Notch1高表达急性T淋巴细胞白血病发病中细胞增殖的相关机制  

Mechanism of cell proliferation in acute T cell leukemia with high expression of Notch1 gene

作　　者：李香 沈莉菁[1] 陈芳源[1] 高晓 钟华[1] 蔡佳翌[1] 张义炜[1] 朱迪 Li Xiang;Shen Lijing;Chen Fangyuan;Gao Xiao;Zhong Hua;Cai Jiayi;Zhang Yiwei;Zhu Di(Department of Hematology,Renji Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200125,China)

机构地区：[1]上海交通大学医学院附属仁济医院血液科,200125

出　　处：《国际输血及血液学杂志》2018年第5期369-378,共10页International Journal of Blood Transfusion and Hematology

基　　金：国家自然科学基金青年项目（81300405）;上海市科学技术委员会资助项目（12401906700）

摘　　要：目的探讨Notch1高表达急性T淋巴细胞白血病（T-ALL）发病中，细胞增殖的相关机制及靶向药物干预效价。 方法选择Notch1高表达的T-ALL细胞株（A3、MOLT-4、Jurkat细胞株）和来源于20例健康志愿者的外周血单个核细胞（PBMC）为研究对象。采用不同浓度（0、50、100 nmol/L）的细胞周期抑制剂palbociclib处理T-ALL细胞株与PBMC，采用CCK-8法检测其细胞增殖状态，计算不同浓度palbociclib处理时的半数抑制浓度（IC50）值，细胞涂片观察细胞形态，并且采用流式细胞术分析T-ALL细胞株与PBMC的细胞周期变化，实时荧光定量（RT）-PCR检测相关基因mRNA的相对表达水平。采用t检验比较不同浓度palbociclib处理后T-ALL细胞株与PBMC相关基因mRNA的相对表达水平等呈正态分布计量指标。结果①A3、MOLT-4细胞株Notch1基因表达的灰度值百分比，分别为（62.3±3.40）%、（77.6±4.71）%，显著高于PBMC的（20.8±1.53）%，并且差异均有统计学意义（t=6.236、10.034，P=0.025、0.008）；Jurkat细胞Notch1基因表达的灰度值百分比为（28.5±1.36）%，与PBMC比较，差异无统计学意义（t=1.257，P=0.430）。②相较于PBMC，MOLT-4细胞株增殖活性被palbocicib显著抑制，其次为A3细胞株，并且3种细胞增殖活性抑制呈明显时间和浓度依赖。经palbocicib处理48 h后，A3及MOLT-4细胞株的IC50值分别为（0.43±0.12）μmol/L和（0.10±0.06）μmol/L，显著低于PBMC的（0.83±0.15）μmol/L，前二者分别与后者相比，差异均有统计学意义（t=12.354、7.486，P=0.006、0.015）。③细胞形态结果显示，相较于PBMC，经palbocicib 100 nmol/L处理后，A3和MOLT-4细胞株的细胞增殖受到明显抑制，细胞分散，簇状增殖现象消失。④细胞周期检测结果显示，经palbocicib 50 nmol/L处理时S＋G2期A3、MOLT-4细胞株的细胞比例较palbocicib 0 nmol/L处理时显著降低，并且差异均有统计学意义（t=5.358�ObjectiveTo investigate the mechanisms of cell proliferation of acute T cell leukemia （T-ALL） with high expression of Notch1 gene and explore the potential of targeted drug intervention.MethodsA3, MOLT-4, Jurkat cell lines and peripheral blood mononuclear cell （PBMC） derived from 20 healthy volunteers were selected as subjects. All the cells were treated with palbociclib at different concentrations（0, 50, 100 nnol/L）. The cell proliferation status was detected by CCK-8 method. Inhibitory concentration 50 （IC50） values of different concentrations of palbociclib were calculated. The cell morphology was observed, and the cell cycle changes of T-ALL cell line and PBMC were analyzed by flow cytometry. Real-time quantitative （RT）-PCR was used to detect the mRNA relative expression level of related genes. The t-test was used to compare the mRNA relative expression of related genes in T-ALL cells and PBMC after treatment with different concentrations of palbociclib.Results① The gray scale percentages of Notch1 gene expression in A3 and MOLT-4 cell lines were （62.3±3.40）% and （77.6±4.71）%, which were significantly higher than that of PBMC with （20.8±1.53）%. And the differences were significantly （t=6.236, 10.034; P=0.025, 0.008）. There were no significant difference between （28.5±1.36）% of Jurkat cell line and PBMC （t=1.257, P=0. 430）. ② Compared with PBMC, the proliferation activity of MOLT-4 cell line was significantly inhibited by palbociclib, followed by A3 cell line.And that inhibition of cell proliferation activities of all cells was time- and concentration-dependent.After 48 h, the IC50 values of A3 and MOLT-4 cell lines were （0.43±0.12） μmol/L and （0.10±0.16） μmol/L respectively, which were lower than that of PBMC with （0.83±0.15）μmol/L, and the differences were significantly （t=12.354, 7.486; P=0.006, 0.015）. ③ Microscopically, compared with PBMC, the proliferation of A3 and MOLT-4 cell lines were inhibited significantly at 100 nmol/L pal

关 键 词：白血病 T细胞 急性病 细胞增殖 受体 Notch1 Palbociclib 

分 类 号：R733.71[医药卫生—肿瘤]