[Effect of tBHQ and sulforaphane on Nrf2-ARE signaling pathway of Caco2 cells].
To investigate the effect of tBHQ and sulforaphane on the protein expression in Nrf2-ARE signaling pathway of Caco2 cells.



Human colorectal carcinoma Caco2 cells were treated with 20 micromol/L tBHQ and 5 micromol/L sulforaphane (SFN) respectively. Real time PCR, Western blotting and immunoflourescence staining (IF) were performed to measure the target gene expression.



Nrf2, AKR1C1 and NQO1 protein expressions were increased time-dependently in Caco2 cells after treatment with tBHQ and SFN. Time-course experiments showed that tBHQ and SFN increased the accumulation of Nrf2, and concomitantly increased the protein levels of AKR1C1 and NQO1. Real-time PCR and Western blotting showed that tBHQ and SFN significantly increased the expression of Nrf2 at 8h after the treatment, and AKR1C1 and NQO1 at 16 h. Confocal microscopy technique showed that Nrf2 accumulated in the nucleus at 6-8 h after treatment with tBHQ. After 1 h treatment with tBHQ the nuclear Nrf2 maintained at elevated level for at least 4 h with tBHQ withdrawn.



tBHQ and SFN induced nuclear accumulation of Nrf2 and activated Nrf2-dependent regulation of ARE-mediated gene expression in Caco2 cells. In addition, the results provide experimental evidence for choosing the dose and frequency of the inducer in cancer chemoprevention study and in developing inhibitors of Nrf2-ARE signaling pathway.
Wu
Xiao-yuan
XY
Department of Biochemistry and Genetics, College of Medicine, Zhejiang University, Hangzhou, China.
Qu
Li-yan
LY
Quan
Kang
K
Jiang
Yan-ling
YL
Tang
Xiu-wen
XW
D016588:Anticarcinogenic Agents / Q000494:pharmacology; D000975:Antioxidants / Q000378:metabolism* / Q000494:pharmacology; D018938:Caco-2 Cells; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D006801:Humans; D006873:Hydroquinones / Q000494:pharmacology*; D017879:Isothiocyanates; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D018384:Oxidative Stress / Q000235:genetics / Q000502:physiology; D020218:Response Elements / Q000502:physiology; D015398:Signal Transduction / Q000187:drug effects*; D013454:Sulfoxides; D013861:Thiocyanates / Q000494:pharmacology*
D016588:Anticarcinogenic Agents; D000975:Antioxidants; D006873:Hydroquinones; D017879:Isothiocyanates; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D013454:Sulfoxides; D013861:Thiocyanates; C018855:2-tert-butylhydroquinone; D000252:Calcium-Transporting ATPases; C016766:sulforaphane