The Effect Of Tertiary Butylhydroquinone On Invasion In Hypoxic And Normoxia HepG2 Cell

ObjectivesInvestigate the role and mechanism of tert-butylhydroquinone( t BHQ)in Hep G-2 under different oxygen concentration.MethodsHep G-2 were cultured in vitro,Hypoxia was Modeled using cocl2.MTT analysis measured the OD ration of t BHQ on Hep G-2 and evaluate the cell viability;Western blot analysis measured the effects of tBHQ on the levels of Nrf2 in Hep G-2, DAPI immunofluorescence measured the effects of t BHQ on translocation of Nrf2; Transwell chamber method measured Hep G-2 cell invation and migration capability;Western blot analysis measured the effects of t BHQ on the levels of HIF-1α and smad7. Results1.MTT result showed that t BHQ inhibit cell proliferation under normal oxygen,however promoting proliferation under hypoxia.2.Western Blot showed that the expression of Nrf2 was increased after stimulated t BHQ,compared with control group,the Nrf2 expression of hypoxic group(20umol/L)has no significant difference(p>0.05),compared with control group,the Nrf2 expression of hypoxic group(40umol/L) was increasec(p<0.05);Hypoxia can induce Nrf2 expression.3.DAPI immunofluorescence showd that t BHQ can increse nuclear transfer no matter under hypoxia or normal oxygen and t BHQ can affect Hep G-2 through Nrf2 pathway.4.Transwell chamber method showed that compared with control group, cell invation was higher after t BHQ stimulated under normal oxygen,however,cell invation capability was lower than control group under hypoxia.5. Western Blot showed thatunder normal oxygen the expression of HIF-1α has no significant difference(p>0.05)but stimulated smad7(p< 0.05);under hypoxia the HIF-1α expression was increasec( p < 0.05),smad7 expression was reduced.Conclusions1.TBHQ can inhibit cell invation capability under normal oxygen while promote cell invation capability under hypoxia via inducing translocation of Nrf22.The mechanism may be related to increase HIF- 1α and two-way regulating smad7 expression...