Cooperative regulation of CYP2C12 gene expression by STAT5 and liver-specific factors in female rats.

The purpose of this study was to clarify the mechanism(s) responsible for the growth hormone (GH)-induced expression of the CYP2C12 gene. To identify a functional GH-responsive element (GHRE) in vivo, we performed the direct injection of promoter-luciferase chimeric genes into female rat livers. The results showed that the luciferase activity was decreased to approximately 20% by the deletion of the sequence between nucleotides -4213 and -4161. Within this region, two copies of a possible GHRE were present. The sequence of the GHRE was overlapped with that of an interferon-gamma-activated sequence, known to be recognized by the signal transducer and activator of transcription (STAT) proteins. In fact, a supershift assay showed that STAT5 was capable of binding to the core sequence of the GHRE. Furthermore, a luciferase assay with reporter plasmids, Delta-4161/-3781, mutated hepatocyte nuclear factor-4 (HNF-4), and mutated HNF-6, revealed that the GH-stimulated expression of the CYP2C12 gene was regulated cooperatively by STAT5, HNF-4, HNF-6, and the factor(s) that binds to the elements, 2C12-I (-4095 to -4074) and 2C12-II (-4072 to -4045). The cooperative regulation by STAT5 and the liver-enriched transcription factors account for the GH-dependent and the liver-specific expression of the CYP2C12 gene in female rats.