Depletion of F-actin binding domain reduces the ability of BCR/ABL to resist apoptosis in 293T cells

Fangzhu ZHOU; Zhenglan HUANG; Zhenhong LUO; Teng WANG; Wenli FENG

doi:10.16016/j.1000-5404.201802024

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Depletion of F-actin binding domain reduces the ability of BCR/ABL to resist apoptosis in 293T cells

更新时间：2026-01-09

- Depletion of F-actin binding domain reduces the ability of BCR/ABL to resist apoptosis in 293T cells
- Journal of Third Military Medical University Vol. 40, Issue 17, Pages: 1548-1553(2018)
- 作者机构：
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81572060);the Postgraduate Innovation Project of Chongqing(CYS16136)
- DOI：10.16016/j.1000-5404.201802024
  CLC：
- Received：04 February 2018，
  
  Published：15 September 2018
- 稿件说明：
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Fangzhu ZHOU, Zhenglan HUANG, Zhenhong LUO, et al. Depletion of F-actin binding domain reduces the ability of BCR/ABL to resist apoptosis in 293T cells[J]. Journal of Third Military Medical University, 2018, 40(17): 1548-1553.
DOI：

Fangzhu ZHOU, Zhenglan HUANG, Zhenhong LUO, et al. Depletion of F-actin binding domain reduces the ability of BCR/ABL to resist apoptosis in 293T cells[J]. Journal of Third Military Medical University, 2018, 40(17): 1548-1553. DOI： 10.16016/j.1000-5404.201802024.

摘要

Abstract

Objective

To investigate the effects of F-actin binding domain (FABD) on BCR/ABL function in 293T cells by transfecting wild-type pAd-Bcr/Abl and mutant pAd-Bcr/Abl -ΔFABD.

Methods

Firstly

293T cells were transfected wild-type pAd-Bcr/Abl and mutant pAd-Bcr/Abl-ΔFABD plasmid

respectively. Then the transfected cells and blank control cells were cultivated for 24

or 72 h. Flow cytometry was used to detect the fluorescence intensity to screen optimal transfection conditions. Western blotting was employed to detect the expression of related proteins. Secondly

the effects of FABD on proliferation and apoptosis of the cells promoted by BCR/ABL oncoprotein were detected by WST-1 assay and flow cytometry

respectively. DAPI staining was adopted to observe the changes of nuclear morphology. The protein expression of p-BCR/ABL

p-AKT

p-ERK

and apoptosis related protein Caspase-3

and PARP activity was determined by Western blotting.

Results

The results of flow cytometry showed that the efficiency of plas

mid transfection was increased in a time-dependent manner

and 48 h was identified as the optimal transfection time. Western blotting results confirmed the pAd-Bcr/Abl and mutant pAd-Bcr/Abl-ΔFABD plasmids were constructed successfully. After the transfection of pAd-Bcr/Abl and mutant pAd-Bcr/Abl-ΔFABD plasmids into 293T cells

WST-1 assay and flow cytometry that FABD depletion significant inhibited cell proliferation and promoted apoptosis

with the percentage of apoptotic cells accounting for 29.0% in 48 h after treatment (

0.05). The depletion also down-regulated the expression of p-AKT and p-ERK and activated Caspase-3 and PARP in 48 h after transfection.

Conclusion

FABD depletion reduces the cell apoptosis promoted by BCR/ABL oncoprotein

which might be though down-regulating AKT and ERK signaling pathways and activating mitochondrial associated apoptotic pathway.

关键词

Keywords

references

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