sCD40L对白血病Kasumi-1细胞的影响及机制  

Effects of soluble CD40 ligand on leukemia Kasumi-1 cells and its mechanism

作　　者：任明强[1] 陈琦[1] 章莹[1] REN Ming-qiang;CHEN Qi;ZHANG Ying(Department of Hematology, Affiliated Hospital of Zunyi Medical College, Zunyi 563003, China)

机构地区：[1]遵义医学院附属医院血液科,遵义563003

出　　处：《上海交通大学学报（医学版）》2017年第1期21-25,共5页Journal of Shanghai Jiao tong University:Medical Science

基　　金：贵州省科技厅联合基金项目[黔科合LH字(2015)7468]~~

摘　　要：目的·探讨可溶性CD40配体(sCD40L)对体外培养白血病Kasumi-1细胞增殖、凋亡的影响及分子机制。方法·用不同梯度浓度sCD40L处理Kasumi-1细胞,在24、48、72 h测定细胞增殖率,以筛选优化的干预浓度和时间。用优化的sCD40L浓度和时间干预处理Kasumi-1细胞后,采用流式细胞术检测细胞凋亡率和细胞周期分布;采用免疫印迹法(Western blotting)检测凋亡因子Cytc、Bax、Bid和Bcl-2的表达;采用ELISA法检测sCD40L处理后Kasumi-1细胞培养上清液中IL-17含量。结果·sCD40L体外最佳干预实验浓度和时间分别为4μg/m L和48 h。经sCD40L处理后,Kasumi-1细胞体外增殖明显受到抑制,同时凋亡率明显升高;细胞周期分布表现为G1期细胞比例增加、S期细胞比例降低;促凋亡因子Bax、Bid、Cytc表达水平明显升高,抗凋亡因子Bcl-2表达水平显著降低;Kasumi-1细胞培养上清液中IL-17含量明显增高。结论·sCD40L可显著抑制Kasumi-1细胞增殖并促进其凋亡,其作用机制与线粒体通路及上调IL-17表达水平有关。Objective·To investigate the effects of soluble CD40Ligand(sCD40L)on the proliferation,apoptosis of Kasumi-1cells in vitro and itsmechanism.Methods·Different densities of sCD40L were applied to Kasumi-1cells and the MTT assay was used to observe the inhibiting effects ofsCD40L on the cell proliferation at24,48and72h to screen the optimum concentration of sCD40L and intervention time.After Kasumi-1cells weretreated with the optimum concentration of sCD40L and time,the flow cytometry(FCM)was adopted to test the apoptosis and cell cycle of Kasumi-1cells,Western blotting was used to measure the expression of Cytc,Bax,Bid and Bcl-2,ELISA was used to test the expression of IL-17in the culturesupernatants.Results·The optimum sCD40L experimental concentration and intervention time was4μg/mL and48h.After Kasumi-1cells weredisposed by sCD40L,the cell proliferation was inhibited and the apoptosis was increased significantly.SCD40L could change Kasumi-1cell cycledistribution which showed increasing the proportion of cells in G1phase and reducing the proportion of cells S phase.The expression level of proapoptoticfactor Bax,Bid,Cytc of Kasumi-1cell and IL-17in cell culture supernatant were decreased and anti-apoptotic factor Bcl-2was increased.Conclusion·SCD40L can significantly suppress Kasumi-1cells proliferation and induce Kasumi-1cells apoptosis.The mechanism maybe involvemitochondrial pathways and up-regulation of IL-17level.

关 键 词：SCD40L KASUMI-1细胞 增殖 凋亡 

分 类 号：R733.7[医药卫生—肿瘤]