拉帕替尼抑制HL60细胞增殖和促进细胞凋亡的机制  

Mechanism underlying inhibition of proliferation and promotion of apoptosis by lapatinib in HL60 cells

作　　者：刘路 刘北忠[1,2] 赵毅 陈敏 姚仕菲 李连文 肖春兰[1] 单志灵 徐婷[1] 淦柳根 钟梁[2] LIU Lu;LIU Beizhong;ZHAO Yi;CHEN Min;YAO Shifei;LI Lianwen;XIAO Chunlan;SHAN Zhiling;XU Ting;GAN Liugen;ZHONG Liang(Central Laboratory, Yongchuan Hospital, Chongqing Medical University, Chongqing 402160;Ministry-of-Education Key Laboratory of Laboratory Medical Diagnostics, Faculty of Laboratory Medical, Chongqing Medical University, Chongqing 400016, China)

机构地区：[1]重庆医科大学附属永川医院中心实验室,重庆402160 [2]重庆医科大学临床检验诊断学教育部重点实验室重庆市重点实验室,重庆400016

出　　处：《细胞与分子免疫学杂志》2017年第10期1341-1347,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81171658);重庆市自然科学基金(2011BA5037)

摘　　要：目的探讨拉帕替尼对急性髓细胞白血病HL60细胞增殖和凋亡的影响及其相关分子机制。方法用(5、10、15)μmol/L的拉帕替尼处理HL60细胞24 h,光学显微镜下观察细胞形态变化,CCK-8法检测细胞活力,集落形成实验检测细胞增殖能力,异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双标记法结合流式细胞术检测细胞凋亡,Wright改良染色(LIU染色)和Hoechst33342荧光染色观察细胞核形态;Western blot法检测Bax、Bcl2、胱天蛋白酶3(caspase-3)、caspase-9、裂解型caspase-3(c-caspase-3)、裂解型caspase-9(c-caspase-9)、裂解型多腺苷二磷酸核糖聚合酶(c-PARP)、蛋白磷酸酶2A细胞增殖调节抑制因子(CIP2A)、c-MYC、AKT和磷酸化AKT(p-AKT)蛋白水平。结果与对照组相比,拉帕替尼能以剂量依赖的方式抑制HL60细胞的增殖,促进细胞凋亡,诱导细胞产生核碎裂、染色质凝聚。下调Bcl2蛋白的表达,上调Bax、c-caspase-3、c-caspase-9和c-PARP蛋白水平,下调CIP2A、p-AKT和c-MYC蛋白水平。结论拉帕替尼能够抑制HL60细胞增殖并促进细胞凋亡,这一作用可能与下调CIP2A/AKT/c-MYC信号通路有关。Objective To investigate the effect of lapatinib on cell proliferation and apoptosis in acute myeloid leukemia HL60 cells in vitro and the related molecular mechanisms. Methods The HL60 cells were treated with 5,10,15 μmol/L lapatinib for 24 hours,and then morphological changes of the cells were observed under optical microscope. CCK-8 assay was used to assess the cell viability. Colony formation assay was performed to detect the cel proliferation ability. Cel apoptosis labeled by annexinⅤ-FITC/PI were analyzed by flow cytometry. Wright modified LIU＇s staining and Hoechst33342 fluorescent staining were used to observe the morphology of the nucleus. Western blotting was utilized to detect the expressions of Bax,Bcl2,caspase-3, caspase-9, cleaved caspase-3, cleaved caspase-9, cleaved poly-ADP-ribose polymerase（ cleaved PARP）,cell proliferation regulating inhibitor of protein phosphatase 2 A（CIP2 A）,c-MYC,AKT and p-AKT. Results Compared with the control group,lapatinib inhibited cell proliferation and promoted apoptosis,induced nuclear fragmentation,chromatin condensation of HL60 cells in a dose-dependent manner. Meanwhile,it down-regulated the expression of Bcl2,up-regulated the levels of Bax,cleaved caspase-3,cleaved caspase-9 and cleaved PARP,and decreased the levels of CIP2 A,p-AKT and c-MYC. Conclusion Lapatinib could inhibit cell proliferation and promote apoptosis in HL60 cells by inhibiting the CIP2 A/AKT/c-MYC signal pathway.

关 键 词：拉帕替尼 HL60细胞 细胞增殖 细胞凋亡 

分 类 号：Q279[生物学—细胞生物学] R392.5[医药卫生—免疫学]