title: [In vitro construction and confirmation of adenovirus containing a mifepristone (RU486)-inducible regulation system for NRF2 gene].
abstract: OBJECTIVE
  To construct an adenovirus containing a mifepristone (RU486)-inducible regulation system for NRF2 gene, express the product in H460 cell and verify whether the mentioned system can control the gene expression and assess its efficiency.
  
  
  METHODS
  A RU486-inducible regulation system for Nrf2 gene was introduced into an adenovirus. The confirmation was performed through the LUC and Dsred genes. And the expression pattern of Nrf2 at the viral level was examined by Western blot and RT-PCR (reverse transcription-polymerase chain reaction).
  
  
  RESULTS
  The expressions of LUC and Dsred showed a rising trend with the incremental dose of RU486. After the transfection H460 cell with Ad-RUNrf2, the results of RT-PCR and Western blot demonstrated that the expression of Nrf2 was elevated with a rising dose of RU486. After the removal of RU486, the expression of Nrf2 was reduced.
  
  
  CONCLUSIONS
  The construction of an adenovirus carrying Nrf2 gene regulated by a RU486-inducible system is successful, and RU486 can adjust the cellular expression of Nrf2 factor. The LUC and the Dsred expression assumes the dosage dependence along with RU486 to increase; after the Ad-RUNrf2 infects the H460 cell, through RTPCR and Western the Blot result demonstrated that the expression of Nrf2 increases along with the RU486 dosage increases, after removing RU486, the Nrf2 expression is weaken. Showing the construction of the adenovirus carrying Nrf2 gene regulated by the mifepristone (RU486)-inducible system is successful, and RU486 can adjust the Nrf2 factor in the cell the expression.
authors.affiliation: Emergency Medical Department, First Affiliated Hospital, Wenzhou Medical College, Wenzhou 325000, China.
authors.forename: Qi-qi
authors.initials: QQ
authors.lastname: Cai
authors.forename: Guang-liang
authors.initials: GL
authors.lastname: Hong
authors.forename: Qiao-meng
authors.initials: QM
authors.lastname: Qiu
authors.forename: Lin-fang
authors.initials: LF
authors.lastname: Li
authors.forename: Zhong-qiu
authors.initials: ZQ
authors.lastname: Lu
authors.forename: Hong-ping
authors.initials: HP
authors.lastname: Wu
authors.forename: Duan-ming
authors.initials: DM
authors.lastname: Wang
authors.forename: Huan
authors.initials: H
authors.lastname: Liang
mesh_terms: D000256:Adenoviridae / Q000187:drug effects / Q000235:genetics*; D002460:Cell Line; D015870:Gene Expression; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D015735:Mifepristone / Q000378:metabolism / Q000494:pharmacology*; D051267:NF-E2-Related Factor 2 / Q000235:genetics*; D011401:Promoter Regions, Genetic; D014162:Transfection
chemical_list: D051267:NF-E2-Related Factor 2; D015735:Mifepristone