Overexpression of Keap1 inhibits the cell proliferation and metastasis and overcomes the drug resistance in human lung cancer A549 cells
10.3760/cma.j.issn.0253-3766.2016.06.002
- VernacularTitle:Kelch样环氧氯丙烷相关蛋白1过表达抑制肺癌A549细胞的增殖和转移并克服其耐药
- Author:
Xu WENG
1
;
Youyou YAN
;
Yinghui TONG
;
Yun FAN
;
Jianmei ZENG
;
Linling WANG
;
Nengming LIN
Author Information
1. 310006浙江中医药大学附属杭州第一医院肿瘤药理研究所
- Keywords:
Lung neoplasms;
A549 cell line;
Keap1;
Nrf2;
Proliferation;
Metastasis;
Drug resistance,neoplasm
- From:
Chinese Journal of Oncology
2016;38(6):404-410
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of Keap1?Nrf2 pathway on cell proliferation, metastasis and drug resistance of human lung cancer A549 cell line. Methods A549?Keap1 cell line, constantly expressing wild type Keap1, was established by lentiviral transfection. Real?time RT?PCR and western blot were used to determine the expression of Nrf2 and its target gene in A549 cells. Sulforhodamine B ( SRB) assay, flow cytometry, colony formation assay, transwell assay, and cell wound?healing assay were performed to explore the effect of wild type Keap1 expression on the proliferation, invasion, migration and drug resistance of A549 cells. Results Over?expressed Keap1 decreased the expression of Nrf2 protein and
the mRNA level of its downstream target genes and inhibited the ability of cell proliferation and clone formation of A549 cells. Keap1 overexpression induced G0/G1 phase arrest. The percentage of A549?Keap1 cells in G0/G1 phase was significantly higher than that of A549?GFP cells (80.2±5.9)% vs. (67.1±0.9%) (P<0.05). Compared with the invasive A549?Keap1 cells (156.33±17.37), the number of invasive A549?GFP cells was significantly higher (306.67±22.19) in a high power field. Keap1 overexpression significantly enhanced the sensitivity of A549 cells to carboplatin and gemcitabine ( P<0.01) . The IC50s of carboplatin in A549?Keap1 and A549?GFP cells were (52.1±3.3) μmol/L and (107.8±12.9) μmol/L, respectively. The IC50s of gemcitabine in A549?Keap1 and A549?GFP cells were (6.8±1.2) μmol/L and (9.9±0.5) μmol/L, respectively. Conclusion Keap1 overexpression significantly inhibits the expression of Nrf2 and its downstream target genes, suppresses tumor cell proliferation and metastasis, and enhances the sensitivity of A549 cells to anticancer drugs.
