DACT3 promotes ferroptosis of glioma cells through activating BMP4/Smad signaling pathway

Li ZHENG; Mei DENG; Liya WANG; Jiqin ZHANG; Jianhe YUE; Guoqiang HAN; Shuo PENG; Jian LIU; Hao YIN; Ying TAN

doi:10.16016/j.2097-0927.202211125

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DACT3 promotes ferroptosis of glioma cells through activating BMP4/Smad signaling pathway

Basic Medicine | 更新时间：2026-01-09

- DACT3 promotes ferroptosis of glioma cells through activating BMP4/Smad signaling pathway
- Journal of Army Medical University Vol. 45, Issue 13, Pages: 1397-1404(2023)
- 作者机构：
  
  1.563000 贵州遵义，遵义医科大学研究生院
  2.550002 贵阳，贵州省人民医院：神经外科
  3.550002 贵阳，贵州省人民医院：麻醉科
- 作者简介：
- 基金信息：
  
  Supported by the Regional Program of National Natural Science Foundation of China (81960454) and the Sci & Tech Foundation of Guizhou Provincial Department of Science and Technology (2020-1Z066)
- DOI：10.16016/j.2097-0927.202211125
  CLC： R394.3;R73-362;R730.264
- Received：17 November 2022，
  
  Revised：2023-01-03，
  
  Published：15 July 2023
- 稿件说明：
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Li ZHENG, Mei DENG, Liya WANG, et al. DACT3 promotes ferroptosis of glioma cells through activating BMP4/Smad signaling pathway[J]. Journal of Army Medical University, 2023, 45(13): 1397-1404.
DOI：

Li ZHENG, Mei DENG, Liya WANG, et al. DACT3 promotes ferroptosis of glioma cells through activating BMP4/Smad signaling pathway[J]. Journal of Army Medical University, 2023, 45(13): 1397-1404. DOI： 10.16016/j.2097-0927.202211125.

摘要

目的

探究基因DACT3促进神经胶质瘤细胞(U251、U118)铁死亡的分子机制。

方法

培养人U251、U118胶质瘤细胞，慢病毒转染构建稳定过表达DACT3的细胞后，qRT-PCR和Western blot检测转染效率。上述细胞分成3组：Ctrl组(对照组)、DACT3组(过表达DACT3细胞组)和DACT3+ferrostatin-1组(过表达DACT3+铁死亡抑制剂)。采用CCK-8检测细胞生存率，铁离子、MDA、谷胱甘肽(glutatione

GSH)检测试剂盒测定细胞铁离子、MDA、GSH含量，Western blot检测GPX4、SLC7A11、TFR1表达。进一步将上述细胞分为以下3组：Ctrl组(对照组)、DACT3组(过表达DACT3细胞组)和DACT3+ siBMP4组(过表达DACT3+敲低BMP4组)，再次利用CCK-8检测细胞生存率，铁离子、MDA、GSH检测试剂盒测定细胞铁离子、MDA、GSH含量，Western blot检测GPX4、SLC7A11、TFR1、BMP4、p-Smad表达。

结果

DACT3组中DACT3 mRNA和蛋白表达均显著高于Ctrl组；与Ctrl组比较，DACT3组细胞生存率明显下降，铁离子、MDA含量显著升高，GSH含量明显下降，GPX4、SLC7A11表达显著下降，TFR1、p-Smad、BMP4表达明显升高

差异均具有统计学意义(

＜0.05)；DACT3+ferrostatin-1组、DACT3+siBMP4组均能一定程度抵消DACT3导致的上述指标的变化，差异具有统计学意义(

＜0.05)。

结论

DACT3可以促进U251、U118胶质瘤细胞铁死亡，其机制与BMP4/SMAD信号通路激活有关。

Abstract

Objective

To investigate the molecular mechanism of DACT3 promoting ferroptosis in human glioma cell lines U251 and U118.

Methods

Glioma U251 and U118 cells were transfected with lentiviral vector carrying DACT3 to establish a cell line with stable over-expression of DACT3

which was verified for the efficiency of transfection by real-time PCR and Western blotting. Then the above cells were divided into control group (Ctrl group)

DACT3 overexpression group (DACT3 group) and DACT3 overexpression+ferroptosis inhibitor group (DACT3+ferrostatin-1 group). CCK-8 assay was applied to detect the cell survival rate

and iron ion

malondialdehyde (MDA) and glutatione (GSH) reagent kits were employed to determine the contents of iron ion

MDA and GSH in the cells. Western blotting was applied to measure the protein levels of GPX4

SLC7A11

TFR1

BMP4

and p-Smad. The above cells were further divided into Ctrl group

DACT3 group and DACT3+siBMP4 group (knockdown of BMP4). Above experiments were performed again for cell survival rate

contents of iron ion

MDA and GSH

and protein levels of above proteins.

Results

The expression of DACT3 at mRNA and protein levels was significantly higher in the DACT3 group than the Ctrl group. The DACT3 group had significantly reduced cell survival rate

increased contents of iron ions and MDA but decreased conten

t of GSH

down-regulation of GPX4 and SLC7A11 and up-regulation of TFR1

p-Smad and BMP4 when compared with the Ctrl group (

0.05). However

ferrostatin-1 treatment and knockdown of BMP4 reversed the changes of the above indicators caused by DACT3 to a certain extent

with statistical significances (

0.05).

Conclusion

DACT3 promotes the ferroptosis of glioma U251 and U118 cells

which may be associated with the activation of BMP4/Smad signaling pathway.

关键词

Keywords

references

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