Mechanism of MALAT1 induced osimertinib resistance in HCC827 lung cancer cells
10.3760／cma.j.issn.0253?3766.2019.04.004
- VernacularTitle:肺腺癌转移相关转录本1诱导肺癌HCC827细胞对奥希替尼耐药的作用机制
- Author:
Xiaohong KANG
1
;
Yuanyuan GAO
;
Ying WANG
;
Yanhui CUI
;
Kelei ZHAO
;
Weizheng KOU
;
Zhanhui MIAO
;
Fei CAO
;
Yabin GONG
Author Information
1. 河南省新乡医学院第一附属医院肿瘤科453100
- Keywords:
[Subject words] Lung neoplasms;
Metastasis associated in lung adenocarcinoma transcript;
Epidermal growth factor receptor;
Osimertinib
- From:
Chinese Journal of Oncology
2019;41(4):257-262
- CountryChina
- Language:Chinese
-
Abstract:
Objective To test the effect of metastasis associated in lung adenocarcinoma transcript 1 ( MALAT1) and／or osimertinib on the proliferation and apoptosis of HCC827 cells, and explore the potential mechanism of MALAT1 induced resistance to osimertinib. Methods We transfected HCC827 cells with LV?vector or LV?over／MALAT1. Stable transfected cells ( HCC827／Vector, HCC827／MALAT1) were selected by adding puromycin. HCC827／MALAT1 cells were further transfected with the shRNA?negative control ( NC) or shRNA?human epidermal growth factor receptor 3 ( ERBB3 ) plasmid. The effects of overexpression of MALAT1, knockdown of ERBB3 and／or osimertinib on the proliferation of HCC827 cells were evaluated by 3?(4,5?dimethyl?2?thiazolyl)?2,5?diphenyl?2H tetrazolium bromide ( MTT) assay. Cell apoptosis induced by MALAT1 overexpression, knockdown of ERBB3 and／or osimertinib treatment were analyzed by flow cytometry analysis. The expressions of EGFR and ERBB3 signal pathway related proteins in HCC827 cells treated with overexpression of MALAT1, knockdown of ERBB3 and／or osimertinib treatment were detected by western blot. Results The MTT assay showed that sensitivity to osimertinib of HCC827／MALAT1 cells were significantly repressed. The 50% inhibitive concentration (IC50 ) of osimertinib ＞4 000 nmol／L in HCC827／MALAT1 cells.However, knockdown of ERBB3 facilitated the anti?proliferation effect of osimertinib, and the IC50 of osimertinib in shRNA?ERBB3 cells was (17.27±3.21) nmol／L. The results of flow cytometry analysis showed that the apoptotic rate of HCC827／MALAT1 cells induced by 10 nmol／L osimertinib was (8.38±0.92)%, significantly lower than ( 27.17± 5.83)% of knockdown of ERBB3 ( P＜0.01). Western blotting showed that the expression of p?ERBB3, p?AKT and p?extracellular regulated protein kinases ( ERK) in HCC827／MALAT1 cells was markedly up?regulated, while the expression of p?epithelial growth factor receptor (EGFR) was inhibited. The expressions of p?ERBB3, p?AKT and p?ERK were marginally affected by osimertinb. However, osimertinib downregulated the expressions of p?EGFR, p?ERBB3, p?AKT and p?ERK in ERBB3 deleted cells. Conclusions MALAT1 confers resistance to osimertinb in HCC827 cells by activating of the ERBB3／PI3K／AKT and ERBB3／MAPK／ERK signaling pathways.
