title: [Nrf2 down-regulated cell line H460-N5 with Keap1 over-expression increased sensitivity to anti-cancer drugs].
abstract: To maked a Nrf2 down-regulated cell line by over-expressing Keap1 in H460 cells to study the role of Nrf2 in drug resistance.
  
  
  
  Transfecting H460 cells with mKeap1-pEGFP and screenig for Keap1 expressing clones by Western blotting with antibodies against Nrf2, HO-1, NQO1 and AKR1C. The cell line with Keap1 over-expression was further confirmed by real-time PCR. The cytotoxicity of H460-N5 to anti-cancer drugs was evaluated by MTS assay.
  
  
  
  MTS assay results showed the enhanced cytotoxicity of anticancer drugs (Oxaliplatin, Doxorubicin and Etopside) to the H460 cell line with keap1 overexpression compared to the control cell line. In H460-N0 cells, the IC(50) values of Oxaliplation and Etopside were 93 micromol/L and 100 micromol/L respectively whereas the IC(50) values of the two drugs were 42 micromol/L and 30 micromol/L correspondingly in H460-N5 cells. A Nrf2 down-regulated cell line H460-N5 and a control cell line with GFP over-expression have been identified.Down-regulation of Nrf2 enhanced the cytotoxicity of Oxaliplatin, Doxorubicin and Etopside. The IC(50) value of Doxorubicin to H460-N0 cell was above 3 mg/L, but that to H460-N5 cell was about 2 mg/L.
  
  
  
  A Nrf2 down-regulated cell line H460-N5 and a control cell line with GFP over-expression have been identified. Down-regulation of Nrf2 enhanced the cytotoxicity of Oxaliplatin, Doxorubicin and Etopside.
authors.affiliation: Department of Biochemistry and Genetics, College of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Li-yan
authors.initials: LY
authors.lastname: Qu
authors.forename: Peng
authors.initials: P
authors.lastname: Gao
authors.forename: Hong-yan
authors.initials: HY
authors.lastname: Wang
authors.forename: Xiu-jun
authors.initials: XJ
authors.lastname: Wang
authors.forename: Xiu-wen
authors.initials: XW
authors.lastname: Tang
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D000975:Antioxidants / Q000378:metabolism / Q000494:pharmacology; D002289:Carcinoma, Non-Small-Cell Lung / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D015536:Down-Regulation; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D000072019:Kelch-Like ECH-Associated Protein 1; D008175:Lung Neoplasms / Q000378:metabolism / Q000473:pathology; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D020218:Response Elements / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology*; D014162:Transfection
chemical_list: D000970:Antineoplastic Agents; D000975:Antioxidants; D047908:Intracellular Signaling Peptides and Proteins; C431364:KEAP1 protein, human; D000072019:Kelch-Like ECH-Associated Protein 1; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human