Potential diagnostic and prognostic values of detecting promoter hypermethylation in the serum of patients with gastric cancer

Abstract

While there is no reliable serum biomarker for the diagnosis and monitoring of patients with gastric cancer, we tested the potential diagnostic and prognostic values of detecting methylation changes in the serum of gastric cancer patients. DNA was extracted from the pretherapeutic serum of 60 patients with confirmed gastric adenocarcinoma and 22 age-matched noncancer controls. Promoter hypermethylation in 10 tumour-related genes (APC, E-cadherin, GSTP1, hMLH1, MGMT, p15, p16, SOCS1, TIMP3 and TGF-beta RII) was determined by quantitative methylation-specific PCR (MethyLight). Preferential methylation in the serum DNA of gastric cancer patients was noted in APC (17%), E-cadherin (13%), hMLH1 (41%) and TIMP3 (17%) genes. Moreover, patients with stages III/IV diseases tended to have higher concentrations of methylated APC (P = 0.08), TIMP3 (P = 0.005) and hMLH1 (P = 0.03) in the serum. In all, 33 cancers (55%) had methylation detected in the serum in at least one of these four markers, while three normal subjects had methylation detected in the serum (specificity 86%). The combined use of APC and E-cadherin methylation markers identified a subgroup of cancer patients with worse prognosis (median survival 3.3 vs 16.1 months, P = 0.006). These results suggest that the detection of DNA methylation in the serum may carry both diagnostic and therapeutic values in gastric cancer patients.

Figure 1

Frequency of detecting methylated DNA in the serum of gastric cancer patients and control. ^*Genes with differential methylation in cancer patients according to predefined criteria.

Figure 2

Number of genes methylated in the serum of gastric cancer patients and control.

Figure 3

Association between cancer staging and concentrations of methylated DNA. (A) APC (P=0.08); (B) E-cadherin (P=0.2); (C) TIMP3 (P=0.005) and (D) hMLH1 (P=0.03). The horizontal line indicated the median concentrations of the patients’ subgroups.

Figure 4

Kaplan–Meier analysis of the probability of overall survival in gastric cancer patients according to methylation status. (A) APC (P=0.20); (B) E-cadherin (P=0.09); (C) combined APC and E-cadherin (P=0.006).