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Inhibition of HSP90‐dependent telomerase activity in amyloid β‐induced apoptosis of cerebral endothelial cells
Author(s) -
Chiu WenTa,
Shen ShingChuan,
Yang LiangYo,
Chow JyhMing,
Wu ChinYen,
Chen YenChou
Publication year - 2011
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.22536
Subject(s) - telomerase , hsp90 , geldanamycin , apoptosis , protein kinase b , telomerase reverse transcriptase , microbiology and biotechnology , chemistry , heat shock protein , ly294002 , cytotoxic t cell , biology , biochemistry , gene , in vitro
Although apoptosis induced by amyloid β (Aβ) has been identified, the effect of Aβ on telomerase activity in relation to apoptosis induction remains unclear. In the present study, Aβ 1–40 and Aβ 25–35 , but not Aβ 1–16 and Aβ 35–25 , reduce the viability of primary cerebral endothelial cells (CECs) in accordance with apoptosis induction. Increases in caspase 3 and PARP protein cleavage with reductions of the Bcl‐2/Bax protein ratio accompanied by a loss in the mitochondria membrane potential were identified in Aβ 1–40 and Aβ 25–35 ‐treated CECs. A significant decrease in intracellular telomerase activity by Aβ 1–40 and Aβ 25–35 was detected; meanwhile, reduced telomerase activity by telomerase reverse transcriptase (TERT) siRNA enhanced the cytotoxic effect of Aβ. The addition of serum might block the Aβ 25–35 ‐induced cytotoxic effect via elevated telomerase activity in according with stimulating phospho‐AKT protein expression, which was blocked by adding AKT inhibitor LY294002. Decreases in heat shock protein 90 (HSP90) and its client proteins including TERT, AKT, p53, CDK4 were observed in Aβ 1–40 and Aβ 25–35 , but not Aβ 1–16 and Aβ 35–25 , ‐treated CECs. The knockdown of HSP90 gene expression by HSP90 siRNA significantly inhibits telomerase activity with decreasing TERT protein expression. The application of HSP90 activity inhibitor geldanamycin (GA) and radicicol (RD) potentiates the telomerase inhibition and apoptosis induction of Aβ in CECs. An increase in protein ubiquitination by Aβ 25–35 , but not Aβ 35–25 , treatment was examined, and Aβ‐inhibited HSP90 and TERT protein expression and telomerase activity was reversed by adding proteasome inhibitor, MG132. Additionally, increased TERT protein ubiquitination by Aβ 25–35 was detected in CECs via immunoprecipitation/Western blotting analysis. The data of the present study firstly demonstrates that telomerase inhibition contributes to the apoptosis induction of Aβ in CECs. J. Cell. Physiol. 226: 2041–2051, 2011. © 2010 Wiley‐Liss, Inc.