Effect of autophagy inhibitor combined with EGFR inhibitor on triple-negative breast cancer MDA-MB-468 and MDA-MB-231 cells
10.3760/cma.j.issn.0253-3766.2016.06.004
- VernacularTitle:自噬抑制剂可增强三阴性乳腺癌细胞系MDA-MB-468和MDA-MB-231对吉非替尼的敏感性
- Author:
Zhaoyun LIU
1
;
Kewen HE
;
Xingguo SONG
;
Xinzhao WANG
;
Peiying ZHUO
;
Xingwu WANG
;
Qinghua MA
;
Zhijun HUO
;
Zhiyong YU
Author Information
1. 250022,济南大学山东省医学科学院医学与生命科学学院
- Keywords:
Breast neoplasms,triple-negative;
Epidermal growth factor receptor;
Gefitinib;
Autophagy inhibitor;
MDA-MB-468 cells;
MDA-MB-231 cells
- From:
Chinese Journal of Oncology
2016;38(6):417-424
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of combined administration of autophagy inhibitor 3?methyladenine/bafilomycin A1 and EGFR inhibitor gefitinib on triple?negative breast cancer MDA?MB?468, MDA?MB?231 cells and estrogen receptor?positive MCF?7 cells. Methods All the cells were treated with 3?methyladenine/bafilomycin A1 and/or gefitinib. The effect of autophagy inhibitor and gefitinib on the cell growth was evaluated by MTT assay. Cell apoptosis was detected by flow cytometry. Western blot analysis was used to determine the alteration of autophagy?related protein ( such as LC3) and apoptosis?related proteins ( such as caspase?3 and caspase?9) . Results MTT assay showed that the IC50 in the GE+3?MA and GE+BAF groups were (4.1±0.2) μmol/L and (3.8±0.3) μmol/L, significantly lower than that of the gefitinib alone group [(7.0±0.2) μmol/L] in MDA?MB?468 cells (P<0.05). Similarly, the IC50 in the GE+3?MA and GE+BAF groups were (9.7±0.1) μmol/L and (7.7±0.2) μmol/L, significantly lower than that of the gefitinib alone group [(14.7±0.1) μmol/L]in MDA?MB231 cells (P<0.05). The flow cytometry assay revealed that the apoptosis rates of MDA?MB?468 cells in GE, GE+3?MA and GE+BAF groups were (12.43± 3.18)%, (23.37±2.71)% and (18.71±2.81)%, respectively. The apoptosis rates of MDA?MB?231 cells of the GE, GE+3?MA and GE+BAF groups were (12.15±1.82)%, (16.94±2.19)% and (33.83±5.92) %, significantly higher than that of the gefitinib alone group (All P<0.05). The apoptosis rates of the MCF?7 cells were not changed significantly among the three groups (P>0.05). Western blot data showed that the expression levels of LC3 and p?Akt were decreased in the combined groups than that of the gefitinib alone group, while the p?PTEN, caspase?3 and caspase?9 were increased. Conclusions Autophagy inhibitor may enhance the sensitivity to gefitinib in MDA?MB?468 and MDA?MB?231 cells by activation of the PTEN/P13K/Akt pathway. Apoptosis in MDA?MB?468 and MDA?MB?231 cells might be enhanced by the combination treatment through caspase cascade.
