Induction of apoptosis in TF-1 cell line by bortezomib and its influence on the expression of SALL4 gene

WANG Lan; YU Yi-chuan; FU Lei-hua; HU Cheng-ling; CHEN Lin

doi:10.13406/j.cnki.cyxb.2011.07.017

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Induction of apoptosis in TF-1 cell line by bortezomib and its influence on the expression of SALL4 gene

更新时间：2024-01-17

- Induction of apoptosis in TF-1 cell line by bortezomib and its influence on the expression of SALL4 gene
- Journal of Chongqing Medical University Vol. 36, Issue 7, Pages: 785-789(2011)
- 作者机构：
  
  重庆医科大学附属第二医院血液科
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2011.07.017
  CLC： R733.7
- Published：2011，
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[1]王兰,喻依川,傅雷华,胡成琳,陈林.硼替佐米诱导TF-1细胞凋亡及其对SALL4基因表达的影响[J].重庆医科大学学报,2011,36(07):785-789.

WANG Lan, YU Yi-chuan, FU Lei-hua, et al. Induction of apoptosis in TF-1 cell line by bortezomib and its influence on the expression of SALL4 gene. [J]. Journal of Chongqing Medical University 36(7):785-789(2011)
[1]王兰,喻依川,傅雷华,胡成琳,陈林.硼替佐米诱导TF-1细胞凋亡及其对SALL4基因表达的影响[J].重庆医科大学学报,2011,36(07):785-789. DOI： 10.13406/j.cnki.cyxb.2011.07.017.

WANG Lan, YU Yi-chuan, FU Lei-hua, et al. Induction of apoptosis in TF-1 cell line by bortezomib and its influence on the expression of SALL4 gene. [J]. Journal of Chongqing Medical University 36(7):785-789(2011) DOI： 10.13406/j.cnki.cyxb.2011.07.017.

摘要

目的:探讨蛋白酶体抑制剂硼替佐米（Bortezomib）诱导人红系白血病细胞株TF-1凋亡的作用

及其对Sall4基因表达水平的影响。方法:（1）MTT法检测细胞增殖抑制率。（2）光学显微镜观察细胞的形态学改变。（3）电镜观察超微结构变化。（4）流式细胞仪检测Annexin V-FITC细胞凋亡率。（5）流式细胞仪分析细胞周期。（6）RT-PCR检测Sall4 mRNA的表达水平。结果:（1）硼替佐米呈时间和剂量依赖性抑制TF-1细胞生长

与对照组相比

差异有统计学意义（P<0.05）。24 h和48 h的药物半数抑制浓度（Halfmaximal inhibitory concentration of a substance

IC50）分别为52 nmol/L和27 nmol/L。（2）显微镜下观察细胞皱缩变小

细胞密度明显下降。（3）电镜下观察可见凋亡小体。（4）Annexin V-FITC检测显示随着硼替佐米作用浓度的增加

凋亡率增加

呈剂量依赖关系（P<0.05）。（5）流式细胞仪分析细胞周期

随着硼替佐米作用时间的增加

G2/M期细胞比例逐渐增高

而G0/G1和S期细胞比例则逐渐减少

呈时间依赖关系（P<0.05）。（6）RT-PCR结果示Sall4基因在TF-1细胞上高表达

并随着硼替佐米作用浓度和时间的增加

Sall4 mRNA与β-actin mRNA表达量的积分光密度比值（Integral optical density ratio

V/B）水平下降（P<0.05）。结论:硼替佐米能抑制TF-1细胞增殖

诱导其凋亡

呈时间-剂量依赖关系。Sall4基因在TF-1细胞株上高表达

并随着硼替佐米作用浓度和时间的增加表达水平下降

呈时间-剂量依赖关系。

Abstract

Objective:To investigate the effect of bortezomib on proliferation and apoptosis of human erythroleukemia cells TF-1

and its influence on the expression of Sall4 mRNA.Methods:(1)The level of cell proliferation was analyzed by MTT assay.(2)Morphological changes were observed by microscope.(3)Morphological changes were observed by electron microscope.(4)Flow cytometry with Annexin-V staining was used to analyze cell apoptosis rate.(5)Flow cytometry was used to analyze and cell cycle distribution.(6)The level of the expression of Sall4 mRNA was detected by RT-PCR.Results:(1)Bortezomib inhibited TF-1 cells in a time-and-does dependent manner.The IC50 at 24 and 48 h were 52 nmol/L and 27 nmol/L respectively.(2)Under microscope

it could be observed that the density of cells declined while cell fragments increased.(3)Moreover

under electron microscope

there appeared some apoptotic bodies.(4)Bortezomib could induce apoptosis of TF-1 cells in a dose-dependent manner.(5)At the same time

it caused an increase in G2/M phase and a marked decrease in G0/G1 and S phases in a time-dependent manner.(6)It was found by RT-PCR that the level of Sall4 mRNA was down-regulated with the treatment of bortemomib.Conclusion:Bortezomig can cause apoptosis of TF-1 cells and down-regulate the expression of Sall4 mRNA in a time-and-does dependent manner.

关键词

硼替佐米TF-1细胞株细胞凋亡Sall4基因

Keywords

bortezomibTF-1 cell linecell apoptosisSall4 gene

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