LXR activation causes G1/S arrest through inhibiting SKP2 expression in MIN6 pancreatic beta cells

Liver X receptors (LXRs) are nuclear hormone receptors with central roles in lipid homeostasis. We previously showed that LXR activation induced aberrant lipid metabolism and G1 cell cycle arrest in pancreatic beta cells. In this study, we aimed to identify the molecular target of LXR causing G1 arrest. LXR activation was induced by its agonist, T0901317. A series of luciferase reporters of truncated Skp2 promoter were analyzed in MIN6 cells. mRNA and protein levels of SKP2 and P27 were detected. Flow cytometry assay was used to determine the cell cycle distribution. MTT assay was used to evaluate cell viability. LXR activation increased cell distribution in G1 phase and lipid accumulation. Since dominant-negative Srebp1c could clear the deposited lipid rather than recover the G1 arrest, we identified S-phase kinase-associated protein 2 (Skp2) as a potential target gene of LXR. In deed, LXR activation significantly inhibited Skp2 gene expression and protein amount. We also observed that the luciferase activity of Skp2 promoter was suppressed by T0901317 and the potential LXR regulatory site was narrowed down to a region of nt -289 to -38. Silencing Lxrα and Lxrβ rescued SKP2 protein level and recovered the cellular growth repressed by LXR activation. Moreover, SKP2 overabundance reduced P27 protein level by promoting its degradation, consequently overcame the G1 arrest caused by T0901317. Our findings demonstrate that transrepressing Skp2 expression by LXR activation resulted in defective SKP2-mediated P27 degradation and inhibitory cell growth in beta cells.