The Subcellular Localization Of DJ-1in The Human Leukemia HL-60Cells Induced By DADS

Objective:To investigate the subcellular localization of DJ^-1 induced by DADS in humanleukemic cell line HL-60cell, and to explore the molecular mechanisms of humanleukemic cell line induced by DADS, provide an important basis for studying thefunction of DJ^-1 protein.Method：HL-60cells induced by DADS were collected at different time, the cytoplasm,nucleus and mitochondria of cells were isolated by cellular components isolation kit,the expression of DJ^-1 were detected by western blotting. The subcellular localizationof DJ^-1 in HL-60cell induced by DADS was detected by immunocytochemistry andimmunofluorescence.Results：1. The expression of DJ^-1 protein in the cytoplasm and nucleus of HL-60cellsinduced by DADS.Western blot results showed that the expression of cytoplasm DJ^-1 protein inHL-60cells group which was treated with DADS1.25mg·L^-1for8h was significantlyreduced （P<0.05）, while the expression of DJ^-1 protein in cell nucleus wassignificantly increased （P<0.05）. The results of Immunofluorescence andimmunocytochemistry confirmed that the expression of DJ^-1 in the cytoplasm wasincreased while decreased in the nucleus after treated with DADS8h in HL-60,showed that DJ^-1 will transfer from the nucleus to the cytoplasm after8h. HL-60cellswere incubated with retinoic acid at different concentrations (2.5μmol·L^-1,5.0μmol·L^-1,10μmol·L^-1).The expression of DJ^-1 in cytoplasm of HL-60cells in5.0μmol·L^-1,10.0μmol·L^-1groups comparing to control group （P<0.01）, while theexpression of DJ^-1 in nucleus of HL-60cells had no significant difference. 2. The expression of DJ^-1 protein in mitochondria of HL-60cells induced byDADS.Western blot results showed that the expression of DJ^-1 protein in themitochondria of HL-60cells treated by DADS1.25mg·L^-1for0,4,8and12h had nodifferences （P>0.05）, but while treated HL-60cells for8h in different concentration(1.25mg·L^-1,2.5mg·L^-1,5.0mg·L^-1,10.0mg·L^-1), the expression of DJ^-1 protein inmitochondria at5.0mg·L^-1,10.0mg·L^-1groups were decreased significantly（P<0.05）.Conclusion：The expression of DJ^-1 in the cytoplasm was increased while decreased in thenucleus of HL-60cells treated with DADS after8h, showed that DJ^-1 protein willtransfer from the nucleus to the cytoplasm.The expression of DJ^-1 in mitochondria of HL-60cells was decreased whentreated with5.0mg·L^-1and10.0mg·L^-1DADS.