title: [Overexpression of Keap1 inhibits the cell proliferation and metastasis and overcomes the drug resistance in human lung cancer A549 cells].
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Institute of Cancer Pharmacology, the First Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou 310006, China.
authors.forename: X
authors.initials: X
authors.lastname: Weng
authors.affiliation: Translational Medicine Research Center, the First People's Hospital, Hangzhou 310006, China.
authors.forename: Y Y
authors.initials: YY
authors.lastname: Yan
authors.affiliation: Department of Clinical Pharmacy, Zhejiang Cancer Hospital, Hangzhou 310022, China.
authors.forename: Y H
authors.initials: YH
authors.lastname: Tong
authors.affiliation: Department of Clinical Pharmacy, Zhejiang Cancer Hospital, Hangzhou 310022, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Fan
authors.affiliation: Institute of Cancer Pharmacology, the First Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou 310006, China.
authors.forename: J M
authors.initials: JM
authors.lastname: Zeng
authors.affiliation: Institute of Cancer Pharmacology, the First Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou 310006, China.
authors.forename: L L
authors.initials: LL
authors.lastname: Wang
authors.affiliation: Translational Medicine Research Center, the First People's Hospital, Hangzhou 310006, China.
authors.forename: N M
authors.initials: NM
authors.lastname: Lin
mesh_terms: D000072283:A549 Cells; D000970:Antineoplastic Agents / Q000494:pharmacology*; D016190:Carboplatin / Q000494:pharmacology; D049109:Cell Proliferation; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000494:pharmacology; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D000072019:Kelch-Like ECH-Associated Protein 1 / Q000235:genetics / Q000378:metabolism*; D008175:Lung Neoplasms / Q000235:genetics / Q000378:metabolism*; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D000093542:Gemcitabine
chemical_list: D000970:Antineoplastic Agents; C431364:KEAP1 protein, human; D000072019:Kelch-Like ECH-Associated Protein 1; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D003841:Deoxycytidine; D016190:Carboplatin; D000093542:Gemcitabine