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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

30卷

期数:

2024年03期

页码:

193-199

栏目:

肿瘤研究专题

出版日期:

2024-05-20

文章信息/Info

Title:

Effects of erastin on the growth and proliferation of osteosarcoma cells

文章编号:

1006-8147(2024)03-0193-07

作者:

郝名英; 刘思琪; 黄欣宇; 耿鑫

（天津医科大学基础医学院生物化学与分子生物学系，天津300070）

Author(s):

HAO Mingying;  LIU Siqi;  HUANG Xinyu;  GENG Xin

（Department of Biochemistry and Molecular Biology， School of Basic Medical Sciences， Tianjin Medical University， Tianjin 300070， China）

关键词:

骨肉瘤; 铁死亡; 端粒替代延长

Keywords:

osteosarcoma; ferroptosis; alternative lengthening of telomeres

分类号:

R34

DOI:

10.20135/j.issn.1006-8147.2024.03.0193

文献标志码:

A

摘要:

目的：探究erastin对骨肉瘤（OS）细胞生长、增殖的影响及其分子机制。方法：使用STRING数据库构建OS易突变蛋白、铁死亡相关蛋白和端粒相关蛋白的相互作用网络。检测 2.5 μmol/L erastin处理OS细胞后铁死亡相关指标的变化、端粒替代延长（ALT）相关表型的变化、端粒损伤情况、端粒保护蛋白的变化和对OS细胞生长增殖、迁移的影响。结果：STRING蛋白相互作用网络分析显示，铁死亡相关蛋白、OS中突变蛋白和端粒相关蛋白相互作用网络包括RPA1、BARD1、UBE3A、AURKA、MDM2、SIRT1、CREBBP、IGFBP5、DDX5及EP300。2.5 μmol/L erastin处理OS细胞后，Western印迹显示谷胱甘肽过氧化物酶（GPX）4、SLC家族氨基酸转运蛋白（SLC7A11）表达量明显降低（t=13.24、4.399，均P<0.05），毛细血管扩张性共济失调和Rad3相关蛋白（ATR）表达量明显降低（t=10.68，P<0.001），端粒重复结合因子（TRF）1、2表达量明显升高（t=8.006、10.79，均P<0.01）； 丙二醛（MDA）检测显示脂质过氧化物含量升高（t=2.951，P<0.05）；活性氧簇（ROS）检测试剂盒显示，ROS水平明显增加（q=11.03，P<0.001）；ALT相关表型结果显示，c-circle水平明显降低（t=16.20，P<0.000 1），早幼粒细胞白血病核小体（PML）与端粒的定位明显降低（t=6.018，P<0.01）；免疫荧光结果显示，53BP1、γ-H2AX与端粒的定位明显增加（q=4.391、4.653，均P<0.05）；CCK8、克隆形成实验结果显示，细胞生长增殖水平降低（t=10.86、4.972，均P<0.01）；划痕实验结果表明，细胞的迁移水平降低（t=2.953，P<0.05）。结论：伴随铁死亡的发生，erastin可以减弱ALT并抑制OS细胞的生长、增殖和迁移。

Abstract:

Objective： To explore the effect of erastin on the growth and proliferation of osteosarcoma （OS） cells and its molecular mechanism. Methods： The interaction network of mutant protein with ferroptosis associated protein and telomere related protein in OS was constructed using STRING database. The effects of 2.5 μmol/L erastin treatment on ferroptosis associated protein， alternative lengthening of telomeres （ALT） related phenotype， telomere damage， telomere protective proteins， and growth， proliferation and migration of OS cells were detected. Results： The STRING protein interaction network analysis showed that ferroptosis related proteins， mutant proteins in OS， and telomere related protein interaction networks included RPA1， BARD1， UBE3A， AURKA， MDM2， SIRT1， CREBBP， IGFBP5， DDX5 and EP300. After 2.5 μmol/L erastin treatment， Western blotting showed that glutathione peroxidase 4 （GPX4） and SLC family amino acid transporter （SLC7A11） protein expression decreased significantly （t=13.24，4.399， both P<0.05）. Ataxia telangiectasia and Rad3-related protein （ATR） expression was significantly decreased （t=10.68， P<0.001）. The expression levels of shelterin telomere repeat binding 1 and 2 （TRF1 and TRF2） proteins were significantly increased （t=8.006， 10.79， both P<0.01）. Malondialdehyde （MDA） detection results showed that lipid peroxide content increased （t=2.951， P<0.05）， and reactive oxygen species （ROS） detection kit results showed that ROS level increased significantly （q=11.03， P<0.001）. The results of ALT related phenotype showed that c-circle level was significantly decreased （t=16.20， P<0.000 1）， promyelocytic leukemia （PML） and telomere localization were significantly decreased （t=6.018， P<0.01）. Immunofluorescence results showed that the localization of 53BP1， γ-H2AX and telomeres increased significantly （q=4.391， 4.653， both P<0.05）. CCK8 and the results of clonal formation experiment showed that the cell growth and proliferation level decreased （t=10.86， 4.972，both P<0.01）， the scratch test results showed that the migration level of the cells was reduced（t=2.953，P<0.05）. Conclusion：With the occurrence of ferroptosis，erastin can reduce ALT and inhibit the growth，proliferation and migration of OS cells.

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备注/Memo

备注/Memo:

基金项目 天津市教委科研计划（2021ZD037）
作者简介 郝名英（1997-），男，硕士在读，研究方向：医学生物化学与分子生物学；通信作者：耿鑫，E-mail：gengx@tmu.edu.cn。

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更新日期/Last Update: 2024-05-20