Induction of hormone-sensitive lipase/cholesteryl esterase gene expression by C/EBPα independently of the PKA pathway in the adrenocortical Y-1 cells

• Published in: Steroids 2015-12, Vol.104, p.118-121
• Main Authors: Czajkowski, M.T., Holysz, M., Trzeciak, W.H.
• Format: Article
• Language: English
• Subjects: Adrenal Cortex - cytology; Adrenal Cortex - metabolism; Animals; C/EBPα; CCAAT-Enhancer-Binding Protein-alpha - metabolism; Cell Line; Cyclic AMP-Dependent Protein Kinases - metabolism; Gene Expression Profiling; Lipase - genetics; Lipase - metabolism; LIPE induction; Mice; PKA; Real-Time Polymerase Chain Reaction; Sterol Esterase - genetics; Sterol Esterase - metabolism; Y-1 cells
• ISSN: 0039-128X; 1878-5867
• DOI: 10.1016/j.steroids.2015.09.003

•C/EBPα induces LIPE expression encoding HSL one of the key enzymes of steroidogenesis.•C/EBPα effect on LIPE expression is independent of the stimulation of the PKA signaling pathway.•C/EBPα binds to a response element located in the −60bp fragment of LIPE promotor A. The effect of C/EBPα on the expression of LIPE gene encoding hormone-sensitive lipase/cholesteryl esterase (HSL) was investigated in Y-1 CCL79 cells. It was found that transfection of these cells with the vector overexpressing C/EBPα increased both the level of LIPE transcript, measured by RT-qPCR, and the luminesce emitted by luciferase reporter gene fused to the −2150 fragment of LIPE promoter. Activation of adenylyl cyclase by forskolin resulted in 2.5-fold increase in the intensity of luminescence and over 3-fold increase in luminescence was observed when the cells were cotransfected with the vector overexpressing C/EBP. The incubation of C/EBP-cotransfected cells with forskolin caused over 6-fold increase in the intensity of luminescence, suggesting that the effects of C/EBPα and forskolin are additive. The analysis of sequence of the proximal LIPE promoter showed multiple binding sites for various transcription factors including C/EBPα site, which is located between nucleotides −46bp and −59bp. When the Y-1 cells were transfected with the recombinant vector containing −60bp fragment of LIPE promoter fused to the luciferase reporter gene and were cotransfected with the vector overexpressing C/EBPα, the luminescence increases about 9-fold indicating that C/EBPα stimulates the expression of LIPE by reacting with its response element. The results indicate that C/EBPα stimulates the expression of LIPE independently of the PKA pathway by binding to a response element situated within the −60bp fragment of LIPE promoter. This suggests that C/EBPα might be involved in the regulation of LIPE expression and thus cholesterol supply for steroid hormone synthesis.