Myc-associated zinc finger protein drives colorectal cancer metastasis through activating ubiquitin like with ring finger protein one

Figure 3 Myc-associated zinc finger protein promotes esophageal cancer-related gene 4 DNA methylation via ubiquitin-like with PHD and RING finger domains 1 activation. A and B: Western blot and quantitative reverse transcription polymerase chain reaction (PCR) showed ubiquitin-like with PHD and RING finger domains 1 (UHRF1) and esophageal cancer-related gene 4 (ECRG4) level in colorectal cancer cell lines; C: Dual luciferase reporter assay showed binding of Myc-associated zinc finger protein to UHRF1 promoter; D and E: Western blot and quantitative reverse transcription PCR demonstrated UHRF1 knockdown efficiency and its effect on ECRG4 expression; F: Chromatin immunoprecipitation-quantitative PCR showed that UHRF1 was enriched at the ECRG4 promoter in colorectal cancer cells; G: Methylation-specific PCR showed methylation and unmethylation status of the ECRG4 promoter after UHRF1 knockdown. Data are presented as the mean ± SD. The dashes represent the comparisons between groups. The experiments were repeated at least three times independently. MAZ: Myc-associated zinc finger protein; UHRF1: Ubiquitin-like with PHD and RING finger domains 1; ECRG4: Esophageal cancer-related gene 4; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; sh: Short hairpin; NC: Negative control; oe: Overexpression. ^aP < 0.05, ^bP < 0.01, and ^cP < 0.0001.