The phosphoinositide 3-kinase/Akt pathway

a new target in human renal cell carcinoma therapy

Carole Sourbier, Véronique Lindner, Hervé Lang, Abdelali Agouni, Eric Schordan, Sabrina Danilin, Sylvie Rothhut, Didier Jacqmin, Jean-Jacques Helwig, Thierry Massfelder

Research output: Contribution to journalArticle

122 Citations (Scopus)

Abstract

Metastatic renal cell carcinoma is resistant to current therapies. The phosphoinositide 3-kinase (PI3K)/Akt signaling cascade induces cell growth, cell transformation, and neovascularization. We evaluated whether targeting this pathway could be of therapeutic value against human renal cell carcinoma. The activation of the PI3K/Akt pathway and its role in renal cell carcinoma progression was evaluated in vitro in seven human cell lines by Western blot, cell counting, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, terminal deoxyribonucleotide transferase-mediated nick-end labeling assays, and fluorescence-activated cell sorting analysis, using two PI3K inhibitors, LY294002 and wortmannin, as well as by transfection with various Akt constructs and through Akt knockdown by small interfering RNA (siRNA). In vivo nude mice bearing human renal cell carcinoma tumor xenografts were treated with LY294002 (75 mg/kg/wk, 4 weeks, i.p.). Tumor growth was measured and tumors were subjected to Western blot and immunohistochemical analysis. Akt was constitutively activated in all cell lines. Constitutive phosphorylation of glycogen synthase kinase-3 (GSK-3) was observed in all cell lines, whereas forkhead transcription factor and mammalian target of rapamycin, although expressed, were not constitutively phosphorylated. Exposure to LY294002 or wortmannin decreased Akt activation and GSK-3 phosphorylation and reduced cell growth by up to 70% through induction of cell apoptosis. These effects were confirmed by transfection experiments with Akt constructs or Akt siRNA. Importantly, LY294002 induced up to 50% tumor regression in mice through tumor cell apoptosis. Tumor neovascularization was significantly increased by LY294002 treatment. Blood chemistries showed no adverse effects of the treatment. Our results suggest an important role of PI3K/Akt inhibitors as a potentially useful treatment for patients with renal cell carcinoma.
Original languageEnglish
Pages (from-to)5130-5142
Number of pages13
JournalCancer Research
Volume66
Issue number10
DOIs
Publication statusPublished - 15 May 2006

Fingerprint

1-Phosphatidylinositol 4-Kinase
Cell- and Tissue-Based Therapy
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Renal Cell Carcinoma
Glycogen Synthase Kinase 3
Neoplasms
Cell Line
Small Interfering RNA
Transfection
Growth
Western Blotting
Phosphorylation
Deoxyribonucleotides
Apoptosis
Forkhead Transcription Factors
Therapeutics
Sirolimus
Transferases
Heterografts
Nude Mice

Keywords

  • 1-Phosphatidylinositol 3-Kinase
  • Animals
  • Carcinoma, Renal Cell
  • Cell Growth Processes
  • Cell Line, Tumor
  • Chromones
  • Enzyme Inhibitors
  • Humans
  • Kidney Neoplasms
  • Male
  • Mice
  • Morpholines
  • Oncogene Protein v-akt
  • Phosphorylation
  • RNA, Small Interfering
  • Signal Transduction
  • Substrate Specificity
  • Transfection
  • Xenograft Model Antitumor Assays

Cite this

Sourbier, C., Lindner, V., Lang, H., Agouni, A., Schordan, E., Danilin, S., ... Massfelder, T. (2006). The phosphoinositide 3-kinase/Akt pathway: a new target in human renal cell carcinoma therapy. Cancer Research, 66(10), 5130-5142. https://doi.org/10.1158/0008-5472.CAN-05-1469

The phosphoinositide 3-kinase/Akt pathway : a new target in human renal cell carcinoma therapy. / Sourbier, Carole; Lindner, Véronique; Lang, Hervé; Agouni, Abdelali; Schordan, Eric; Danilin, Sabrina; Rothhut, Sylvie; Jacqmin, Didier; Helwig, Jean-Jacques; Massfelder, Thierry.

In: Cancer Research, Vol. 66, No. 10, 15.05.2006, p. 5130-5142.

Research output: Contribution to journalArticle

Sourbier, C, Lindner, V, Lang, H, Agouni, A, Schordan, E, Danilin, S, Rothhut, S, Jacqmin, D, Helwig, J-J & Massfelder, T 2006, 'The phosphoinositide 3-kinase/Akt pathway: a new target in human renal cell carcinoma therapy', Cancer Research, vol. 66, no. 10, pp. 5130-5142. https://doi.org/10.1158/0008-5472.CAN-05-1469
Sourbier C, Lindner V, Lang H, Agouni A, Schordan E, Danilin S et al. The phosphoinositide 3-kinase/Akt pathway: a new target in human renal cell carcinoma therapy. Cancer Research. 2006 May 15;66(10):5130-5142. https://doi.org/10.1158/0008-5472.CAN-05-1469
Sourbier, Carole ; Lindner, Véronique ; Lang, Hervé ; Agouni, Abdelali ; Schordan, Eric ; Danilin, Sabrina ; Rothhut, Sylvie ; Jacqmin, Didier ; Helwig, Jean-Jacques ; Massfelder, Thierry. / The phosphoinositide 3-kinase/Akt pathway : a new target in human renal cell carcinoma therapy. In: Cancer Research. 2006 ; Vol. 66, No. 10. pp. 5130-5142.
@article{4ff46a4ea3cb40feb9c3bf8433757814,
title = "The phosphoinositide 3-kinase/Akt pathway: a new target in human renal cell carcinoma therapy",
abstract = "Metastatic renal cell carcinoma is resistant to current therapies. The phosphoinositide 3-kinase (PI3K)/Akt signaling cascade induces cell growth, cell transformation, and neovascularization. We evaluated whether targeting this pathway could be of therapeutic value against human renal cell carcinoma. The activation of the PI3K/Akt pathway and its role in renal cell carcinoma progression was evaluated in vitro in seven human cell lines by Western blot, cell counting, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, terminal deoxyribonucleotide transferase-mediated nick-end labeling assays, and fluorescence-activated cell sorting analysis, using two PI3K inhibitors, LY294002 and wortmannin, as well as by transfection with various Akt constructs and through Akt knockdown by small interfering RNA (siRNA). In vivo nude mice bearing human renal cell carcinoma tumor xenografts were treated with LY294002 (75 mg/kg/wk, 4 weeks, i.p.). Tumor growth was measured and tumors were subjected to Western blot and immunohistochemical analysis. Akt was constitutively activated in all cell lines. Constitutive phosphorylation of glycogen synthase kinase-3 (GSK-3) was observed in all cell lines, whereas forkhead transcription factor and mammalian target of rapamycin, although expressed, were not constitutively phosphorylated. Exposure to LY294002 or wortmannin decreased Akt activation and GSK-3 phosphorylation and reduced cell growth by up to 70{\%} through induction of cell apoptosis. These effects were confirmed by transfection experiments with Akt constructs or Akt siRNA. Importantly, LY294002 induced up to 50{\%} tumor regression in mice through tumor cell apoptosis. Tumor neovascularization was significantly increased by LY294002 treatment. Blood chemistries showed no adverse effects of the treatment. Our results suggest an important role of PI3K/Akt inhibitors as a potentially useful treatment for patients with renal cell carcinoma.",
keywords = "1-Phosphatidylinositol 3-Kinase, Animals, Carcinoma, Renal Cell, Cell Growth Processes, Cell Line, Tumor, Chromones, Enzyme Inhibitors, Humans, Kidney Neoplasms, Male, Mice, Morpholines, Oncogene Protein v-akt, Phosphorylation, RNA, Small Interfering, Signal Transduction, Substrate Specificity, Transfection, Xenograft Model Antitumor Assays",
author = "Carole Sourbier and V{\'e}ronique Lindner and Herv{\'e} Lang and Abdelali Agouni and Eric Schordan and Sabrina Danilin and Sylvie Rothhut and Didier Jacqmin and Jean-Jacques Helwig and Thierry Massfelder",
year = "2006",
month = "5",
day = "15",
doi = "10.1158/0008-5472.CAN-05-1469",
language = "English",
volume = "66",
pages = "5130--5142",
journal = "Cancer Research",
issn = "0008-5472",
publisher = "AMER ASSOC CANCER RESEARCH",
number = "10",

}

TY - JOUR

T1 - The phosphoinositide 3-kinase/Akt pathway

T2 - a new target in human renal cell carcinoma therapy

AU - Sourbier, Carole

AU - Lindner, Véronique

AU - Lang, Hervé

AU - Agouni, Abdelali

AU - Schordan, Eric

AU - Danilin, Sabrina

AU - Rothhut, Sylvie

AU - Jacqmin, Didier

AU - Helwig, Jean-Jacques

AU - Massfelder, Thierry

PY - 2006/5/15

Y1 - 2006/5/15

N2 - Metastatic renal cell carcinoma is resistant to current therapies. The phosphoinositide 3-kinase (PI3K)/Akt signaling cascade induces cell growth, cell transformation, and neovascularization. We evaluated whether targeting this pathway could be of therapeutic value against human renal cell carcinoma. The activation of the PI3K/Akt pathway and its role in renal cell carcinoma progression was evaluated in vitro in seven human cell lines by Western blot, cell counting, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, terminal deoxyribonucleotide transferase-mediated nick-end labeling assays, and fluorescence-activated cell sorting analysis, using two PI3K inhibitors, LY294002 and wortmannin, as well as by transfection with various Akt constructs and through Akt knockdown by small interfering RNA (siRNA). In vivo nude mice bearing human renal cell carcinoma tumor xenografts were treated with LY294002 (75 mg/kg/wk, 4 weeks, i.p.). Tumor growth was measured and tumors were subjected to Western blot and immunohistochemical analysis. Akt was constitutively activated in all cell lines. Constitutive phosphorylation of glycogen synthase kinase-3 (GSK-3) was observed in all cell lines, whereas forkhead transcription factor and mammalian target of rapamycin, although expressed, were not constitutively phosphorylated. Exposure to LY294002 or wortmannin decreased Akt activation and GSK-3 phosphorylation and reduced cell growth by up to 70% through induction of cell apoptosis. These effects were confirmed by transfection experiments with Akt constructs or Akt siRNA. Importantly, LY294002 induced up to 50% tumor regression in mice through tumor cell apoptosis. Tumor neovascularization was significantly increased by LY294002 treatment. Blood chemistries showed no adverse effects of the treatment. Our results suggest an important role of PI3K/Akt inhibitors as a potentially useful treatment for patients with renal cell carcinoma.

AB - Metastatic renal cell carcinoma is resistant to current therapies. The phosphoinositide 3-kinase (PI3K)/Akt signaling cascade induces cell growth, cell transformation, and neovascularization. We evaluated whether targeting this pathway could be of therapeutic value against human renal cell carcinoma. The activation of the PI3K/Akt pathway and its role in renal cell carcinoma progression was evaluated in vitro in seven human cell lines by Western blot, cell counting, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, terminal deoxyribonucleotide transferase-mediated nick-end labeling assays, and fluorescence-activated cell sorting analysis, using two PI3K inhibitors, LY294002 and wortmannin, as well as by transfection with various Akt constructs and through Akt knockdown by small interfering RNA (siRNA). In vivo nude mice bearing human renal cell carcinoma tumor xenografts were treated with LY294002 (75 mg/kg/wk, 4 weeks, i.p.). Tumor growth was measured and tumors were subjected to Western blot and immunohistochemical analysis. Akt was constitutively activated in all cell lines. Constitutive phosphorylation of glycogen synthase kinase-3 (GSK-3) was observed in all cell lines, whereas forkhead transcription factor and mammalian target of rapamycin, although expressed, were not constitutively phosphorylated. Exposure to LY294002 or wortmannin decreased Akt activation and GSK-3 phosphorylation and reduced cell growth by up to 70% through induction of cell apoptosis. These effects were confirmed by transfection experiments with Akt constructs or Akt siRNA. Importantly, LY294002 induced up to 50% tumor regression in mice through tumor cell apoptosis. Tumor neovascularization was significantly increased by LY294002 treatment. Blood chemistries showed no adverse effects of the treatment. Our results suggest an important role of PI3K/Akt inhibitors as a potentially useful treatment for patients with renal cell carcinoma.

KW - 1-Phosphatidylinositol 3-Kinase

KW - Animals

KW - Carcinoma, Renal Cell

KW - Cell Growth Processes

KW - Cell Line, Tumor

KW - Chromones

KW - Enzyme Inhibitors

KW - Humans

KW - Kidney Neoplasms

KW - Male

KW - Mice

KW - Morpholines

KW - Oncogene Protein v-akt

KW - Phosphorylation

KW - RNA, Small Interfering

KW - Signal Transduction

KW - Substrate Specificity

KW - Transfection

KW - Xenograft Model Antitumor Assays

U2 - 10.1158/0008-5472.CAN-05-1469

DO - 10.1158/0008-5472.CAN-05-1469

M3 - Article

VL - 66

SP - 5130

EP - 5142

JO - Cancer Research

JF - Cancer Research

SN - 0008-5472

IS - 10

ER -