Abstract: Objective：To investigate the methylation status of DLC-1 in retinoblastoma (RB) tissue and its correlation with clinical parameters. Methods：Thirty-seven paraffin embedded RB tissue samples, 4 normal eye tissue samples and 2 retinoblastoma cell line (Y79 and WERI-Rb1) samples were collected, and their methylation status of DLC-1 were quantitatively detected with the methylation-sensitive high resolution melting (MS-HRM) method. Then, the bisulfite genomic sequencing was used to validate the methylation status of DLC-1 gene. In the meanwhile, the expression of  DLC-1 mRNA in Y79 and WERI-Rb1 cell lines were determined by RT-PCR. Results：Aberrant DLC-1 methylation was detected in 13 (35.1%) of 37 RB samples, but was not found in normal eye tissues. DLC-1 mRNA was detected in Y79 and WERI-Rb1 cell lines. The DLC-1 methylation was not correlated with tumor differentiation, other accompanied cancers and family history. However, the methylation rate of DLC-1 promoter in unilateral RB patients was significantly higher than that in bilateral patients (P＜0.05). Conclusion：Aberrant methylation of DLC-1 promoter exists in RB patients, and may play a role in the pathogenesis of RB.