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Background  Colorectal cancer (CRC) arises as a consequence of genetic events such as gene mutation and epigenetic alteration. The aim of this study was to identify new hypermethylated candidate genes and methylation-based therapeutic targets using vincristine in CRC.    Methods  We analyzed the methylation status of 27,578 CpG sites spanning more than 14,000 genes in CRC tissues compared with adjacent normal tissues and normal colon tissues using Illumina bead chip array. Twenty-one hypermethylated genes and 18 CpG island methylator phenotype markers were selected as candidate genes. The methylation status of 39 genes was validated by quantitative methylation-specific polymerase chain reaction in CRC tissues, adjacent normal tissues, normal colon cells, and three CRC cell lines. Of these, 29 hypermethylated candidate genes were investigated using the demethylating effects of 5-aza-2′-deoxycytidine (5-aza-dC) and vincristine in CRC cells.    Results  Thirty-two out of 39 genes were hypermethylated in CRC tissues compared with adjacent normal tissues. Vincristine induced demethylation of methylated genes in CRC cells to the same extent as 5-aza-dC. The mRNA expression of  AKR1B1 ,  CHST10 ,  ELOVL4 ,  FLI1 ,  SOX5 ,  STK33 , and  ZNF304  was restored by treatment with 5-aza-dC and vincristine.    Conclusion  These results suggest that these novel hypermethylated genes  AKR1B1 ,  CHST10 ,  ELOVL4 ,  SOX5 ,  STK33 , and  ZNF304  may be potential methylation biomarkers and therapeutic targets of vincristine in CRC.

Identification of novel hypermethylated genes and demethylating effect of vincristine in colorectal cancer