TRAF6 Targeted Anti-tumor Drug Screening And Mechanism Study

Tumor necrosis factor receptor-associated factor 6(TRAF6)is a member of the TRAF family,it is not only overexpressed in many cancer tissues but also closely associates with tumor cell proliferation,migration and apoptosis.All of the study indicate that TRAF6 play an important role in tumor.We employ computer drug design to find out the compound which could bind with the RING finger doming and first Zn finger domain of TRAF6.Thus,the compound would block the interaction between TRAF6 and Ubc13,which is important for the activation of the downstream pathway.We evaluated effects of N6-Isopentenyladenosine on cervical cancer cell line He La using MTT and flow cytometry.In addition,suppression of the anti-apoptotic protein Bcl-2 and elevation of the pro-apoptotic protein Bax were also observed.We examined effects of N6-Isopentenyladenosine on activations of TRAF6-mediated downstream targets using Western blot or immunoprecipitation.N6-Isopentenyladenosine could reduce He La cell proliferations through apoptosis,and such anti-cancer activity is associated with inhibitions of both AKT and TAK1 signaling pathways.Using HeLa cells,we pursued to determine the effect of N6-Isopentenyladenosine on the activation of AKT and TAK1.Our results showed that N6-Isopentenyladenosine dimished the effect of AKT ubiquitination and AKT phosphorylation at Thr-308 and Ser-473.Similarly,N6-Isopentenyladenosine also attenuated the phosphorylation level of TAK1 at all time points.Anti-proliferation properties of N6-Isopentenyladenosine are likley due to its binding at the RING finger domain of TRAF6 and loss of AKT and TAK1 actitvities as a result of functional modulations of TRAF6.These results support the emerging notion that TRAF6 could serve as a viable target for developing new cancer therapeutics.