Background: Vitamin D₃ has been known to have an anticancer effect, but the mechanisms underlying this is poorly explored. The present study aimed to investigate the antitumor role of vitamin D₃ on gastric cancer and mechanisms.
Methods: The Roche Elecsys platform was applied in retrospective studies to detect the role of 25-hydroxylvitamin D₃ in adenocarcinoma and colony formation assay was conducted to verify the effect of 1, 25-dihydroxyvitamin D₃ on the proliferation of gastric cancer cells. After the identification of hypermethylation of BMP3 CpG islands by bisulfite genomic sequencing (BGS), we further investigated the relationship of BMP3 expression and gastric carcinogenesis by Western blot analysis and gel electrophoresis mobility shift assay (EMSA). 
Results: Here we show that low concentration of 1, 25-dihydroxyvitamin D₃ links to cancerization and significantly inhibits proliferation of undifferentiated gastric cancer cell lines SGC-7901 and BGC-823. BMP3 promoter hypermethylation was highly correlated with gastric tumor. Moreover, BMP3 expression was regulated by its promoter methylation in gastric cells. The further exploration of the relationship between 1, 25-dihydroxyvitamin D₃ and BMP3 by EMSA results that 1, 25-dihydroxyvitamin D₃ stimulates BMP3 expression by the inhibition of BMP3 promoter methylation in gastric tumor cells. 
Conclusion: In combination with the data from clinical research, bioinformatics analysis and experimental verification, we propose that 1, 25-hydroxylvitamin D₃ affects gastric cancer progression by repressing BMP3 promoter methylation.
Keywords: BMP3, 1,25-dihydroxyvitamin D₃, gastric cancer, promoter methylation