Expression and functional activity of heat shock proteins in human glioblastoma multiforme

Objective: To assess the expression of heat shock proteins (HSP) in glioma cells in vitro and in vivo and to examine their role in resistance to apoptosis. Background: HSP are expressed in response to various forms of stress. Constitutive HSP expression may confer resistance to cytotoxic stimuli in human cancers. Methods: HSP expression was assessed by immunoblot analysis in glioma cells in vitro and by immunocytochemistry in human glioblastomas in vivo. Modulation of apoptosis by hyperthermia-mediated HSP induction was examined in glioma cell lines in vitro. Results: Immunoblot analysis revealed constitutive expression of HSP27, HSP72, HSP73, and HSP90 in all 12 human glioma cell lines. αB-crystallin (αBC) was expressed in 3 of 12 cell lines. High levels of αBC and HSP72 correlated with drug resistance and high p53 levels in vitro. Transient hyperthermia (43 °C/2 hours) induced HSP27 and HSP72 expression but had no effect on the levels of αBC, HSP73, or HSP90. HSP induction provided no survival advantage against subsequent cytotoxic challenges, including cytotoxic cytokines and radiochemotherapy. Immunohistochemistry showed strong expression of all HSP in vivo. The comparative analysis of HSP27, αBC, HSP72, HSP73, and HSP90 expression in 24 paired samples of first resections and recurrences of human glioblastoma multiforme revealed no impact of HSP expression on response to adjuvant radiochemotherapy and no modulation of HSP expression by radiochemotherapy. Conclusions: High constitutive, as opposed to inducible, expression of HSP may play a role in the primary resistance of human malignant gliomas to cytotoxic radiochemotherapy. Superinduction of HSP levels by hyperthermia in vitro provided no further survival advantage.

[1]  I. Konishi,et al.  Expression of heat shock proteins HSP70 and HSP90 in endometrial carcinomas: Correlation with clinicopathology, sex steroid receptor status, and p53 protein expression , 1996, Cancer.

[2]  M. Jäättelä,et al.  Major heat shock protein hsp70 protects tumor cells from tumor necrosis factor cytotoxicity. , 1992, The EMBO journal.

[3]  J. Sambrook,et al.  HSP70 Binding Sites in the Tumor Suppressor Protein p53* , 1997, The Journal of Biological Chemistry.

[4]  T. Ishida,et al.  Overexpression of resistance‐related proteins (metallothioneins, glutathione‐S‐transferase π, heat shock protein 27, and lung resistance‐related protein) in osteosarcoma , 1997, Cancer.

[5]  O. Wiestler,et al.  Expression of αB‐crystallin in human brain tumors , 1993 .

[6]  L. Schiaffonati,et al.  Expression of the HSP 70 gene family in rat hepatoma cell lines of different growth rates. , 1991, Experimental cell research.

[7]  T. Ohnishi,et al.  Suppression of heat-induced hsp72 accumulation by cisplatin in human glioblastoma cells. , 1996, Cancer letters.

[8]  M. Quaia,et al.  Expression of heat shock protein 72 in renal cell carcinoma: possible role and prognostic implications in cancer patients. , 1997, European journal of cancer.

[9]  C. Kittas,et al.  Heat shock protein 70 and HLA-DR molecules tissue expression , 1995, Diseases of the colon and rectum.

[10]  E. Ohama,et al.  Comparative study on the expression of stress‐response protein (srp) 72, srp27, αB‐crystallin and ubiquitin in brain tumours. An immunohistochemical investigation , 1993, Neuropathology and applied neurobiology.

[11]  J. Selkirk,et al.  Multiple p53 protein isoforms and formation of oligomeric complexes with heat shock proteins Hsp70 and Hsp90 in the human mammary tumor, T47D, cell line. , 1994, Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society.

[12]  J. Smyth,et al.  Expression of the heat shock protein HSP27 in human ovarian cancer. , 1995, Clinical cancer research : an official journal of the American Association for Cancer Research.

[13]  J. Dichgans,et al.  Predicting chemoresistance in human malignant glioma cells: The role of molecular genetic analyses , 1998, International journal of cancer.

[14]  H. Ford,et al.  Induction of heat shock protein 70 protects thymocytes against radiation-induced apoptosis. , 1997, Archives of surgery.

[15]  L. Neckers,et al.  Mutant conformation of p53 translated in vitro or in vivo requires functional HSP90. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[16]  S. Kaul,et al.  Elevated levels of mortalin expression in human brain tumors. , 1997, Experimental cell research.

[17]  M. Weller,et al.  Anti-Fas/APO-1 antibody-mediated apoptosis of cultured human glioma cells. Induction and modulation of sensitivity by cytokines. , 1994, The Journal of clinical investigation.

[18]  M. Weller,et al.  APO2 ligand: a novel lethal weapon against malignant glioma? , 1998, FEBS letters.

[19]  A. Giaccia,et al.  Heat Shock Protein 72 Modulates Pathways of Stress-induced Apoptosis* , 1998, The Journal of Biological Chemistry.

[20]  L. Weber,et al.  Effect of overexpression of the small heat shock protein HSP27 on the heat and drug sensitivities of human testis tumor cells. , 1996, Cancer research.

[21]  M. Fukui,et al.  Distinctive immunohistochemical profiles of small heat shock proteins (Heat shock protein 27 and αB‐crystallin) in human brain tumors , 1996, Cancer.

[22]  T. Iwaki,et al.  AlphaB-crystallin protects glial cells from hypertonic stress. , 1996, The American journal of physiology.

[23]  G. Sotiropoulou-Bonikou,et al.  Prognostic significance of Hsp-27 in astrocytic brain tumors: an immunohistochemical study. , 1997, Anticancer research.

[24]  K. Schulze-Osthoff,et al.  Small Stress Proteins as Novel Regulators of Apoptosis , 1996, The Journal of Biological Chemistry.

[25]  F. Gago,et al.  Heat shock protein expression and drug resistance in breast cancer patients treated with induction chemotherapy , 1998, International journal of cancer.

[26]  M. Simon,et al.  Drug resistance against gemcitabine and topotecan mediated by constitutive hsp70 overexpression in vitro: implication of quercetin as sensitiser in chemotherapy. , 1996, British Journal of Cancer.

[27]  M. Jäättelä,et al.  Heat shock protects WEHI‐164 target cells from the cytolysis by tumor necrosis factors α and β , 1989 .

[28]  J. Jenkins,et al.  Mutant p53 proteins bind hsp 72/73 cellular heat shock-related proteins in SV40-transformed monkey cells. , 1987, Oncogene.

[29]  T G Cotter,et al.  Heat shock proteins increase resistance to apoptosis. , 1996, Experimental cell research.

[30]  H. Schipper,et al.  Differential effects of cysteamine on heat shock protein induction and cytoplasmic granulation in astrocytes and glioma cells. , 1995, Brain research. Molecular brain research.