Combined Detection Of Plasma ZIC1,HOXD10 And RUNX3 Methylation For Early Diagnosis Of Gastric Cancer And Precancerous Lesions

ObjectiveWe try to evaluate the aberrant DNA methylation of multiple cancer-related genes in plasma in gastric cancer(GC)and precancerous lesions.We investigate combination detection of promoter methylation for ZIC1,HOXD10 and RUNX3 as non-invasive biomarker for early diagnosis of gastric caancer and the monitor of gastric carcinogenesis.MethodWe collect the plasma from healthy volunteers,patients with gastric intraepithelial neoplasias(GIN),gastric cancer(GC)and post surgery GC patients,and their clinicopathological parameters such as age,gender,tumor size,and TNM stage.Methylation-specific polymerase chain reaction(MSP)assays were used to measure DNA promoter methylation in plasma.The correlation between the positive rate of DNA promoter methylation and clinicopathological parameters of patients was analyzed.In addition,we investigate the clinical value of combined detection of ZIC1,HOXD10 and RUNX3 methylation for early diagnosis of gastric cancer and precancerous lesions.Result1)234 cases were enrolled which including 38 healthy volunteers(47.18±15.12 years old),55 GIN patients(59.71 ±10.62 years old),123 GC patients(40 early GC and 83 advanced GC)(63.7±9.38 years old)and 17 post surgery GC patients(59.75±9.75 years old).There were no significant differences in age and gender distribution among each groups(P>0.05).2)The rate of ZIC1,HOXD10 and RUNX3 methylation were 51.79%,33.92%and 37.50%in GIN group;67.50%,30.00%and 54.21%in early GC group;72.29%,54.21%and 40.96%in advanced GC group,respectively.Interesting,none of these genes was positively methylated in healthy controls.The methylation rate of ZIC1 in GC was higher than in GIN,while the methylation rate of HOXD10 in advanced GC higher than that in early GC(P<0.05).The methylation rates of HoxD10 in plasma in the GC group were decreased to 17.65%after surgery treatment(P<0.05).3)There were no statistically significant relationship between DNA promoter methylation and clinicopathological features such as age,gender,TNM stage,H.pylori infection status and serum level of CEA in GC patients(P>0.05).The methylated HoxD10 status in plasma was found to be associated with tumor invasion depth(P=0.023).4)Combined detection of ZIC1,HOXD10 and RUNX3 could elevate the sensitivity to 89.4%for diagnosis of gastric lesions(GIN and GC).In GC group,the sensitivity of ZIC1 was higher than HOXD10(46.3%)and RUNX3(42.3%).In GIN group,the specificity of HOXD10 was higher than ZIC1(69.1%)and RUNX3(78.7%).The sensitivity,specificity and AUC of combined detection of ZIC1 and HOXD10 was 83.1%,58.5%and 0.703,which were highest in all combined modes.ConclusionCombined detection of ZIC1,HOXD10 and RUNX3 promoter hypermethylation in plasma might be a promising strategy for early detection of GC and precancerous lesions in high risk population.