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. 2014 Nov;42(11):1791–1795. doi: 10.1124/dmd.114.060145

Fig. 3.

Fig. 3.

Recombinant pre-mir-27b is effective in the modulation of CYP3A4 expression. (A) qPCR analyses revealed that recombinant tRNA/mir-27b significantly (*P < 0.001, one-way analysis of variance) reduced CYP3A4 mRNA expression levels in LS-180 cells compared with the tRNA scaffold. GAPDH was used as an internal control. (B) Western blot analyses showed that LS-180 cells transfected with tRNA/mir-27b had lower CYP3A4 protein expression levels than the cells transfected with control tRNA. GAPDH was used as a loading control. (C) CYP3A4 enzymatic activity, as measured by [MDZ]/[1′-HO-MDZ] metabolic ratio at various time points after exposure to MDZ (at 72 hours post-transfection), was significantly (*P < 0.001, two-way analysis of variance) lower in LS-180 cells treated with tRNA/mir-27b than the tRNA/MSA control. MDZ and metabolite concentrations were determined by the liquid chromatography–tandem mass spectrometry method. Values are mean ± S.D. of triplicate treatments with separate cultures. *P < 0.05 compared with the control tRNA/MSA treatment.