摘要
目的:探讨佛波酯化合物TPA对急性早幼粒细胞白血病细胞NB4增殖、凋亡的影响及其分子机制。方法:CCK-8法检测不同浓度TPA在不同时间点对NB4细胞增殖的影响;瑞氏-吉姆萨染色观察NB4细胞形态学变化;流式细胞术检测TPA处理后NB4细胞周期、凋亡情况;高通量微阵列分析和实时荧光定量PCR法分析TPA处理后NB4细胞m RNA的表达;Western blot检测CDKN1A、CDKN1B、CCND1、MYC、Bax、Bcl-2、c-Caspase 3、c-Caspase 9、PIK3R6、AKT和p-AKT的蛋白表达。结果:与对照组相比,TPA能抑制NB4细胞增殖,诱导细胞向成熟粒-单核系分化;诱导细胞G_(1)期阻滞及凋亡;差异表达的m RNA显著富集在PI3K/AKT通路;TPA能提高NB4细胞中CCND1、CCNA1、CDKN1A的m RNA水平,降低MYC的m RNA水平;TPA能上调NB4细胞中CDKN1A、CDKN1B、CCND1、Bax、c-Caspase 3、c-Caspase 9、PIK3R6蛋白水平,下调MYC、Bcl-2、p-AKT蛋白水平。结论:TPA通过调控PIK3/AKT信号通路诱导NB4细胞周期阻滞于G_(1)期并促进其凋亡。
Objective:To investigate the effect of phorbol-12-myristate-13-ace-tate(TPA)on the proliferation and apoptosis of acute promyelocytic leukemia cell line NB4 and its molecular mechanism.Methods:The effect of different concentrations of TPA on the proliferation of NB4 cells at different time points was detected by CCK-8 assay.The morphological changes of NB4 cells were observed by Wright-Giemsa staining.The cell cycle and apoptosis of NB4 cells after TPA treatment were detected by flow cytometry.The mRNA expressions of NB4 cells after TPA treatment were analyzed by high-throughput microarray analysis and real-time quantitative PCR.Western blot was used to detect the protein expression of CDKN1A,CDKN1B,CCND1,MYC,Bax,Bcl-2,c-Caspase 3,c-Caspase 9,PIK3R6,AKT and p-AKT.Results:Compared with the control group,TPA could inhibit the proliferation of NB4 cells,induce the cells to become mature granulocyte-monocyte differentiation,and also induce cell G1 phase arrest and apoptosis.Differentially expressed mRNAs were significantly enriched in PI3K/AKT pathway.TPA treatment could increase the mRNA levels of CCND1,CCNA1,and CDKN1A,while decrease the mRNA level of MYC.It could also up-regulate the protein levels of CDKN1A,CDKN1B,CCND1,Bax,c-Caspase 3,c-Caspase 9,and PIK3R6,while down-regulate MYC,Bcl-2,and p-AKT in NB4 cells.Conclusion:TPA induces NB4 cell cycle arrest in G1 phase and promotes its apoptosis by regulating PIK3/AKT signaling pathway.
作者
赵盼
张翀
董雪梅
颜鲁伟
米乐园
李亚娇
康甲超
王晶
ZHAO Pan;ZHANG Chong;DONG Xue-Mei;YAN Lu-Wei;MI Le-Yuan;LI Ya-Jiao;KANG Jia-Chao;WANG Jing(School of Public Health,Gansu University of Chinese Medicine,Lanzhou 730000,Gansu Province,China;Clinical Laboratory Center of Gansu Provincial Maternity and Child Care Hospital,Lanzhou 730050,Gansu Province,China;The First Clinical Medical College of Lanzhou University,Lanzhou 730000,Gansu Province,China)
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2023年第5期1296-1302,共7页
Journal of Experimental Hematology
基金
甘肃省卫生行业科研计划项目(GSWSKY2017-27)
甘肃省高校科研项目一般项目(甘财教【2013】116号)
国家自然科学基金(82060829)。
关键词
TPA
急性早幼粒细胞白血病
NB4细胞
增殖
凋亡
phorbol-12-myristate-13-ace-tate
acute promyelocytic leukemia
NB4 cell
proliferation
apoptosis
