cell carcinoma (ESCC) were recruited from Xiangya Hospital located in Changsha, China between May and November 2015
Previous studies have shown that the methylation status of Paired boxed gene 1 (PAX1) and zinc finger protein 582 (ZNF582) genes is a useful testing to distinguish tumor and non-tumor tissues in cervical and oral squamous cell carcinoma [19,20]
Summary
This study was approved by the Institutional Review Board of Department of Clinical. Tumor and paracancerous tissues were obtained from 14 patients with ESCC, who had surgery to resect the tumor between May and November 2015. Inclusion criteria were as follows: (a) patients with ESCC who needed surgical resection; and (b) cases with a sufficient amount of residual tumor and paracancerous tissues for DNA methylation analysis. Histopathological examination was performed to characterize the tumor tissues, and three to five tumor samples were taken from each subject. Paracancerous tissues were taken from surgically dissected tissues approximately 2 cm away from the defined tumor area without tumor invasion by histopathology. The dissected tumor and paracancerous tissues were further evaluated by histological examination. Tumor samples were considered pure and acceptable for inclusion in this study
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