Sprouty4（SPRY4）经常被报道为肿瘤抑制因子，因此在各种癌症中被下调。我们首次报告SPRY4在结直肠癌（CRC）中表观上调。在这项研究中，我们探讨了CRC细胞和患者样本中SPRY4的DNA甲基化和羟甲基化水平，并将这些发现与mRNA和蛋白质表达水平相关。使用联合亚硫酸盐限制分析方法评估了SPRY4启动子区域中的3个位点在CRC中的5mC水平。此外，还测量了CRC患者SPRY4内的羟甲基化水平。最后，从基因表达纲目和癌症基因组图谱等多个高通量数据库中提取了CRC患者的DNA甲基化和mRNA表达数据。综合体外和体内分析SPRY4启动子甲基化水平的结果清楚地表明，在CRC患者中，远端启动子区域经历了甲基化减少，并与增加的表达相关。此外，在SPRY4编码区域内发现了基因体羟甲基化的减少和基因体甲基化的增加，并与增加的表达相关。SPRY4在CRC中通过启动子甲基化减少和基因体超甲基化，表观上调，这值得进一步研究这种已确定的肿瘤抑制剂的非典型作用。

Sprouty4 (SPRY4) has been frequently reported as a tumor suppressor and is therefore downregulated in various cancers. For the first time, we report that SPRY4 is epigenetically upregulated in colorectal cancer (CRC). In this study, we explored DNA methylation and hydroxymethylation levels of SPRY4 in CRC cells and patient samples and correlated these findings with mRNA and protein expression levels. Three loci within the promoter region of SPRY4 were evaluated for 5mC levels in CRC using the combined bisulfite restriction analysis. In addition, hydroxymethylation levels within SPRY4 were measured in CRC patients. Lastly, DNA methylation and mRNA expression data were extracted from CRC patients in multiple high-throughput data repositories like Gene Expression Omnibus and The Cancer Genome Atlas. Combined in vitro and in silico analysis of promoter methylation levels of SPRY4 clearly demonstrates that the distal promoter region undergoes hypomethylation in CRC patients and is associated with increased expression. Moreover, a decrease in gene body hydroxymethylation and an increase in gene body methylation within the coding region of SPRY4 were found in CRC patients and correlated with increased expression. SPRY4 is epigenetically upregulated in CRC by promoter hypomethylation and hypermethylation within the gene body that warrants future investigation of atypical roles of this established tumor suppressor.