KDM4A调控肝癌细胞增殖机制  

Mechanism of KDM4A in the Regulation of Hepatocellular Carcinoma Cell Proliferation

作　　者：孙黎[1] 戴宇翃 邹曼 邱红 SUN Li;DAI Yuhong;ZOU Man;QIU Hong(Department of Oncology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology, Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属同济医院肿瘤科

出　　处：《医药导报》2019年第7期856-859,共4页Herald of Medicine

基　　金：国家自然科学基金资助项目(81372664)

摘　　要：目的探讨组蛋白赖氨酸去甲基化酶KDM4A(JMJD2A)调控肝癌细胞增殖的作用及机制。方法293T细胞中包装慢病毒(Lenti-shKDM4A#1,Lenti-shKDM4A#2,Lenti-shctr,Lenti-HA-Myr-AKT,Lenti-HA-vector)。在肝癌细胞系Hep3B中建立KDM4A稳定低表达细胞系及对照细胞系。克隆形成实验检测KDM4A低表达之后细胞增殖状况变化。WesternBlot分析KDM4A对AKT通路激活的影响;通过在KDM4A低表达细胞中稳定表达Myr-AKT,观察AKT信号"Rescue"后细胞生长状态,确定KDM4A依赖于Akt调节肝癌细胞增殖。结果两个KDM4A低表达Hep3B细胞系shKDM4A#1(P<0.01)和shKDM4A#2(P<0.001)克隆形成数目较对照组明显减少。KDM4A低表达后AKT磷酸化水平降低,AKT下游信号P-GSK-3b(ser9)蛋白表达水平显著下降。而在shKDM4A低表达细胞中表达Myr-AKT能够逆转KDM4A对Hep3B细胞增殖的影响。结论KDM4A能够通过激活AKT调控肝癌细胞增殖。Objective To investigate the effect and mechanism of histone lysine demethylase KDM4 A( JMJD2 A) in the regulation of hepatocellular carcinoma( HCC) cell proliferation. Methods The lentiviral particles,including Lenti-shKDM4 A#1,Lenti-shKDM4 A # 2,Lenti-shctr,Lenti-HA-Myr-AKT and Lenti-HA-vector, were packaged in 293 T cells. The KDM4 A knockdown model and control cells were established in Hep3 B cell line. Colony formation assay was used to observe the cell proliferation in sh KDM4 A and control cells.AKT activation was detected by Western Blot.The Myr-AKT was stably over-expressed in KDM4 A knockdown cells,then the cell growth condition was observed to confirm the role of KDM4 A in cell proliferation depending on AKT activation. Results Two KDM4 A knockdown cell lines,sh KDM4 A#1( P< 0.01) and sh KDM4 A#2( P<0.001),formed significantly less clones than the control group. The phosphorylation level of AKT dramatically decreased after KDM4 A knockdown,and AKT downstream signal protein P-GSK-3 b( ser9) expression level reduced correspondingly.Furthermore,overexpression of Myr-AKT in sh KDM4 A Hep3 B cells could reverse the effect of KDM4 A on cell proliferation.Conclusion KDM4 A regulates the HCC cell proliferation depending on AKT activation.

关 键 词：KDM4A(JMJD2A) 肝癌 AKT GSK-3b 

分 类 号：R979.1[医药卫生—药品] R965[医药卫生—药学]