CDCA8 and BIRC5 are downregulated in DHL cells after treatment with OR-2100. GO analysis of the genes that were downregulated by OR2100 monotherapy (A) or AZA monotherapy (B). (C) Volcano plot showing the DEGs between the Ctr group and OR-2100–treated samples. |log2 fold change| > 2; adjusted P <.05. Red indicates the upregulated genes, and blue indicates the downregulated genes. CDCA8, BIRC5, INCENP, and AURKB were identified as significantly downregulated genes. (D) Histograms of the percentage CpG methylation of RC cells and OCI-Ly18 cells after treatment with vehicle (Ctr) or OR2100. (E) The relative expression of CDCA8 and BIRC5 was determined by real-time PCR after treatment with the vehicle (Ctr), DAC, or OR-2100. The relative expression is shown as the fold change relative to the untreated cells after normalization to the expression of β-actin. (F) The expression of DNMT-1, β-tubulin, borealin (CDCA8), and survivin (BIRC5) proteins in RC cells and OCI-Ly18 cells that were treated with vehicle (Ctr) or the HMAs AZA, DAC, or OR-2100 was determined by western blotting. The representative images show that DAC and OR-2100 reduced the expression of the DNMT-1, borealin, and survivin proteins when compared with AZA. (G) STRING network analysis of the functional association between CDCA8 and BIRC5. INCENP and AURKB, parts of the CPC together with CDCA8 and BIRC5, show mutual protein association. Red, positive regulation of mitotic cytokinesis; green, mitotic spindle assembly checkpoint signaling; blue, SGO1. (H) A schematic image of the CPC. Ctr, control; DNMT-1 DNA methyltransferase inhibitor 1. Created with BioRender.com.