Abstract. The accurate detection of the presence of lymph node metastases (LNM) of gastric cancers (GCs) is useful for the implementation of necessary and sufficient treatment, but current methods of detection are unsatisfactory. In the present study, we focused on DNA methylation markers since they have several advantages, including biological and chemical stability and informativeness even in the presence of contaminating cells. Using three metastatic lymph nodes and three primary GCs without LNM, methylation bead array analyses were performed, which enabled the interrogation of 485,577 CpG sites. A total of 31 CpG sites that were hypermethylated in the metastatic lymph nodes, compared with the GCs without LNM, were isolated. Using primary GCs with and without LNM (28 GCs with LNM and 10 without), their methylation levels were measured using quantitative PCR following treatment with sodium bisulfite or a methylationsensitive restriction enzyme. Of the genomic regions around the 31 CpG sites, 10 regions demonstrated higher methylation levels in the GCs with LNM compared with the GCs without LNM (P<0.05). Finally, the hypermethylation of the 10 regions was validated using another set of samples (129 GCs with LNM and 20 without). Hypermethylation of the region around the cg06436185 CpG site predicted the presence of LNM at a sensitivity of 43% and specificity of 85%. Additionally, the hypermethylation of the region was associated with a poor survival rate among GC patients with LNM. The results of the present study indicated that the methylation status of the region was a promising candidate marker to detect the presence of LNM of GCs and may reflect the malignant potential of GCs.
IntroductionGastric cancer (GC) is one of the most prevalent malignancies worldwide and remains a leading cause of cancer-related mortality (1,2). Since the presence of lymph node metastases (LNM) is associated with a significantly poorer prognosis of GC patients (3-5), radical resection with free-margin gastrectomy and extended lymphadenectomy are performed for patients with advanced GC to eradicate LNM (6). Such an aggressive resection of the lymph nodes is associated with higher patient morbidity and/or mortality rates (7-9). Alternatively, the absence of LNM allows for minimally invasive surgery, which provides an improved quality of life following treatment. Therefore, the accurate detection of LNM is useful for the implementation of necessary and sufficient treatment.To detect the presence of LNM, much effort has been made in the fields of imaging and molecular markers. Imaging modalities, including computed tomography (CT), endoscopic ultrasonography (EUS) and 18 F-fluorodeoxyglucose positron emission tomography (FDG-PET) are used in clinical practice. However, the sensitivities of these modalities are 77.2, 82.8 and 71%, respectively, and the specificities are 78.3, 74.2 and 74%, respectively (10-13). Moreover, these imaging modalities are almost powerless to detect micrometastases (14,15). With regard to molecular markers, an...