Decrease of wild type p53 protein might be due to the regulation of HDAC inhibitors on p53 transcription. Peltonen et al. dis covered that TSA stabilized wild type p53 in melanoma cell lines, but p53 protein accumulation was overridden by simultaneous downregulation of p53 mRNA, leading to a decrease in p53 protein. The mechanisms of p53 acetylation on both wild type and mutant proteins selleck chem inhibitor in dif ferent tumors after various HDACi exposure requires fur ther investigation. The Akt pathway plays an important role in cell growth, and its activation is common in tumors. Inhib ition of overphosphorylated Akt is a promising target ther apy in colorectal cancer . We observed pAkt overexpression in all three cell lines and subsequent downregulation after TSA treatment. A similar phenomenon was reported in other studies.

Chen et al. demon strated that HDACi caused Akt dephosphorylation in U87MG glioblastoma and PC 3 prostate cancer cells by disrupting HDAC protein phosphatase 1 complexes. LBH, another HDACi with a chemical structure similar to TSA, mediated Akt dephosphory lation in DLBCL DHL 6 cells through increased bind ing of PP1 to Akt. We further studied the downstream targets in the Akt pathway. Upregulation of p21 was previously commonly reported, with less data on p27. Repression of cyclin D1 from HDAC inhibitors was reported in mantle cell lymph oma. In our study, we found more significant al terations of p27 and cyclin D1 than p21 after TSA treatment. Both p21 and p27 were upregulated, and cyclin D1 was downregulated with decreasing expres sion of pAkt, which may account for the eventual cell cycle delay.

TSA also induces cell apoptosis in LY1 and LY8 cells. Bcl 2, an anti apoptosis regulator, was found to be downregulated after TSA treatment in LY1 and LY8 cells. In normal germinal centers, Bcl 2 is usually inactivated, rendering centroblasts and centrocytes susceptible to apop tosis. Abnormal retention of Bcl 2 leads to cells that do not die, thereby predisposing cells to malignant transformation. In our study, western blot analysis showed that the repres sion of Bcl 2 occurred at the translational level in LY1 and LY8 cells after TSA treatment. Its downregulation may be the combined effect of Akt dephosphorylation and p53 acetylation caused by TSA. However, Bcl 2 alteration in DoHH2 cells was quite different with LY1 and LY8 cells.

Bcl 2 gene rearrangement was previously reported in DoHH2, LY1 and LY8 cells. However, there is no detailed information regarding Bcl 2 amplification in the li terature. Our unpublished data showed that all three cell lines do not have apparent Bcl 2 gene amplification. One reason for the differential effects on Bcl 2 may be due to different levels of p53 acetylation. Low p53 acetylation Dacomitinib may contribute to DoHH2 cells resistance to apoptosis after TSA treatment at IC50.