Reference gene selection for real-time RT-PCR in eight kinds of rat regenerating hepatic cells.

Liver regeneration (LR) is a process during which the liver recovers its mass and function after damage due to various causes such as partial hepatectomy (PH). It involves a sequence of well-orchestrated changes in physiological and biochemical activities, especially in the gene expression profile in a variety of liver cells. In order to produce reliable gene expression of target genes in eight kinds of rat hepatic cells during LR, the determination of internal control housekeeping genes (HKGs) is required. Eight kinds of hepatic cells were first isolated from liver tissue with high purity and activity. Then quantitative real-time reverse transcription (RT)-PCR was applied to detect expression changes of six commonly used HKGs (18SrRNA, B2M, ACTB, UBC, GAPDH, and HK1) in eight types of hepatic cells isolated from regenerating liver at 0, 2, 6, 12, 24, 30, 36, 72, 120, and 168 h after PH. The amplification of the HKGs was statistically analyzed by using geNorm algorithm. Using this method, 18SrRNA-UBC, ACTB-HK1, ACTB-GADPH, B2M-ACTB, 18SrRNA-UBC, B2M-UBC, B2M-ACTB, and B2M-UBC were found to be the two most stable reference genes for rat regenerating hepatocytes, hepatic stellate cells, Kupffer cells, biliary epithelial cells, sinusoidal endothelial cells, pit cells, dendritic cells, and oval cells, respectively, regardless of the stages of LR. In conclusion, this study has laid a good foundation for investigating gene expression of target genes in different types of hepatic cells during LR.