拉帕替尼通过激活p38MAPK信号通路促进NB4细胞凋亡  

Effects of Lapatinib on Cell Proliferation and Apoptosis in NB4 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] 

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：《中国细胞生物学学报》2017年第11期1390-1396,共7页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81171658);重庆市自然科学基金(批准号:2011BA5037)资助的课题~~

摘　　要：该文探讨了拉帕替尼对急性早幼粒细胞白血病NB4细胞增殖和凋亡的影响及相关分子机制。用p38MAPK抑制剂和不同浓度的拉帕替尼处理NB4细胞24 h,CCK-8(cell counting kit-8)实验检测细胞增殖,FITC-Annexin V/PI双染色法检测细胞凋亡,光学显微镜和Hoechst 33258染色观察细胞形态,Western blot检测Bcl-2(B cell leukemia-2)、Bax(Bcl-2 associated X protein)、caspase-3、PARP(poly-ADP-ribose polymerase)、PML-RARα(promyelocytic leukemia-retinoic acid receptor alpha)、p38MAPK(p38 mitongen-activated protein kinase)和p-p38MAPK(phosphorylated p38 mitongen-activated protein kinase)等蛋白质水平。结果显示,随着拉帕替尼药物浓度的增加,细胞增殖率显著降低,细胞凋亡数量明显增加,Hoechst 33258染色可见染色质浓缩、碎裂等凋亡现象。同时,拉帕替尼能降低Bcl-2和PML-RARα蛋白质水平,增加Bax、cleaved caspase-3、cleaved PARP和p-p38MAPK等蛋白质水平。用p38MAPK抑制剂预处理后,细胞增殖率升高,凋亡率降低,p-p38MAPK、Bax、cleaved caspase-3和cleaved PARP等蛋白质水平降低。该文结果提示,拉帕替尼能够抑制NB4细胞增殖并促进细胞凋亡,并且p38MAPK信号通路可能参与这些过程。This study was aimed to investigate the effect of lapatinib on cell proliferation and apoptosis in NB4 cells, and its related mechanisms. NB4 cells were treated with p38MAPK inhibitor and different concentrations of lapatinib for 24 h. The proliferation of NB4 cells was detected by CCK-8 （cell counting kit-8） assay. Apoptosis was determined by Annexin V/PI double binding assay and Hoechst 33258 fluorescent staining. Morphological changes were observed under optical microscope. The protein levels of Bcl-2 （B cell leukemia-2）, Bax （Bcl-2 associated X protein）, caspase-3, PARP （poly-ADP-ribose polymerase）, PML-RARa （promyelocytic leukemia-retinoic acid receptor alpha）, p38MAPK （p38 mitongen-activated protein kinase） and p-p38MAPK （phosphorylated p38 mitongen-activated protein kinase） were detected by Western blot. The results showed that lapatinib significantly inhibited proliferation and induced apoptosis of NB4 cells. Chromatin condensation and fragmentation were observed in lapatinib treated group. Lapatinib decreased the levels of Bcl-2 and PML-RARa, increased the levels of Bax, cleaved caspase-3, cleaved PARP and p-p38MAPK. Meanwhile, SB203580 partially increase the viability of NB4 cells and reduced apoptosis induced by lapatinib; and decreased the levels of p-p38MAPK, Bax, cleaved caspase-3 and cleaved PARP. These results suggested that lapatinib inhibited proliferation and induced apoptosis in NB4 cells, and p38MAPK signal pathway may be involved in these processes.

关 键 词：拉帕替尼 NB4细胞 细胞增殖 细胞凋亡 P38MAPK 

分 类 号：R733.71[医药卫生—肿瘤]