Dual epidermal growth factor receptor and vascular endothelial growth factor receptor inhibition with vandetanib sensitizes bladder cancer cells to cisplatin in a dose- and sequence-dependent manner

Thomas W. Flaig, Lih Jen Su, Caroline McCoach, Yuan Li, David Raben, Marileila Varella-Garcia, Lynne T. Bemis

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

OBJECTIVE To investigate the activity of the combination of vandetanib and cytotoxic agents using in vitro models of bladder cancer, as modern chemotherapy regimens are built around cisplatin, with gemcitabine or a taxane such as docetaxel also commonly added in combination for the treatment of advanced bladder cancer. MATERIALS AND METHODS Human bladder cancer cells HTB3, HT1376, J82, RT4, CRL1749, T24, SUP and HTB9 were cultured. The activity of gefitinib (ZD1839) and vandetanib (ZD6474) was assessed in these eight bladder cancer cell lines with a tetrazolium-based assay of cell viability. RT4 bladder cancer cells, determined to have moderate cisplatin resistance and also moderate sensitivity to vandetanib, were treated with vandetanib and cisplatin. RT4 and T24 cells were treated with six different regimens. The apoptosis and cell-cycle analysis were studied by flow cytometry. Expression of p21 and p27 was detected by Western blotting. Fluorescence in situ hybridization (FISH) analysis of epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 was performed for all cell lines. RESULTS At equal concentrations, vandetanib was a more potent inhibitor of cell viability, compared to gefitinib. At vandetanib concentrations of ≤2 M, the combination with cisplatin was synergistic, especially in the treatment sequence of cisplatin followed by vandetanib, and additive with vandetanib followed by cisplatin. An analysis of the cell-cycle distribution showed that vandetanib treatment induced G1 arrest at high concentrations, but not at lower concentrations. High-concentration treatment was associated with increased levels of the cyclin-dependent kinase p27. FISH analysis showed that there was a low level of genomic gain, and no gene amplification. Mutational analysis of exons 18, 19, and 21 of EGFR in each cell line revealed no mutation. CONCLUSION Vandetanib has synergistic activity when given at low concentration with cytotoxic chemotherapy. The addition of vandetanib to cisplatin-based chemotherapy regimens merits further study.

Original languageEnglish (US)
Pages (from-to)1729-1737
Number of pages9
JournalBJU International
Volume103
Issue number12
DOIs
StatePublished - Jun 1 2009

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Vascular Endothelial Growth Factor Receptor
Epidermal Growth Factor Receptor
Urinary Bladder Neoplasms
Cisplatin
docetaxel
gemcitabine
Fluorescence In Situ Hybridization
Drug Therapy
Cell Line
N-(4-bromo-2-fluorophenyl)-6-methoxy-7-((1-methylpiperidin-4-yl)methoxy)quinazolin-4-amine
Cell Survival
Cell Cycle
Cyclin-Dependent Kinases
Gene Amplification
Cytotoxins
Therapeutics
Exons
Flow Cytometry
Western Blotting

Keywords

  • Bladder cancer
  • Chemotherapy
  • Epithelial growth factor receptor
  • Vandetanib
  • Vascular endothelial growth factor receptor

Cite this

Dual epidermal growth factor receptor and vascular endothelial growth factor receptor inhibition with vandetanib sensitizes bladder cancer cells to cisplatin in a dose- and sequence-dependent manner. / Flaig, Thomas W.; Su, Lih Jen; McCoach, Caroline; Li, Yuan; Raben, David; Varella-Garcia, Marileila; Bemis, Lynne T.

In: BJU International, Vol. 103, No. 12, 01.06.2009, p. 1729-1737.

Research output: Contribution to journalArticle

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abstract = "OBJECTIVE To investigate the activity of the combination of vandetanib and cytotoxic agents using in vitro models of bladder cancer, as modern chemotherapy regimens are built around cisplatin, with gemcitabine or a taxane such as docetaxel also commonly added in combination for the treatment of advanced bladder cancer. MATERIALS AND METHODS Human bladder cancer cells HTB3, HT1376, J82, RT4, CRL1749, T24, SUP and HTB9 were cultured. The activity of gefitinib (ZD1839) and vandetanib (ZD6474) was assessed in these eight bladder cancer cell lines with a tetrazolium-based assay of cell viability. RT4 bladder cancer cells, determined to have moderate cisplatin resistance and also moderate sensitivity to vandetanib, were treated with vandetanib and cisplatin. RT4 and T24 cells were treated with six different regimens. The apoptosis and cell-cycle analysis were studied by flow cytometry. Expression of p21 and p27 was detected by Western blotting. Fluorescence in situ hybridization (FISH) analysis of epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 was performed for all cell lines. RESULTS At equal concentrations, vandetanib was a more potent inhibitor of cell viability, compared to gefitinib. At vandetanib concentrations of ≤2 M, the combination with cisplatin was synergistic, especially in the treatment sequence of cisplatin followed by vandetanib, and additive with vandetanib followed by cisplatin. An analysis of the cell-cycle distribution showed that vandetanib treatment induced G1 arrest at high concentrations, but not at lower concentrations. High-concentration treatment was associated with increased levels of the cyclin-dependent kinase p27. FISH analysis showed that there was a low level of genomic gain, and no gene amplification. Mutational analysis of exons 18, 19, and 21 of EGFR in each cell line revealed no mutation. CONCLUSION Vandetanib has synergistic activity when given at low concentration with cytotoxic chemotherapy. The addition of vandetanib to cisplatin-based chemotherapy regimens merits further study.",
keywords = "Bladder cancer, Chemotherapy, Epithelial growth factor receptor, Vandetanib, Vascular endothelial growth factor receptor",
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T1 - Dual epidermal growth factor receptor and vascular endothelial growth factor receptor inhibition with vandetanib sensitizes bladder cancer cells to cisplatin in a dose- and sequence-dependent manner

AU - Flaig, Thomas W.

AU - Su, Lih Jen

AU - McCoach, Caroline

AU - Li, Yuan

AU - Raben, David

AU - Varella-Garcia, Marileila

AU - Bemis, Lynne T.

PY - 2009/6/1

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N2 - OBJECTIVE To investigate the activity of the combination of vandetanib and cytotoxic agents using in vitro models of bladder cancer, as modern chemotherapy regimens are built around cisplatin, with gemcitabine or a taxane such as docetaxel also commonly added in combination for the treatment of advanced bladder cancer. MATERIALS AND METHODS Human bladder cancer cells HTB3, HT1376, J82, RT4, CRL1749, T24, SUP and HTB9 were cultured. The activity of gefitinib (ZD1839) and vandetanib (ZD6474) was assessed in these eight bladder cancer cell lines with a tetrazolium-based assay of cell viability. RT4 bladder cancer cells, determined to have moderate cisplatin resistance and also moderate sensitivity to vandetanib, were treated with vandetanib and cisplatin. RT4 and T24 cells were treated with six different regimens. The apoptosis and cell-cycle analysis were studied by flow cytometry. Expression of p21 and p27 was detected by Western blotting. Fluorescence in situ hybridization (FISH) analysis of epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 was performed for all cell lines. RESULTS At equal concentrations, vandetanib was a more potent inhibitor of cell viability, compared to gefitinib. At vandetanib concentrations of ≤2 M, the combination with cisplatin was synergistic, especially in the treatment sequence of cisplatin followed by vandetanib, and additive with vandetanib followed by cisplatin. An analysis of the cell-cycle distribution showed that vandetanib treatment induced G1 arrest at high concentrations, but not at lower concentrations. High-concentration treatment was associated with increased levels of the cyclin-dependent kinase p27. FISH analysis showed that there was a low level of genomic gain, and no gene amplification. Mutational analysis of exons 18, 19, and 21 of EGFR in each cell line revealed no mutation. CONCLUSION Vandetanib has synergistic activity when given at low concentration with cytotoxic chemotherapy. The addition of vandetanib to cisplatin-based chemotherapy regimens merits further study.

AB - OBJECTIVE To investigate the activity of the combination of vandetanib and cytotoxic agents using in vitro models of bladder cancer, as modern chemotherapy regimens are built around cisplatin, with gemcitabine or a taxane such as docetaxel also commonly added in combination for the treatment of advanced bladder cancer. MATERIALS AND METHODS Human bladder cancer cells HTB3, HT1376, J82, RT4, CRL1749, T24, SUP and HTB9 were cultured. The activity of gefitinib (ZD1839) and vandetanib (ZD6474) was assessed in these eight bladder cancer cell lines with a tetrazolium-based assay of cell viability. RT4 bladder cancer cells, determined to have moderate cisplatin resistance and also moderate sensitivity to vandetanib, were treated with vandetanib and cisplatin. RT4 and T24 cells were treated with six different regimens. The apoptosis and cell-cycle analysis were studied by flow cytometry. Expression of p21 and p27 was detected by Western blotting. Fluorescence in situ hybridization (FISH) analysis of epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 was performed for all cell lines. RESULTS At equal concentrations, vandetanib was a more potent inhibitor of cell viability, compared to gefitinib. At vandetanib concentrations of ≤2 M, the combination with cisplatin was synergistic, especially in the treatment sequence of cisplatin followed by vandetanib, and additive with vandetanib followed by cisplatin. An analysis of the cell-cycle distribution showed that vandetanib treatment induced G1 arrest at high concentrations, but not at lower concentrations. High-concentration treatment was associated with increased levels of the cyclin-dependent kinase p27. FISH analysis showed that there was a low level of genomic gain, and no gene amplification. Mutational analysis of exons 18, 19, and 21 of EGFR in each cell line revealed no mutation. CONCLUSION Vandetanib has synergistic activity when given at low concentration with cytotoxic chemotherapy. The addition of vandetanib to cisplatin-based chemotherapy regimens merits further study.

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KW - Chemotherapy

KW - Epithelial growth factor receptor

KW - Vandetanib

KW - Vascular endothelial growth factor receptor

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