Overexpression of Mdm2 induces crizotinib resistance in targeted therapy for ALK-positive NSCLC

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Overexpression of Mdm2 induces crizotinib resistance in targeted therapy for ALK-positive NSCLC

Monographic Reports: Targeted therapy of lung cancer | 更新时间：2026-01-09

- Overexpression of Mdm2 induces crizotinib resistance in targeted therapy for ALK-positive NSCLC
- Journal of Army Medical University Vol. 44, Issue 24, Pages: 2500-2506(2022)
- 作者机构：
  
  210002 南京，东部战区总医院呼吸与危重症医学科
- 作者简介：
- 基金信息：
  
  the Post-Doctoral Scientific Research Foundation of China(2020M670093ZX)
- DOI：10.16016/j.2097-0927.202209006
  CLC： R394.6;R734.2;R915
- Received：01 September 2022，
  
  Revised：2022-10-24，
  
  Published：30 December 2022
- 稿件说明：
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Jiaxin LIU, Shanshan LI, Dong WANG, et al. Overexpression of Mdm2 induces crizotinib resistance in targeted therapy for ALK-positive NSCLC[J]. Journal of Army Medical University, 2022, 44(24): 2500-2506.
DOI：

Jiaxin LIU, Shanshan LI, Dong WANG, et al. Overexpression of Mdm2 induces crizotinib resistance in targeted therapy for ALK-positive NSCLC[J]. Journal of Army Medical University, 2022, 44(24): 2500-2506. DOI： 10.16016/j.2097-0927.202209006.

摘要

目的

2

探究Mdm2过表达在间变性淋巴瘤激酶(anaplastic lymphoma kinase

ALK)突变的非小细胞肺癌(non-small cell lung cancer，NSCLC)H3122细胞中对克唑替尼敏感性的影响。

方法

2

通过转染Mdm2质粒构建过表达Mdm2蛋白的H3122 Mdm2过表达组细胞，对照组转染空载体构建H3122空载组细胞，给予不同浓度克唑替尼处理后进行MTT、平板克隆、TUNEL染色等实验评估Mdm2表达状态对ALK靶向药物克唑替尼敏感性的影响。通过Western blot实验探究过表达Mdm2后对ALK及其下游信号通路转导以及上皮间质转化过程(epithelial mesenchymal transition

EMT) 标记物的影响。

结果

2

与H3122空载组细胞相比，H3122 Mdm2过表达组细胞出现克唑替尼耐药表型，对药物诱导的细胞凋亡耐受，且过表达Mdm2并不能维持ALK及其下游分子的磷酸化水平。过表达Mdm2后，细胞发生EMT，获得间质细胞特征。

结论

2

过表达Mdm2可能通过诱导EMT过程导致H3122细胞对克唑替尼耐药。

Abstract

Objective

2

To investigate the effect of overexpression of murine double minute 2 (Mdm2) on the sensitivity to crizotinib in the anaplastic lymphoma kinase (ALK)-positive non-small cell lung cancer (NSCLC) cell line H3122.

Methods

2

H3122 cells with Mdm2 overexpression were established by transfection of plasmid encoding Mdm2

and the H3122 cells transfected with empty vector (EV) served as control cells. After the Mdm2 overexpression cells and control cells were treated with different concentrations of crizotinib

an ALK inhibitor

MTT assay

colony formation assay and TUNEL assay were applied to evaluate the sensitivity to crizotinib. Western blotting was used to detect the expression of ALK

its downstream proteins and biomarkers in epithelial mesenchymal transition (EMT).

Results

2

Compared with the control cells

the H3122 cells with Mdm2 overexpression were resistant to crizotinib treatment

and were tolerant to crizotinib-induced apoptosis. Biochemical analysis of signaling transduction suggested that overexpression of Mdm2 failed to restore the phosphorylation of ALK and downstream molecules. Overexpression of Mdm2 also resulted in EMT and acquisition of mesenchymal phenotype in H3122 cells.

Conclusion

2

Overexpression of Mdm2 leads to resistance to crizotinib in H3122 cells probably through the induction of EMT process.

关键词

Keywords

references

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