Inhibitory effect of CYP450 3A4 gene expression and function by vector expressed small hairpin interfering RNA in CHL-3A4 cells lines

Objective To investigate the inhibitory effect of CYP 3A4 gene expression and function in CHL-3A4 cells lines by vector expressed small hairpin interfering RNA (shRNA). Methods The shRNA expression vectors targeting CYP 3A4 gene (CYP3A4Ⅰ,CYP3A4Ⅱ,CYP3A4Ⅲ) were designed and constructed. The cells were transfected with shRNA expression vectors transiently, and the cells without shRNA transfection and with nonspecific shRNA transfection were used as controls. The inhibitory effect of RNA interfering was detected by Western blot analysis.The activity of nifedipine oxydase in CHL-3A4 was mea- sured by HPLC assay.Results CYP 3A4III shRNA expressing vector significantly reduced the protein expression levels (75%) of the CYP 3A4 gene and significantly inhibited the activity of nifedipine oxydase in CHL- 3A4 cells.Conclusion Vector-based RNA could suppress CYP 3A4 gene expression and function, and the use of RNA interfering interfering to inhibit CYP3A4 gene expression in mammalian cells was a promising new tool for the study of gene function.