Mechanism of PLOD2 induced osimertinib resistance in non-small cell lung cancer HCC827 cells
10.3760/cma.j.cn112152-20190322-00186
- VernacularTitle:赖氨酸羟化酶2诱导肺癌HCC827细胞对奥希替尼耐药的机制研究
- Author:
Xiaohong KANG
1
;
Ke WANG
;
Ying WANG
;
Hongke ZHAO
;
Jiao ZHANG
;
Kelei ZHAO
;
Zhanhui MIAO
;
Zhenye XU
;
Fei CAO
;
Yabin GONG
Author Information
1. 新乡医学院第一附属医院肿瘤科 453100
- Keywords:
Lung neoplasms;
Procollagen-lysine 2-oxoglutarate 5-dioxygenase 2;
Osimertinib resistance;
Epithelial-mesenchymal transition;
FAK signaling pathway
- From:
Chinese Journal of Oncology
2020;42(3):210-215
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of osimertinib on proliferation, migration and invasion of procollagen-lysine 2-oxoglutarate 5-dioxygenase 2 (PLOD2) overexpressing HCC827 cells and explore the potential mechanism of PLOD2 induced osimertinib resistance.Methods:We transfected HCC827 cells with LV-vector and LV-over/PLOD2. The expression of PLOD2 was detected by quantitative real time polymerase chain reaction (qRT-PCR) and western blotting. The effects of osimertinib on the proliferation of HCC827-vector and HCC827-PLOD2 cells were evaluated by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay. The effects of osimertinib on the migration and invasion of HCC827-vector and HCC827-PLOD2 cells were determined by Transwell assays. The expressions of E-cadherin and vimentin in cells were detected by immunofluorescence (IF). The expressions of epithelial-mesenchymal transition (EMT), FAK-PI3K/AKT and MAPK signal pathway related proteins were detected by western blotting.Results:The MTT assay showed that HCC827-PLOD2 cells were hyposensitive to osimertinib. The 50% inhibitory concentration (IC 50) and resistance index of osimertinib for HCC827-PLOD2 cells was over 1 000 nmol/L and over 100, respectively. The result of wound healing assay showed that the migration distance of HCC827-PLOD2 was about (2.13±0.21) fold changes as that of HCC827-vector cells. The result of Transwell assay showed that the numbers of HCC827-PLOD2 passing through the matrix membrane were (212.78±10.43), significantly higher than (101.32±12.52) of HCC827-vector cells ( P<0.01). The result of IF showed that compared with HCC827-vector cells, the expression of E-cadherin was down-regulated while vimentin was up-regulated in HCC827-PLOD2 cells. Osimertinb downregulated E-cadherin and upregulated vimentin expression in HCC827-vector cells but had limited effect in HCC827-PLOD2 cells. The result of western blotting showed that PLOD2 significantly increased vimentin expression level while decreased E-cadherin expression level. Osimertinib inhibited the expression of p-EGFR, but did not affect the expressions of PLOD2, p-FAK, p-AKT, p-ERK, vimentin and E-cadherin in HCC827-PLOD2 cells. Conclusion:PLOD2 confers resistance to osimertinib in HCC827 cells by regulating EMT, FAK-PI3K/AKT and MAPK signal pathways.
