地西他滨诱导人急性淋巴细胞白血病Jurkat细胞凋亡机制的研究  被引量:2

Study on the mechanism of apoptosis induced by decitabine in human acute lymphoblastic leukemia Jurkat cells

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作  者:张刚 高晓慧 吴海兵 赵晓燕 颜敏超 李园 曾惠 郭晓珺 ZHANG Gang;GAO Xiaohui;WU Haibing;ZHAO Xiaoyan;YAN Minchao;LI Yuan;ZENG Hui;GUO Xiaojun(Department of Hematology,the First Affiliated Hospital of Jiaxing University,Jiaxing 314000,China;Department of Pediatrics,the First Affiliated Hospital of Jiaxing University,Jiaxing 314000,China)

机构地区:[1]嘉兴学院第一附属医院血液科,浙江嘉兴314000 [2]嘉兴学院第一附属医院儿科,浙江嘉兴314000

出  处:《中国现代医生》2020年第1期26-30,I0002,共6页China Modern Doctor

基  金:浙江省医药卫生科技计划项目(2019KY693)

摘  要:目的 研究地西他滨诱导人急性淋巴细胞白血病Jurkat细胞凋亡的机制。方法 常规体外培养人急性淋巴细胞白血病Jurkat细胞株,取对数生长期的细胞传代24 h后加入不同浓度(0.5、1.0、5.0及10.0μmol/L)地西他滨处理,分别继续培养24、48、72 h后,用CCK-8法测定细胞增殖活性;倒置显微镜下观察细胞形态;Annexin V-FITC/PI染色流式细胞术检测地西他滨作用后人急性淋巴细胞白血病Jurkat细胞株的凋亡率;应用透射电镜观察人急性淋巴细胞白血病Jurkat细胞发生凋亡的形态学特征;实时荧光定量Real time-PCR法检测凋亡相关基因Caspase-3、Bcl-2在mRNA水平表达变化的情况。结果 地西他滨可以抑制Jurkat细胞增殖,并且呈浓度和时间依赖性(P<0.05)。倒置显微镜观察到地西他滨作用后细胞形态发生了显著改变;流式细胞术检测细胞凋亡的结果表明地西他滨能够诱导人急性淋巴细胞白血病Jurkat细胞凋亡并且呈浓度依赖性(P<0.05);透射电镜观察结果证明地西他滨可以诱导人急性淋巴细胞白血病Jurkat细胞出现凋亡性细胞死亡,5.0μmol/L地西他滨作用Jurkat细胞72 h后出现大量凋亡小体。另外,Real time-PCR实验研究结果发现地西他滨能够上调促凋亡基因Caspase-3的表达,下调凋亡抑制基因Bcl-2的表达。结论 地西他滨能通过上调促凋亡基因Caspase-3的表达和下调凋亡抑制基因Bcl-2的表达诱导细胞凋亡。Objective To study the mechanism of apoptosis induced by decitabine in human acute lymphoblastic leukemia Jurkat cells.Methods Human acute lymphoblastic leukemia Jurkat cell line was routinely cultured in vitro.cells were harvested in logarithmic growth phase,and then treated with different concentrations(0.5,1.0,5.0 and 10.0μmol/L)of decitabine after passage for 24 h.The cells were cultured continuously for 24,48,and 72 hours respectively.The cell proliferation activity was measured by CCK-8 method.Cell morphology was observed under inverted microscope.Annexin V-FITC/PI staining flow cytometry was used to detect the apoptosis rate of human acute lymphoblastic leukemia Jurkat cell line after decitabine treatment.The morphological characteristics of apoptosis in human acute lymphoblastic leukemia Jurkat cells were observed by transmission electron microscopy.Real-time fluorescence quantitative PCR was used to detect the expression of apoptosis-related genes Caspase-3 and Bcl-2.Results Decitabine inhibited Jurkat cell proliferation in a concentration-and time-dependent manner(P<0.05).Inverted microscopy showed significant changes in cell morphology after decitabine treatment.Flow cytometry detection of apoptosis showed that decitabine can induce apoptosis in human acute lymphoblastic leukemia Jurkat cells in a concentration-dependent manner(P<0.05).Transmission electron microscopy showed that decitabine can induce apoptotic cell death in human acute lymphoblastic leukemia Jurkat cells,and a large number of apoptotic bodies appeared in Jurkat cells treated with 5.0μmol/L decitabine for 72 h.In addition,real time-PCR experimental results showed that decitabine can up-regulate the expression of the proapoptotic gene Caspase-3 and down-regulate the expression of the apoptosis-inhibiting gene Bcl-2.Conclusion Decitabine can induce apoptosis of cells by up-regulating the expression of apoptosis-promoting gene Caspase-3 and down-regulating the expression of apoptosis-inhibiting gene Bcl-2.

关 键 词:地西他滨 急性淋巴细胞白血病 JURKAT细胞 凋亡 

分 类 号:R733.7[医药卫生—肿瘤]

 

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