In this study, ATL-962 PXD101 inhibited these pathways in the sensitive cell line 8505C, but not in WRO82-1 and BHP7-13. The data imply that the ability of PXD101 to inhibit RAS/RAF/ERK and PI3K/AKT/ mTOR pathways may confer sensitivity. p-H2AX is a classic marker of DSBs, a serious type of DNA damage. PXD101 significantly induced p-H2AX in three thyroid cancer cell lines, supporting DSBs as one mechanism accounting for the cytotoxicity of PXD101. We show that PXD101 decreases DSBs repair proteins in the NHEJ and HR pathways. For NHEJ, the KU70-KU80 heterodimer recognizes DSBs, and recruits a DNA protein MEDChem Express Natural Black 1 kinase complex. The decreased expression of KU70 or KU80 can impair the NHEJ repair pathway. For HR, RAD51 binds to the resected end of single-stranded DNA, allowing for synthesisdependent strand annealing for DSB repair. An analysis of the extent of the contribution of individual HR proteins to DNA repair and found that depletion of RAD51 induces the most significant HR defect. The SSA pathway is an alternative mechanism for DSBs repair when NHEJ and HR are defective. PXD101 increased RAD52 and ERCC1, suggesting the SSA pathway was activated. However, when p-H2AX is increased, activation of SSA seems insufficient to repair DSBs. In addition to PXD101, other HDAC inhibitors are able to repress DSB repair proteins. Some HDACs are responsible for DNA repair; HDAC1 and 2 promote NHEJ, and HDAC 9 and 10 are required for HR. PXD101 treatment significantly repressed 8505C tumor growth during the study period. PXD101 transiently increased acetylation of histone H4 that is consistent with prior report. Increase of p-H2AX suggests PXD101 induced DSBs in 8505C xenografts. The anti-tumor effect of PXD101 may be through apoptosis and inhibition of proliferation since caspase-3 and PCNA were decreased. No significant weight loss or toxicity observed in this study, suggesting a favorable safety profile. We also examined the therapeutic effects of PXD101 in mice bearing TT tumors. Daily intraperitoneal injection of PXD101 for 5 doses per week failed to repress the growth of TT xenografts. It is possible that a more intensive PXD101 treatment regimen may affect the growth of TT xenografts. ATC is the most aggressive thyroid cancer and is typically fatal, with a 1-year s
