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Journal of Cancer Research and Clinical Oncology

, Volume 144, Issue 8, pp 1413–1422 | Cite as

Loss of EGFR confers acquired resistance to AZD9291 in an EGFR-mutant non-small cell lung cancer cell line with an epithelial–mesenchymal transition phenotype

  • Jing Xu
  • Xiaoting Zhao
  • Dengfeng He
  • Jinghui Wang
  • Weiying Li
  • Yinghui Liu
  • Li Ma
  • Mei Jiang
  • Yu Teng
  • Ziyu Wang
  • Meng Gu
  • Jianbin Wu
  • Yue Wang
  • Wentao Yue
  • Shucai Zhang
Original Article – Cancer Research
  • 230 Downloads

Abstract

Purpose

AZD9291 is an irreversible, small-molecule inhibitor which has potency against mutant EGFR- and T790M-resistant mutation. Despite the encouraging efficacy in clinical, the acquired resistance will finally occur. Further study will need to be done to identify the acquired resistance mechanisms and determine the next treatment.

Methods

We established an AZD9291-resistant cell line (HCC827/AZDR) from parental HCC827 cell line through stepwise pulsed selection of AZD9291. The expression of EGFR and its downstream pathways were determined by western blot analysis or immunofluorescence assay. The sensitivity to indicated agents were evaluated by MTS.

Results

Compared with parental HCC827 cells, the HCC827/AZDR cells showed high resistance to AZD9291 and other EGFR-TKIs, and exhibited a mesenchymal-like phenotype. Almost complete loss of EGFR expression was observed in HCC827/AZDR cells. But the activation of downstream pathway, MAPK signaling, was found in HCC827/AZDR cells even in the presence of AZD9291. Inhibition of MAPK signaling had no effect on cell viability of HCC827/AZDR and could not reverse AZD9291 resistance because of the subsequent activation of AKT signaling. When treated with the combination of AKT and MAPK inhibitor, HCC827/AZDR showed remarkable growth inhibition.

Conclusions

Loss of EGFR could be proposed as a potential acquired resistance mechanism of AZD9291 in EGFR-mutant NSCLC cells with an EMT phenotype. Despite the loss of EGFR, the activation of MAPK pathway which had crosstalk with AKT pathway could maintain the proliferation and survival of resistant cells. Blocking MAPK and AKT signaling may be a potential therapeutic strategy following AZD9291 resistance.

Keywords

AZD9291 Resistance EGFR loss NSCLC Potential therapy 

Abbreviations

BSA

Bovine serum albumin

BTD

Breakthrough therapy designation

CI

Combination index

ECL

Enhanced chemiluminescence

EGFR

Epidermal growth factor receptor

EMT

Epithelial–mesenchymal transition

FBS

Fetal bovine serum

MAPK

Mitogen-activated protein kinases

NSCLC

Non-small cell lung cancer

ORR

Overall response rate

PFS

Progression-free survival

AKT(PKB)

Protein Kinase B

QOL

Quality of life

TKIs

Tyrosine kinase inhibitors

Notes

Acknowledgements

We would like to thank Beta Pharma (Zhejiang, China) and Shihe Gene Bio-Technology (Nanjing, China) for some technical assistance.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest. The authors are responsible for the content and writing of the paper.

Research involving human participants and/or animals

This article does not contain studies of human participants or animals performed by any of the authors.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Medical Oncology, Beijing Tuberculosis and Thoracic Tumor Research Institute/Beijing Chest HospitalCapital Medical UniversityBeijingChina
  2. 2.Department of Cellular and Molecular Biology, Beijing Tuberculosis and Thoracic Tumor Research Institute/Beijing Chest HospitalCapital Medical UniversityBeijingChina
  3. 3.Department of Traditional Chinese medicine263 Clinical Department of General Hospital of Beijing Military RegionBeijingChina
  4. 4.Central Laboratory, Beijing Obstetrics and Gynecology HospitalCapital Medical UniversityBeijingChina

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