Objective To research the upstream regulation mechanism of miR-133 b in esophageal squamous cell carcinoma.
Methods The upstream transcription factors involved in the regulation of miR-133 b expression were predicted by bioinformatics analysis. The effects of IRF4 overexpression plasmid on the expression of IRF4 and miR-133 b were verified by qRT-PCR and Western blotting. The mechanisms of IRF4 regulating miR-133 b expression were verified by chromatin immunoprecipitation(ChIP) and dual-luciferase reporter assay.
Results ConTra V3 predicted that the transcription factor IRF4 could bind to miR-133 b. The transfection of IRF4 overexpression plasmid could significantly up-regulate the expression of IRF4 at mRNA(
t=-18.950,
P=0.010) and protein(
t=-4.061,
P=0.042) levels, and promote the expression of miR-133 b(
t=-8.681,
P=0.005). ChIP( primer 1:
t=-17.391,
P=0.003; primer 2:
t=-14.421,
P=0.004) and dual-luciferase reporter assay(
t=-22.112,
P=0.010) confirmed that the transcription factor IRF4 could directly target miR-133 b.
Conclusion IRF4 can directly target miR-133 b, which is helpful to construct the signal regulatory network of miR-133 b and provide a new target for the diagnosis and treatment of esophageal squamous cell carcinoma.