Epigenetic modifications via DNA methylation and long non-coding RNAs (lncRNAs) have been identified in
bladder cancer (BC). However, DNA methylation of lncRNAs involved in BC has not been elucidated. Here,
DNA immunoprecipitation-sequencing (MeDIP-seq) and
RNA-sequencing (
RNA-seq) were carried out using eight paired
tumor and adjacent normal tissue samples from patients with BC. Differences in methylation patterns between
tumors and controls were compared and the percentage of differentially methylated genes, including
lncRNA genes, was calculated.
RNA-seq data were subjected to gene ontology (GO), Kyoto encyclopedia of genes, and genomes (KEGG) analysis. The association between DNA methylation modification and
lncRNA expression was determined by pairwise analysis of MeDIP-seq and
RNA-seq data. The most enriched motifs in the promoter region, as well as the methylated density in the 3 kb region surrounding super-enhancers of
lncRNA genes, were analyzed. A peak of 5mC methylation in the region 2 kb upstream of the transcription start site (TSS), with the lowest point in the TSS region, was observed. In total, 436 and 239 genes were identified to be hyper and hypomethylated, respectively, in BC tissue around the TSS region.
RNA-seq revealed differentially expressed lncRNAs between
tumor and normal tissues, many of which were
cancer-associated lncRNAs based on GO and KEGG pathway analysis. Combined MeDIP-seq and
RNA-seq analysis revealed that expression of 26 lncRNAs were candidates of 5mC controlled genes. The possible link between 5mC modification and differential lncRNAs may relate to enrichment of 5mC reads in the region surrounding super-enhancers of
lncRNA. Survival analysis indicated that the methylated
lncRNA, LINC00574, was associated with shorter overall survival time in patients with BC (HR = 1.7, p-value = 0.035). Taken together, these findings indicate that lncRNAs genes are under control of DNA methylation. Methylated
lncRNA genes, which are transcripted to LINC00574, may serve as
biomarkers for BC prognosis.