细胞周期调控基因及Bcl-2、c-Jun基因在人肝细胞脂肪变性过程中的表达变化

摘要

Objective To investigate the expression changes of cell cycle regulatory genes [Cyclin-dependent kinase2 (CDK2),Cyclin-dependent kinase4 (CDK4),and CyclinD1],B-cell lymphoma/leukemia-2 (Bcl-2),and c-Jun in the process of human hepatic steatosis.Methods L02 liver cell line was incubated with mixture of oleic acid and palmitic acid by the ratio of 2 ∶ 1 (OP21) for 0,5,10,and 20 h to build the models of human hepatocyte steatosis.Oil Red O staining was performed to evaluate the fat content of the L02 liver cells.The mRNA expression levels of CDK2,CDK4,CyclinD1,Bcl-2,and c-Jun in the L02 cells at different time points were detected by real-time PCR.Results More and more orange droplets were accumulated in the L02 cells with the time goes on,and the liver cells were dyed orange at 20 h.The mRNA expression level of CDK2 increased gradually,reached the peak at 5 h (P ＜ 0.05),and then decreased gradually;the mRNA expression level of CDK4 increased gradually,reached the peak at 10 h (P ＞ 0.05),then decreased gradually,and reached the lowest point at 20 h (P ＞ 0.05);no statistically significant difference was found in the expression levels of Cyclin D1 at different time points during the process (all P ＞ 0.05);the mRNA expression level of Bcl-2 increased over time,reached the peak at 10 h (P ＜0.05),and then decreased;the mRNA expression level of c-Jun was higher at 5,10,and 20 h than that at 0 h (all P ＜ 0.05),but there were no regular changes.Conclusion During the process of hepatic steatosis,the relative mRNA levels of the cancer-related genes (CDK2,CDK4,Bcl-2,and c-Jun) first increase and then decrease,while the expression of Cyclin D1 does not change significantly.%目的 观察细胞周期调控基因及Bcl-2、c-Jun基因在人肝细胞L02脂肪变性过程中的表达变化.方法 以油酸和软脂酸(2∶1)混合液处理L02细胞0、5、10、20h,建立人肝细胞脂肪变性模型.用油红O染色检测L02细胞内脂质含量;用荧光定量PCR技术检测L02细胞脂肪变性过程中CDK2、CDK4、CyclinD1、Bcl-2、c-Jun mRNA表达.结果 随造模时间延长,L02细胞内橘红色脂滴聚集越来越多,至造模20h,肝细胞染成橘红色.L02细胞内CDK2 mRNA相对表达量逐渐升高,造模5h达最高(P均＜0.05),随后逐渐下降;CDK4 mRNA相对表达量逐渐升高,造模10 h达最高(P均＞0.05),随后逐渐下降,造模20h最低(P均＞0.05);不同时间点Cyclin D1 mRNA相对表达量比较差异无统计学意义(P均＞0.05);Bcl-2 mRNA相对表达量随造模时间延长而升高,造模10 h达最高(P＜0.05),随后降低;造模5、10、20h的c-JunmRNA相对表达量均较造模0h高(P均＜0.05),但无变化规律.结论 人肝细胞L02脂肪变性过程中癌症相关基因CDK2、CDK4、Bcl-2、c-Jun表达先升高后降低,Cyclin D1表达变化不明显.