Hsp90 Inhibitor SNX-7081 Dysregulates Proteins Involved with DNA Repair and Replication and the Cell Cycle in Human Chronic Lymphocytic Leukemia (CLL) Cells

被引:10
作者
Che, Yiping [1 ]
Best, O. Giles [2 ]
Zhong, Ling [3 ]
Kaufman, Kimberley L. [1 ]
Mactier, Swetlana [1 ]
Raftery, Mark [3 ]
Graves, Lee M. [4 ]
Mulligan, Stephen P. [2 ]
Christopherson, Richard I. [1 ]
机构
[1] Univ Sydney, Sch Mol Biosci, Canc Prote Lab, Sydney, NSW 2006, Australia
[2] Royal N Shore Hosp, Kolling Inst Med Res, Northern Blood Res Ctr, St Leonards, NSW 2065, Australia
[3] Univ New S Wales, Bioanalyt Mass Spectrometry Facil, Kensington, NSW 2052, Australia
[4] Univ N Carolina, Dept Pharmacol, Chapel Hill, NC 27599 USA
基金
澳大利亚国家健康与医学研究理事会;
关键词
Hsp90; inhibitor; chronic lymphocytic leukemia; iTRAQ; Western blot; RT-PCR; drug mechanism; HEAT-SHOCK PROTEINS; P53; TUMOR-ANTIGEN; INITIAL THERAPY; UP-REGULATION; CANCER CELLS; FLUDARABINE; COMPLEX; DAMAGE; CHAPERONE; RESISTANCE;
D O I
10.1021/pr301055y
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The proteomic effects of the Hsp90 inhibitor, SNX-7081, have been determined on the p53-mutated B-cell chronic lymphocytic leukemia (CLL) cell line, MEC1. Following SNX-7081 treatment (500 nM, 24 h), Si proteins changed abundance by more than 2-fold (p < 0.05); 7 proteins increased while 44 proteins decreased. Proteins identified as differentially abundant by LC-MS/MS were validated by Western blotting (DDB1, PCNA, MCM2, Hsp90, Hsp70, GRP78, PDIA6, HLA-DR). RT-PCR showed that SNX-7081 unexpectedly modulates a number of these proteins in MEC1 cells at the mRNA level (PCNA, MCM2, Nup155, Hsp7Q GR278, PDIA6, and HLA-DR). Pathway analysis determined that 3 of the differentially abundant proteins (cyclin D1, c-Myc and pRb) were functionally related. p53 levels did not change upon SNX-7081 treatment of p53 wild-type Raji cells or p53-mutated MEC1 and U266 cells, indicating that SNX-7081 has a p53-independent mechanism. The decreases in DDB1, MCM2, c-Myc, and PCNA and increases of pRb and cyclin D1 were confirmed in MEC1, U266, Raji, and p53 null HL60 cells by Western blotting. These data suggest that SNX-7081 arrests the cell cycle and inhibits DNA replication and repair and provides evidence for the mechanism of the observed synergy between Hsp90 inhibitors and drugs that induce DNA strand breaks.
引用
收藏
页码:1710 / 1722
页数:13
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