C/EBPα-Per2信号通路对K562细胞相关基因的调控机制  

The mechanism research of C/EBPα-Per2 in K562 leukemia cells gene regulation

作　　者：孙成铭[1] 栾材富[1] 邵会媛[1] 高玉洁[1] 李少君[1] 马新衡[1] 刘日明[1] 李杰[1] 张守信[1] 刘鹏[1] 

机构地区：[1]青岛大学医学院附属烟台毓璜顶医院检验科中心,山东烟台264000

出　　处：《国际检验医学杂志》2014年第7期801-802,807,共3页International Journal of Laboratory Medicine

基　　金：山东省优秀中青年科学家科研奖励基金(BS2011YY064);烟台市科技发展计划项目(2011203)

摘　　要：目的探索Per2在CCAAT增强子结合蛋白α(C/EBPα)参与调控的慢性粒细胞白血病(CML)发生、发展中的作用。方法通过脂质体介导将真核表达载体pGenesil-3-SiPer2和空载体pGenesil-3-HK转染到pEGFP-C/EBPα-K562细胞,利用新霉素筛选稳定干扰Per2的pEGFP-C/EBPα-K562单克隆细胞株。利用RT-PCR和Western blot分别检测转染组、空载组、对照组细胞p53、c-myc、cyclinB1的mRNA及蛋白表达水平的改变。结果成功构建稳定干扰Per2表达的pEGFP-C/EBPα-K562细胞株。与对照组和空载组细胞相比,转染组pGenesil-3-SiPer2-K562细胞的p53mRNA及蛋白表达水平明显降低,差异有统计学意义(P<0.01),而cyclinB1和c-myc基因的mRNA及蛋白表达水平则显著升高,差异有统计学意义(P<0.01)。结论 Per2基因在C/EBPα参与调控的CML发生发展中起重要作用,该通路的发现对于研究CML的发生及调控机制具有重要意义。Objective To explore the role of Per2 in chronic myeloid leukemia（CML） with CCAAT/enhancer binding protein al- pha（C/EBPα）. Methods The pGenesil-3-SiPer2 and pGenesil-3-HK plasmid vector were transfected into pEGFP-C/EBPα-K562 cells, respectively. After screening culture by neomycin, cells stably reducing the expression of Per2 protein were established. RT- PCR and Western blot were used to detect the transcription and protein expression of p53,c-myc and cyclinB1 in the transfection group,the empty vector group and the control group. Results The pEGFP-C/EBPα-K562 cells with lower expression of Per2 were successfully established. Compared with cells in the empty vector group and the control group, the expressions of p53 mRNA and protein in the transfection group were significantly decreased（P〈0.01）, and the expressions of cyclinB1 and c-myc mRNA and pro- tein were significantly increased （P〈0.01） ,respectively. Conclusion This findings indicate that Per2 gene may play an important role in the development of CML with C /EBPα, which may have new value in investigating both the occurrence and regulatory mechanism of CML.

关 键 词：CCAAT增强子结合蛋白α 慢性粒细胞白血病 

分 类 号：R733.72[医药卫生—肿瘤]