Objective:To explore the causal relationship between one-carbon metabolism related vitamin B levels and the occurrence of esophageal cancer(EC)using mendelian randomization(MR)analysis.Methods:One-carbon metabolism related vitamin B levels in the serum and EC were considered as the exposure and outcome variables,respectively.Single nucleotide polymorphisms(SNPs)significantly associated with the content of vitamin B6,B9,and B12 were selected as instrumental variables(IVs)to perform MR analysis of the causal association.Heterogeneity analysis was conducted using Cochran's Q test.Horizontal pleiotropy was tested using MR-Egger regression and MR-PRESSO tests.The robustness of the results was examined using the leave-one-out method.Results:A total of 24,18,and 25 SNPs were included as IVs for vitamin B6,B9,and B12,respectively.The inverse variance weighted odds ratio(OR)values were 1.001,0.972,and 1.000(all P>0.017).Cochran's Q test indicated no heterogeneity(all P>0.05).Neither MR-Egger regression nor MR-PRESSO tests found horizontal pleiotropy(all P>0.05).Leave-one-out sensitivity analysis suggested that the results were robust.Conclusion:There is no direct causal relationship between one-carbon metabolism related vitamin B levels and the occurrence of EC.