Carbamoylating chemoresistance induced by cobalt pretreatment in C6 glioma cells

Putative roles of hypoxia-inducible factor-1

Ding I. Yang, Shang Der Chen, Ya Ting Yang, Tzyh Chwen Ju, Jin Ming Xu, Chung Y. Hsu

Research output: Contribution to journalArticle

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Abstract

1. We tested whether pretreatment of reagents known to induce hypoxia-inducible factor-1 (HIF-1) may confer chemoresistance against cytotoxicity of 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) to rat C6 glioma cells. We also studied which cytotoxic mechanism(s) of chloroethylnitrosoureas could be neutralized by cobalt preconditioning. 2. Preconditioning of rat C6 glioma cells with cobalt chloride (300 μM, 2 h) induced HIF-1 binding activity based on electrophoretic mobility shift assay (EMSA). Results from Western blotting confirmed a heightened HIF-1α level upon cobalt chloride exposure (300-400 μM, 2 h). Cobalt chloride (300 μM) pretreatment for 2 h substantially neutralized BCNU toxicity, leading to increases in glioma cell survival based on MTT assay. In addition, pre-exposure of C6 cells with desferrioxamine (DFO; 400 μM, 3 h), an iron chelator known to activate HIF-1, also induced HIF-1 binding and rendered the glioma cells resistant to cytotoxicity of BCNU. 3. Pre-incubation with cobalt chloride abolished the cytotoxicity of several carbamoylating agents including 2-chloroethyl isocyanate and cyclohexyl isocyanate, the respective carbamoylating metabolites of BCNU and 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea. The protective effect of cobalt exposure, however, was not observed when cells were challenged with alkylating agents including temozolomide. 4. Cadmium chloride (50 μM) effectively reversed cobalt-induced HIF-1 activation. Correspondingly, cadmium chloride suppressed carbamoylating chemoresistance mediated by cobalt chloride pretreatment. Furthermore, both double-stranded oligodeoxynucleotide (ODN) decoy with HIF-1 cognate sequence and antisense phosphorothioate ODNs against HIF-1α partially abolished the carbamoylating chemoresistance associated with cobalt preconditioning. 5. Our results suggest that cobalt- or DFO-preconditioning may enhance glioma carbamoylating chemoresistance that is dependent, at least in part, on induction of HIF-1.

Original languageEnglish
Pages (from-to)988-996
Number of pages9
JournalBritish Journal of Pharmacology
Volume141
Issue number6
DOIs
Publication statusPublished - Mar 2004

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Hypoxia-Inducible Factor 1
Cobalt
Glioma
Carmustine
Cadmium Chloride
temozolomide
Isocyanates
Deferoxamine
Oligodeoxyribonucleotides
Alkylating Agents
Electrophoretic Mobility Shift Assay
Chelating Agents
Cell Survival
Iron
Western Blotting
cobaltous chloride

Keywords

  • Alkylation
  • Antisense phosphorothioate oligodeoxynucleotide
  • BCNU
  • Brain tumor
  • Carbamoylation
  • Chemotherapy
  • Oligodeoxynucleotide decoy

ASJC Scopus subject areas

  • Pharmacology

Cite this

Carbamoylating chemoresistance induced by cobalt pretreatment in C6 glioma cells : Putative roles of hypoxia-inducible factor-1. / Yang, Ding I.; Chen, Shang Der; Yang, Ya Ting; Ju, Tzyh Chwen; Xu, Jin Ming; Hsu, Chung Y.

In: British Journal of Pharmacology, Vol. 141, No. 6, 03.2004, p. 988-996.

Research output: Contribution to journalArticle

Yang, Ding I. ; Chen, Shang Der ; Yang, Ya Ting ; Ju, Tzyh Chwen ; Xu, Jin Ming ; Hsu, Chung Y. / Carbamoylating chemoresistance induced by cobalt pretreatment in C6 glioma cells : Putative roles of hypoxia-inducible factor-1. In: British Journal of Pharmacology. 2004 ; Vol. 141, No. 6. pp. 988-996.
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N2 - 1. We tested whether pretreatment of reagents known to induce hypoxia-inducible factor-1 (HIF-1) may confer chemoresistance against cytotoxicity of 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) to rat C6 glioma cells. We also studied which cytotoxic mechanism(s) of chloroethylnitrosoureas could be neutralized by cobalt preconditioning. 2. Preconditioning of rat C6 glioma cells with cobalt chloride (300 μM, 2 h) induced HIF-1 binding activity based on electrophoretic mobility shift assay (EMSA). Results from Western blotting confirmed a heightened HIF-1α level upon cobalt chloride exposure (300-400 μM, 2 h). Cobalt chloride (300 μM) pretreatment for 2 h substantially neutralized BCNU toxicity, leading to increases in glioma cell survival based on MTT assay. In addition, pre-exposure of C6 cells with desferrioxamine (DFO; 400 μM, 3 h), an iron chelator known to activate HIF-1, also induced HIF-1 binding and rendered the glioma cells resistant to cytotoxicity of BCNU. 3. Pre-incubation with cobalt chloride abolished the cytotoxicity of several carbamoylating agents including 2-chloroethyl isocyanate and cyclohexyl isocyanate, the respective carbamoylating metabolites of BCNU and 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea. The protective effect of cobalt exposure, however, was not observed when cells were challenged with alkylating agents including temozolomide. 4. Cadmium chloride (50 μM) effectively reversed cobalt-induced HIF-1 activation. Correspondingly, cadmium chloride suppressed carbamoylating chemoresistance mediated by cobalt chloride pretreatment. Furthermore, both double-stranded oligodeoxynucleotide (ODN) decoy with HIF-1 cognate sequence and antisense phosphorothioate ODNs against HIF-1α partially abolished the carbamoylating chemoresistance associated with cobalt preconditioning. 5. Our results suggest that cobalt- or DFO-preconditioning may enhance glioma carbamoylating chemoresistance that is dependent, at least in part, on induction of HIF-1.

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