Effect of a heat shock protein 90‑specific inhibitor on the proliferation and apoptosis induced by VEGF‑C in cervical cancer cells

Du,Xue; Li,Yongmei; Jing,Xu; Zhao,Lina

doi:10.3892/etm.2014.1930

Effect of a heat shock protein 90‑specific inhibitor on the proliferation and apoptosis induced by VEGF‑C in cervical cancer cells

Authors:
- Xue Du
- Yongmei Li
- Xu Jing
- Lina Zhao
View Affiliations

Affiliations: Department of Obstetrics and Gynecology, General Hospital, Tianjin Medical University, Tianjin 300052, P.R. China, Department of Microbiology and Immunology, Tianjin Medical University, Tianjin 300052, P.R. China, Department of Biological Engineering, College of Life Sciences, Hebei United University, Tangshan, Hebei 063000, P.R. China
Published online on: August 22, 2014 https://doi.org/10.3892/etm.2014.1930
Pages: 1559-1564

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The aim of the present study was to investigate the effect of heat shock protein 90 (Hsp90)‑specific inhibitor geldanamycin (GA) on the proliferation and apoptosis induced by vascular endothelial growth factor‑C (VEGF‑C) in cervical cancer cells. HeLa cells (1x106/ml) in the logarithmic growth phase were incubated without serum for 24 h. The cells were pretreated with kinase insert domain receptor antibody (KDR)‑Ab (20 µg/ml), phosphatidylinositol 3‑kinase (PI3K) inhibitor LY294002 (3 µmol/l), mitogen‑activated protein kinase (MAPK) inhibitor PD98059 (30 µmol/l) or Hsp90‑specific inhibitor GA (10 µmol/l) for 30 min, and then treated with VEGF‑C (50 ng/µl) for a further 24 h. The cells were harvested for MTT analysis, annexin V‑FITC/propidium iodide double staining for early apoptosis and SDS‑PAGE and western blot analysis in order to determine Hsp90, B‑cell lymphoma 2 (Bcl‑2), Bcl‑2‑associated X protein (Bax) and cyclin D1 expression. Treatment with VEGF‑C alone induced Hsp90 protein expression in HeLa cells at all time‑points. Hsp90 expression was increased 3.31‑fold in VEGF‑C treated HeLa cells, and this increase was attenuated in the treatment groups (2.17‑, 1.69‑, 1.82‑fold in VEGF‑C + KDR‑Ab, VEGF‑C + PD98059 and VEGF‑C + LY294002, respectively). The proliferation of the VEGF‑C‑treated HeLa cells was increased ~2.13‑fold, while that of the VEGF‑C + GA‑treated HeLa cells decreased 0.87‑fold (P<0.05). Even low concentrations of GA (0.02 µmol/l) were found to inhibit the Bcl‑2 and cyclin D1 protein expression induced by VEGF‑C. Therefore, the results indicate that the Hsp90‑specific inhibitor GA has a critical role in the proliferation and apoptosis induced by VEGF‑C in cervical cancer cells.

View Figures

View References

1	Bishop SC, Burlison JA and Blagg BS: Hsp90: a novel target for the disruption of multiple signaling cascades. Curr Cancer Drug Targets. 7:369–388. 2007. View Article : Google Scholar : PubMed/NCBI
2	Trepel J, Mollapour M, Giaccone G and Neckers L: Targeting the dynamic HSP90 complex in cancer. Nat Rev Cancer. 10:537–549. 2010. View Article : Google Scholar : PubMed/NCBI
3	Chiosis G, Dickey CA and Johnson JL: A global view of Hsp90 functions. Nat Struct Mol Biol. 20:1–4. 2013. View Article : Google Scholar : PubMed/NCBI
4	Lee WY, Chen YC, Shih CM, et al: The induction of heme oxygenase-1 suppresses heat shock protein 90 and the proliferation of human breast cancer cells through its byproduct carbon monoxide. Toxicol Appl Pharmacol. 274:55–62. 2014. View Article : Google Scholar : PubMed/NCBI
5	Schwock J, Pham NA, Cao MP and Hedley DW: Efficacy of Hsp90 inhibition for induction of apoptosis and inhibition of growth in cervical carcinoma cells in vitro and in vivo. Cancer Chemother Pharmacol. 61:669–681. 2008. View Article : Google Scholar : PubMed/NCBI
6	Neckers L: Chaperoning oncogenes: Hsp90 as a target of geldanamycin. Handb Exp Pharmacol. 172:259–277. 2006. View Article : Google Scholar : PubMed/NCBI
7	Mehta PP, Kung PP, Yamazaki S, et al: A novel class of specific Hsp90 small molecule inhibitors demonstrate in vitro and in vivo anti-tumor activity in human melanoma cells. Cancer Lett. 300:30–39. 2011. View Article : Google Scholar : PubMed/NCBI
8	Du X and Mi R: Study on the cell signal transductions of cell proliferation and apoptosis induced by VEGF-C in cervical carcinoma. Xian Dai Fu Chan Ke Za Zhi. 11:877–880. 8852011.
9	Chinnaiyan P, Allen GW and Harari PM: Radiation and new molecular agents, part II: targeting HDAC, HSP90, IGF-1R, PI3K, and Ras. Semin Radiat Oncol. 16:59–64. 2006. View Article : Google Scholar : PubMed/NCBI
10	Park JW, Yeh MW, Wong MG, et al: The heat shock protein 90-binding geldanamycin inhibits cancer cell proliferation, down-regulates oncoproteins, and inhibits epidermal growth factor-induced invasion in thyroid cancer cell lines. J Clin Endocrinol Metab. 88:3346–3353. 2003. View Article : Google Scholar
11	Sreedhar AS, Kalmár E, Csermely P and Shen YF: Hsp90 isoforms: functions, expression and clinical importance. FEBS Lett. 562:11–15. 2004. View Article : Google Scholar : PubMed/NCBI
12	Schmitt E, Gehrmann M, Brunet M, Multhoff G and Garrido C: Intracellular and extracellular functions of heat shock proteins: repercussions in cancer therapy. J Leukoc Biol. 81:15–27. 2007. View Article : Google Scholar : PubMed/NCBI
13	Polier S, Samant RS, Clarke PA, et al: ATP-competitive inhibitors block protein kinase recruitment to the Hsp90-Cdc37 system. Nat Chem Biol. 9:307–312. 2013. View Article : Google Scholar : PubMed/NCBI
14	Richter K and Buchner J: Hsp90: chaperoning signal transduction. J Cell Physiol. 188:281–290. 2001. View Article : Google Scholar : PubMed/NCBI
15	Cappello F, David S, Rappa F, et al: The expression of HSP60 and HSP10 in large bowel carcinomas with lymph node metastase. BMC Cancer. 5:1392005. View Article : Google Scholar : PubMed/NCBI
16	Powers MV and Workman P: Targeting of multiple signalling pathways by heat shock protein 90 molecular chaperone inhibitors. Endocr Relat Cancer. 13(Suppl 1): 125–135. 2006. View Article : Google Scholar : PubMed/NCBI
17	Georgakis GV, Li Y, Rassidakis GZ, et al: Inhibition of heat shock protein 90 function by 17-allylamino-17-demethoxy-geldanamycin in Hodgkin’s lymphoma cells down-regulates Akt kinase, dephosphorylates extracellular signal-regulated kinase, and induces cell cycle arrest and cell death. Clin Cancer Res. 12:584–590. 2006.PubMed/NCBI
18	Mitsiades CS, Mitsiades NS, McMullan CJ, et al: Antimyeloma activity of heat shock protein-90 inhibition. Blood. 107:1092–1100. 2006. View Article : Google Scholar : PubMed/NCBI
19	Ferrario A, Rucker N, Wong S, Luna M and Gomer CJ: Survivin, a member of the inhibitor of apoptosis family, is induced by photodynamic therapy and is a target for improving treatment response. Cancer Res. 67:4989–4995. 2007. View Article : Google Scholar : PubMed/NCBI
20	Röhl A, Rohrberg J and Buchner J: The chaperone Hsp90: changing partners for demanding clients. Trends Biochem Sci. 38:253–262. 2013.PubMed/NCBI
21	Sidera K and Patsavoudi E: HSP90 Inhibitors: current development and potential in cancer therapy. Recent Pat Anticancer Drug Discov. 9:1–20. 2014.PubMed/NCBI
22	Gooljarsingh LT, Fernandes C, Yan K, et al: A biochemical rationale for the anticancer effects of Hsp90 inhibitors: slow, tight binding inhibition by geldanamycin and its analogues. Proc Natl Acad Sci USA. 103:7625–7630. 2006. View Article : Google Scholar : PubMed/NCBI
23	Duus J, Bahar HI, Venkataraman G, et al: Analysis of expression of heat shock protein-90 (HSP90) and the effects of HSP90 inhibitor (17-AAG) in multiple myeloma. Leuk Lymphoma. 47:1369–1378. 2006. View Article : Google Scholar : PubMed/NCBI