SIRT inhibitors induce cell death and p53 acetylation through targeting both SIRT1 and SIRT2.
SIRT proteins play an important role in the survival and drug resistance of tumor cells, especially during chemotherapy. In this study, we investigated the potency, specificity, and cellular targets of three SIRT inhibitors, Sirtinol, Salermide, and EX527. Cell proliferative and cell cycle analyses showed that Sirtinol and Salermide, but not EX527, were effective in inducing cell death at concentrations of 50 micromol/L or over in MCF-7 cells. Instead, EX527 caused cell cycle arrest at G(1) at comparable concentrations. In vitro SIRT assays using a p53 peptide substrate showed that all three compounds are potent SIRT1/2 inhibitors, with EX527 having the highest inhibitory activity for SIRT1. Computational docking analysis showed that Sirtinol and Salermide have high degrees of selectivity for SIRT1/2, whereas EX527 has high specificity for SIRT1 but not SIRT2. Consistently, Sirtinol and Salermide, but not EX527, treatment resulted in the in vivo acetylation of the SIRT1/2 target p53 and SIRT2 target tubulin in MCF-7 cells, suggesting that EX527 is ineffective in inhibiting SIRT2 and that p53 mediates the cytotoxic function of Sirtinol and Salermide. Studies using breast carcinoma cell lines and p53-deficient mouse fibroblasts confirmed that p53 is essential for the Sirtinol and Salermide-induced apoptosis. Further, we showed using small interfering RNA that silencing both SIRTs, but not SIRT1 and SIRT2 individually, can induce cell death in MCF-7 cells. Together, our results identify the specificity and cellular targets of these novel inhibitors and suggest that SIRT inhibitors require combined targeting of both SIRT1 and SIRT2 to induce p53 acetylation and cell death. Mol Cancer Ther; 9(4); 844-55. (c)2010 AACR.
Peck
Barrie
B
Cancer Research-UK Laboratory, Department of Surgery and Cancer, MRC Cyclotron Building, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London W12 0NN, UK.
Chen
Chun-Yuan
CY
Ho
Ka-Kei
KK
Di Fruscia
Paolo
P
Myatt
Stephen S
SS
Coombes
R Charles
RC
Fuchter
Matthew J
MJ
Hsiao
Chwan-Deng
CD
Lam
Eric W-F
EW
D000107:Acetylation / Q000187:drug effects; D001549:Benzamides / Q000494:pharmacology*; D002227:Carbazoles / Q000494:pharmacology*; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004354:Drug Screening Assays, Antitumor; D016193:G1 Phase / Q000187:drug effects; D020868:Gene Silencing / Q000187:drug effects; D006801:Humans; D008239:Lysine / Q000378:metabolism; D008958:Models, Molecular; D009284:Naphthols / Q000494:pharmacology*; D017239:Paclitaxel / Q000494:pharmacology; D010666:Phenylpropionates / Q000494:pharmacology*; D017433:Protein Structure, Secondary; D056564:Sirtuin 1 / Q000737:chemistry / Q000378:metabolism*; D056565:Sirtuin 2 / Q000737:chemistry / Q000378:metabolism*; D014404:Tubulin / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
C550547:6-chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide; D001549:Benzamides; D002227:Carbazoles; C539427:N-(3-((2-hydroxynaphthalen-1-ylmethylene)amino)phenyl)-2-phenylpropionamide; D009284:Naphthols; D010666:Phenylpropionates; D014404:Tubulin; D016159:Tumor Suppressor Protein p53; C439060:sirtinol; C447939:SIRT1 protein, human; C474199:SIRT2 protein, human; D056564:Sirtuin 1; D056565:Sirtuin 2; D008239:Lysine; D017239:Paclitaxel