Impact of HSP90 Inhibition on Viability and Cell Cycle in Relation to p53 Status

Česká verzia

Authors: M. Pastorek; P. Müller; B. Vojtěšek
Authors place of work: Regionální centrum aplikované molekulární onkologie, Masarykův onkologický ústav, Brno
Published in the journal: Klin Onkol 2016; 29(Supplementum 4): 40-45
Category: Original Articles
doi: https://doi.org/10.14735/amko20164S40

Summary

Background:
Chaperone system inhibition is a recent promising strategy for cancer treatment that exploits increased metabolic needs required for rapid proliferation as well as higher level of proteotoxic stress in neoplastic cells. Chaperone HSP90 plays a key role in proper folding of many de novo synthesized proteins, so-called clients, including tumor suppressor p53 which is commonly mutated in majority of cancers. Aim of this work was therefore to understand the impact of HSP90 inhibition by NVP-AUY922 on breast cancer cell lines with wild-type and mutated p53.

Methods:
Flow cytometry was used to analyze cell viability by fluorescein diacetate assay and changes in cell cycle. Western blotting was used to analyze expression of p53 and p21 proteins.

Results:
Analysis of cell viability after HSP90 inhibition revealed higher sensitivity of cell line with wild-type p53. Cell cycle analysis then showed that both cell lines undergo increase in G2/M block of the cell cycle, but wild-type cell line had also substantial decrease in proliferative capacity of treated cells. We also observed increased expression of negative cell cycle regulator p21 in cell line with wild-type p53.

Conclusions:
Since p21 is directly regulated by p53, our results suggest that mutation status of p53 can be important factor in treatment of breast cancer cells by HSP90 chaperone inhibition and that wild-type p53 can increase sensitivity to HSP90 inhibition.

Key words:
breast cancer – cell cycle – chaperone – HSP90 – TP53 – p21 – p53 – NVP-AUY922

This work was supported by the project MEYS – NPS I – LO1413.

The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study.

The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers.

Submitted:
6. 5. 2016

Accepted:
17. 5. 2016

Zdroje

1. Nathan DF, Vos MH, Lindquist S. In vivo functions of the Saccharomyces cerevisiae Hsp90 chaperone. PNAS 1997; 94 (24): 12949–12956.

2. Echeverria PC, Bernthaler A, Dupuis P et al. An interaction network predicted from public data as a discovery tool: Application to the Hsp90 molecular chaperone machine. PLoS One 2011; 6 (10): e26044. doi: 10.1371/journal.pone.0026044.

3. Taipale M, Krykbaeva I, Koeva M et al. Quantitative analysis of HSP90-client interactions reveals principles of substrate recognition. Cell 2012; 150 (5): 987–1001. doi: 10.1016/j.cell.2012.06.047.

4. Pearl LH, Prodromou C, Workman P. The Hsp90 molecular chaperone: an open and shut case for treatment. Biochem J 2008; 410 (3): 439–453. doi: 10.1042/BJ20071640.

5. Workman P, Burrows F, Neckers L et al. Drugging the cancer chaperone HSP90: combinatorial therapeutic exploitation of oncogene addiction and tumor stress. Ann N Y Acad Sci 2007; 1113: 202–216.

6. Trepel J, Mollapour M, Giaccone G et al. Targeting the dynamic HSP90 complex in cancer. Nat Rev Cancer 2010; 10 (8): 537–549.

7. Parimi S, Tsang RY. Hsp90 inhibitors in oncology: ready for prime time? Curr Oncol 2014; 21 (5): e663–e667. doi: 10.3747/co.21.2163.

8. Miyata Y, Nakamoto H, Neckers L. The therapeutic target Hsp90 and cancer hallmarks. Curr Pharm Des 2013; 19 (3): 347–365.

9. Mimnaugh EG, Chavany C, Neckers L. Polyubiquitination and proteasomal degradation of thep185c-erbB-2 receptor protein-tyrosine kinase induced by geldanamycin. J Biol Chem 1996; 271 (37): 22796–22801.

10. Chavany C, Mimnaugh E, Miller P et al. p185erbB2 binds toGRP94 in vivo. Dissociation of the p185erbB2/ GRP94 heterocomplex by benzoquinoneansamycins precedes depletion of p185erbB2. J Biol Chem 1996; 271 (9): 4974–4977.

11. Vartholomaiou E, Echeverría PC, Picard D. Unusual suspects in the twilight zone between the Hsp90 interactome and carcinogenesis. Adv Cancer Res 2016; 129: 1–30. doi: 10.1016/bs.acr.2015.08.001.

12. Fridman JS, Lowe SW. Control of apoptosis by p53. Oncogene 2003; 22 (56): 9030–9040.

13. Attardi LD, Lowe SW, Brugarolas J et al. Transcriptional activation by p53, but not induction of the p21 gene, is essential for oncogene-mediated apoptosis. EMBO J 1996; 15 (14): 3693–3701.

14. Chen X, Ko LJ, Jayaraman L et al. p53 levels, functional domains, and DNA damage determine the extent of the apoptotic response of tumor cells. Genes Dev 1996; 10 (19): 2438–2451.

15. el-Deiry WS. Regulation of p53 downstream genes. Semin Cancer Biol 1998; 8 (5): 345–357.

16. Sax JK, el-Deiry WS. p53 downstream targets and chemosensitivity. Cell Death Differ 2003; 10 (4): 413–417.

17. Yu J, Zhang L, Hwang PM et al. Identification and classification of p53-regulated genes. PNAS 1999; 96 (25): 14517–14522.

18. Müller L, Schaupp A, Walerych D et al. Hsp90 regulates the activity of wild type p53 under physiological and elevated temperatures. J Biol Chem 2004; 279 (47): 48846–48854.

19. Bell S, Klein C, Müller L et al. p53 contains large unstructured regions in its native state. J Mol Biol 2002; 322 (5): 917–927.

20. Dawson R, Müller L, Dehner A et al. The N-terminal domain of p53 is natively unfolded. J Mol Biol 2003; 332 (5): 1131–1141.

21. Walerych D, Kudla K, Gutkowska M et al. Hsp90 chaperones wild-type p53 tumor suppressor protein. J Biol Chem 2004; 279 (47): 48836–44845.

22. Burrows F, Zhang H, Kamal A. Hsp90 activation and cell cycle regulation. Cell Cycle 2004; 3 (12): 1530–1536.

23. Munoz MJ, Jimenez J. Genetic interactions between Hsp90 and the Cdc2 mitotic machinery in the fission yeast Schizosaccharomyces pombe. Mol Gen Genet 1999; 261 (2): 242–250.

24. Aligue R, Akhavan-Niak H, Russell P. A role for Hsp90 in cell cycle control: wee1 tyrosine kinase activity requires interaction with Hsp90. EMBO J 1994; 13 (24): 6099–6106.

25. Goes FS, Martin J. Hsp90 chaperone complexes are required for the activity and stability of yeast protein kinases Mik1, Wee1 and Swe1. Eur J Biochem 2001; 268 (8): 2281–2228.

26. Lin K, Rockliffe N, Johnson GG et al. Hsp90 inhibition has opposing effects on wild-type and mutant p53 and induces p21 expression and cytotoxicity irrespective of p53/ATM status in chronic lymphocytic leukaemia cells. Oncogene 2008; 27 (17): 2445–2455.

27. Regan PL, Jacobs J, Wang G et al. Hsp90 inhibition increases p53 expression and destabilizes MYCN and MYC in neuroblastoma. Int J Oncol 2011; 38 (1): 105–112.