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目的:阐明NDRG2(N-Myc downstream-regulated gene 2)在大鼠肝再生过程中对肝细胞周期的调控机制。方法:大鼠70%肝切除后收集0 h、8 h、24 h、48 h、72 h、7 d、10 d的再生肝组织,采用Real-time PCR和Western blot方法检测大鼠肝再生过程中NDRG2基因和蛋白的动态变化。流式细胞仪检测腺病毒介导的高表达NDRG2对大鼠正常肝细胞系(BRL)细胞周期的影响。Real-time PCR和Western blot方法检测高表达NDRG2对肝细胞周期调控的分子机制。结果:NDRG2基因和蛋白的表达水平在肝再生达到高峰时明显下降,在肝细胞进入分化期时显著上调;流式细胞仪检测显示BRL细胞中高表达NDRG2 48 h后,G0/G1期细胞百分比从对照组39.30±1.97上升至57.44±2.56,S期从37.66±1.73下降至13.27±2.01,差异有统计学意义(P<0.05)。对周期调控相关分子的检测显示高表达NDRG2对肝细胞周期的影响是通过上调p21,抑制Cyclin E实现的。结论:NDRG2通过影响细胞周期参与调控大鼠肝再生过程。
Objective: To elucidate the regulatory mechanism of N-Myc downstream-regulated gene 2 on hepatocyte cycle in rat liver regeneration. Methods: Liver tissues of 0, 8, 24, 48, 72, 7 and 10 days after hepatectomy were collected in rats. Real-time PCR and Western blot were used to detect the liver regeneration In NDRG2 gene and protein dynamics. Flow cytometry was used to detect the effect of adenovirus-mediated high expression of NDRG2 on the cell cycle of rat normal liver cell line (BRL). Real-time PCR and Western blot were used to detect the molecular mechanism of hepatocyte cycle regulation by highly expressed NDRG2. Results: The expression of NDRG2 gene and protein decreased significantly at the peak of liver regeneration and significantly increased when hepatocytes entered the differentiation stage. Flow cytometry showed that the percentage of cells in G0 / G1 phase increased from The control group increased from 39.30 ± 1.97 to 57.44 ± 2.56, while the S phase decreased from 37.66 ± 1.73 to 13.27 ± 2.01, with significant difference (P <0.05). Detection of the molecules involved in cycle regulation showed that the effect of high expression of NDRG2 on the cell cycle of hepatocytes was mediated by upregulation of p21 and inhibition of Cyclin E. Conclusion: NDRG2 regulates rat liver regeneration by affecting cell cycle.