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Mdm2过表达诱导ALK阳性非小细胞肺癌克唑替尼耐药的实验研究
Overexpression of Mdm2 induces crizotinib resistance in targeted therapy for ALK-positive NSCLC
- 陆军军医大学学报 2022年44卷第24期 页码：2500-2506
- 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：
[叶明翔，E-mail：mingxiangye@gmail.com YE Mingxiang, E-mail: mingxiangye@gmail.com
- 基金信息：中国博士后科学基金项目(2020M670093ZX)
DOI：10.16016/j.2097-0927.202209006
中图分类号： R394.6;R734.2;R915收稿：2022-09-01，
修回：2022-10-24，
纸质出版：2022-12-30
- 稿件说明：
移动端阅览
刘嘉欣, 李姗姗, 王栋, 等. Mdm2过表达诱导ALK阳性非小细胞肺癌克唑替尼耐药的实验研究[J]. 陆军军医大学学报, 2022,44(24):2500-2506.
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.
刘嘉欣, 李姗姗, 王栋, 等. Mdm2过表达诱导ALK阳性非小细胞肺癌克唑替尼耐药的实验研究[J]. 陆军军医大学学报, 2022,44(24):2500-2506. DOI： 10.16016/j.2097-0927.202209006.
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
SODA M, CHOIY L, ENOMOTO M, et al . Identification of the transforming EML 4- ALK fusion gene in non-small-cell lung cancer[J ] . Nature, 2007, 448(7153): 561-566. DOI: 10.1038/nature05945..
TAKEUCHI K, CHOIY L, TOGASHI Y, et al . KIF 5 B-ALK, a novel fusion oncokinase identified by an immunohistochemistry-based diagnostic system for ALK-positive lung cancer[J ] . Clin Cancer Res, 2009, 15(9): 3143-3149. DOI: 10.1158/1078-0432.CCR-08-3248..
SASAKI T, RODIGS J, CHIRIEAC L R, et al . Thebiology and treatment of EML4-ALK non-small cell lung cancer[J ] . Eur J Cancer, 2010, 46(10): 1773-1780. DOI: 10.1016/j.ejca.2010.04.002.
SOLOMONB J, MOK T, KIM D W, et al . First-line crizotinib versus chemotherapy in ALK-positive lung cancer[J ] . N Engl J Med, 2014, 371(23): 2167-2177. DOI: 10.1056/NEJMoa1408440.
CHOIY L, SODA M, YAMASHITA Y, et al . EML4-ALK mutations in lung cancer that confer resistance to ALK inhibitors[J ] . N Engl J Med, 2010, 363(18): 1734-1739. DOI: 10.1056/NEJMoa1007478.
SALIDO M, PIJUAN L, MARTÍNEZ-AVILÉS L, et al . Increased ALK gene copy number and amplification are frequent in non-small cell lung cancer[J ] . J Thorac Oncol, 2011, 6(1): 21-27. DOI: 10.1097/JTO.0b013e3181fb7cd6..
SHANGARY S, WANG S M. Targeting the MDM2-p53 interaction for cancer therapy[J]. Clin Cancer Res, 2008, 14 (17): 5318-5324. DOI: 10.1158/1078-0432.CCR-07-5136.
CHÈNE P. Inhibiting the p53-MDM2 interaction: an important target for cancer therapy[J]. Nat Rev Cancer, 2003, 3(2): 102-109. DOI: 10.1038/nrc991.
MIYACHI M, KAKAZU N, YAGYU S, et al . Restoration of p53 pathway by nutlin-3 induces cell cycle arrest and apoptosis in human rhabdomyosarcoma cells[J ] . Clin Cancer Res, 2009, 15(12): 4077-4084. DOI: 10.1158/1078-0432.CCR-08-2955.
PETERS S, CAMIDGE D R, SHAW A T, et al . Alectinib versus crizotinib in untreated ALK-positive non-small-cell lung cancer[J ] . N Engl J Med, 2017, 377(9): 829-838. DOI: 10.1056/NEJMoa1704795.
SHAWA T, BAUER T M, DE MARINIS F, et al . First-line lorlatinib or crizotinib in advanced ALK-positive lung cancer[J ] . N Engl J Med, 2020, 383(21): 2018-2029. DOI: 10.1056/NEJMoa2027187.
DEBS P. Cell cycle regulatory functions of the human oncoprotein MDM2[J]. Mol Cancer Res, 2003, 1(14): 1009-1016.
DWORAKOWSKA D, JASSEM E, JASSEM J, et al . MDM 2 gene amplification: a new independent factor of adverse prognosis in non-small cell lung cancer (NSCLC)[J ] . Lung Cancer, 2004, 43(3): 285-295. DOI: 10.1016/j.lungcan.2003.09.010..
SENTURKJ C, BOHLMAN S, MANFREDI J J. Mdm2 selectively suppresses DNA damage arising from inhibition of topoisomerase Ⅱ independent of p53[J]. Oncogene, 2017, 36(44): 6085-6096. DOI: 10.1038/onc.2017.229.
LAUL M S, NUGENT J K, ZHAO X, et al . HDM2 antagonist Nutlin-3 disrupts p73-HDM2 binding and enhances p73 function[J ] . Oncogene, 2008, 27(7): 997-1003. DOI: 10.1038/sj.onc.1210707.
VAN MAERKEN T, FERDINANDE L, TAILDEMAN J, et al . Antitumor activity of the selective MDM2 antagonist nutlin-3 against chemoresistant neuroblastoma with wild-type p53[J ] . J Natl Cancer Inst, 2009, 101(22): 1562-1574. DOI: 10.1093/jnci/djp355.
DONGRE A, WEINBERGR A. New insights into the mechanisms of epithelial-mesenchymal transition and implications for cancer[J]. Nat Rev Mol Cell Biol, 2019, 20(2): 69-84. DOI: 10.1038/s41580-018-0080-4.
YANG J, WEINBERGR A. Epithelial-mesenchymal transition: at the crossroads of development and tumor metastasis[J]. Dev Cell, 2008, 14(6): 818-829. DOI: 10.1016/j.devcel.2008.05.009.
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