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Degradation of retinoid X receptor α by TPA through proteasome pathway in gastric cancer cells

【摘要】:##正## AIM:To investigate and determine the mechanism and signalpathway of tetradecanoylphorbol-1,3-acetate(TPA)indegradation of RXRα.METHODS:Gastric cancer cell line,BGC-823 was used inthe experiments.The expression level of RXRα protein wasdetected by Western blot.Nuclear and cytoplasmic proteinfractions were prepared through lysis of cell and centrifugation.Localization and translocation of RXRα were observed underlaser-scanning confocal microscope through labeling specificanti-RXRα antibody and corresponding immunofluorescentantibody as secondary antibody.Different inhibitors wereused as required.RESULTS:In BGC-823 cells,RXRα was expressed in thenucleus.When cells were treated with TPA,expression ofRXRα was repressed in a time-dependent and TPA-concentration-dependent manner.Meanwhile,translocationof RXRα from the nucleus to the cytoplasm occurred,also ina time-dependent manner.When cells were pre-incubatedwith proteasome inhibitor MG132 for 3 hrs,followed by TPAfor another 12 hrs,TPA-induced RXRα degradation wasinhibited.Further observation of RXRα translocation in thepresence of MG132 showed that MG-132 could block TPA-induced RXRα redistribution.Conversely,when RXRαtranslocation was inhibited by LMB,an inhibitor for blockingprotein export from the nucleus,TPA could not repressexpression of RXRα.CONCLUSION:TPA could induce the degradation ofRXRα protein in BGC-823 cells,and this degradation istime-and TPA-concentration-dependent.Furthermore,thedegradation of RXRα by TPA is via a proteasome pathwayand associated with RXRα translocation from the nucleusto the cytoplasm.

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