It is well known that Erk regulate STAT3 activity negatively. Erk activity may partially contribute to everolimus induced cell growth inhibition in keratinocyte. p38 MAPK pathways are known as stress response signals and interact with the PI3K Akt mTOR pathway. Recently, it was reported that kera tinocyte apoptosis induced by gefitinib, which is a selective EGFR tyrosine kinase inhibitor, is mediated by the JNK activation pathway. This study did not reproduce the results of that report, therefore, the mechanisms under lying everolimus induced keratinocyte apoptosis may differ from those underlying gefitinib induced apoptosis. Alter natively, they may be one of the gefitinib induced mecha nisms because the gefitinib target signal lies upstream from the target of everolimus.

In addition, because STAT3 Y705F enhanced cell toxicity in HaCaT cells and STAT3C relived, the survival of this type of keratinocytes may depend largely on STAT3. For comparison, we considered that an active form of STAT3 selleck chemicals subtly rescued everolimus induced toxicity because cell temporary transfection efficiency of pcDNA3 STAT3C with lipofection method in HaCaT cells was not higher as a result of confirming STAT3 expressions with western blotting assay. To corroborate this effects of rescue by STAT3C, its necessary in the future to conduct an experiments with HaCaT cells stably expressed STAT3C. Previous reports have suggested that STAT3 inhibition in cutaneous squamous cell carcinoma induces senescence and not apoptosis.

Though apoptosis suppressing kinase inhibitor WIKI4 genes and senescence factors were not evaluated in our study, both apoptotic and senescent effects may have affected the cell growth inhibition in duced by everolimus and the STAT3 inhibitor. In addition, the apoptotic effects observed in our study may have been enhanced by interaction with the effects of mTOR and STAT3 inhibition. Everolimus is distributed by P glycoproteins and me tabolized by CYP3A4. Although the pharmacoki netic profiles of stattic have not been clarified, there is no denying that the interactions between everolimus and stattic are due to pharmacokinetic actions. We have pre viously demonstrated that calcium antagonists and adrenoceptor antagonists enhanced cellular sensitivity to SN 38, an active metabolite of irinotecan, by increasing the concentration of SN 38 in cells.

It is difficult to assume that a similar phenomenon caused the effects observed in this study, however, the involvement of STAT3 may be the greater part of this interaction be cause a similar phenomenon was caused by STA 21, which has a chemical structure that is different from that of stattic, and STAT3C transfection moderated everolimus induced cell growth inhibition. In clinical practice, it is known that the efficacy of mo lecular target drugs is correlated with their toxicity.