However, after treatment of 5 uM reversine Paclitaxel for 24 hour, AIF was translocated into the nucleus in OC2 cells, indicating that reversine can trigger the non canonical caspase independent cell death. Reversine Inhibitors,Modulators,Libraries also induces autophagy After treatment of reversine, cells showed intracellular vacuoles as shown in Figure 5A, implying that reversine may induce Inhibitors,Modulators,Libraries the autophagic responses. We checked whether reversine had an effect on the level of type I and II LC3. LC3 II increased obviously within 12 hours after reversine treatment both in OC2 and OCSL cells. Interestingly, the pattern of LC3 in OC2 and OCSL cells under normal condition was dif ferent. Even without reversine stimulation, OC2 cells showed endogenous LC3 II expression. Unlike in OC2, the effect of reversine on LC3 II in OCSL cells was more significant.

Co treatment of the autophagy inhibi tor 3 MA decreased reversine induced LC3 II in both cells, especially in OCSL cells. This result indicated that reversine could trigger autophagy and might contribute to the caspase independent pro grammed cell death. This possibility was further con firmed by the effect of reversine in LC3 II aggregation In OCSL cells ectopically expressing Inhibitors,Modulators,Libraries GFP LC3. GFP LC3 showed diffuse pattern but became puncta pattern if treated with reversine. This puncta pat tern was not observed in the control cells harboring GFP plasmid only in the presence of reversine or 3 MA, ruling out the possibility Inhibitors,Modulators,Libraries of non specific effects of the drugs on puncta formation. Moreover, this puncta pat tern was totally abolished if co treatment reversine with autophagy inhibitor 3 MA.

The quantitative Inhibitors,Modulators,Libraries results are shown in Figure 5D. These data confirmed that reversine triggered autophagic flux in OSCC cells. Reversine inhibits Akt/mTOR signaling pathway Previous studies demonstrated that deregulation of PI3K Akt signaling was frequently observed in various cancer cells including OSCC. For example, over activa tion of Akt may enhance malignancy and resistance of cancer cells through anti apoptotic stress. There fore, PI3K Akt pathway became a promising target for cancer therapy recently. In addition, aurora kinases were also proven as an upstream regulator of Akt activ ity in several cancers recently. Since reversine was an inhibitor of aurora kinase, the effect of reversine on Akt activity deserved further investigations.

Baricitinib purchase As expected, reversine notably reduced the phosphorylation of Akt as well as downstream factors, including mTOR complex 1 and p70S6K within 12 hours. On the contrary, although phospho Jnk was affected slightly at later time, reversine showed no significant influence on the activation of MAPKs pathway. These results indicated that reversine selectively down regulated Akt/mTORC1 signaling pathway, resulting in the suppression of cancer cell growth and induction of autophagy.