上海交通大学学报(医学版)

• 论著(基础研究) • 上一篇    下一篇

sCD40L对白血病Kasumi-1细胞的影响及机制

任明强,陈琦,章莹   

  1. 遵义医学院附属医院血液科,遵义 563003
  • 出版日期:2017-01-28 发布日期:2017-01-19
  • 作者简介:任明强(1977—),男,副教授,硕士;电子信箱:844198637@qq.com。
  • 基金资助:

    贵州省科技厅联合基金项目[黔科合LH字(2015)7468]

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

REN Ming-qiang, CHEN Qi, ZHANG Ying   

  1. Department of Hematology, Affiliated Hospital of Zunyi Medical College, Zunyi 563003, China
  • Online:2017-01-28 Published:2017-01-19
  • Supported by:

    Jiont Project of Science and Technology Department of Guizhou Province,LH(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4 μg/mL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。

关键词: sCD40L, Kasumi-1细胞, 增殖, 凋亡

Abstract:

Objective · To investigate the effects of soluble CD40 Ligand (sCD40L) on the proliferation, apoptosis of Kasumi-1 cells in vitro and its mechanism. Methods · Different densities of sCD40L were applied to Kasumi-1 cells and the MTT assay was used to observe the inhibiting effects of sCD40L on the cell proliferation at 24, 48 and 72 h to screen the optimum concentration of sCD40L and intervention time. After Kasumi-1 cells were treated with the optimum concentration of sCD40L and time, the flow cytometry (FCM) was adopted to test the apoptosis and cell cycle of Kasumi-1 cells, Western blotting was used to measure the expression of Cytc, Bax, Bid and Bcl-2, ELISA was used to test the expression of IL-17 in the culture supernatants. Results · The optimum sCD40L experimental concentration and intervention time was 4 μg/mL and 48 h. After Kasumi-1 cells were disposed by sCD40L, the cell proliferation was inhibited and the apoptosis was increased significantly. SCD40L could change Kasumi-1 cell cycle distribution which showed increasing the proportion of cells in G1 phase and reducing the proportion of cells S phase. The expression level of proapoptotic factor Bax, Bid, Cytc of Kasumi-1 cell and IL-17 in cell culture supernatant were decreased and anti-apoptotic factor Bcl-2 was increased. Conclusion · SCD40L can significantly suppress Kasumi-1 cells proliferation and induce Kasumi-1 cells apoptosis. The mechanism maybe involve mitochondrial pathways and up-regulation of IL-17 level.

Key words: sCD40L, Kasumi-1 cell, proliferation, apoptosis