Geldanamycin decreases Raf-1 and Akt levels and induces apoptosis in neuroblastomas

Sunghoon Kim, Junghee Kang, Wanqin Hu, B. Mark Evers, Dai H. Chung

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

Neuroblastomas are the most common extracranial solid tumors of childhood. These tumors are associated with an overall poor prognosis, particularly for advanced stage disease. The benzoquinone ansamycin antibiotic, geldanamycin (GA), exhibits potent antitumor activity in certain cancer cell lines by destabilizing important signal transduction proteins (e.g., Raf-1 and Akt). The purpose of our study was to determine whether GA can alter the expression of Raf-1 and Akt, which have been shown to be critical for neuronal cell survival, and induce apoptosis of neuroblastoma cells. Human neuroblastoma cells (SH-SY5Y, SK-N-SH and LAN-1) were treated with GA for a variable period of time. Cell viability was assessed with MTT assays. Apoptosis was assessed with DNA fragmentation ELISA, TUNEL-flow cytometric assay, Western blot and caspase activities. We found that GA decreases cell viability and induces apoptosis in the SH-SY5Y human neuroblastoma cell line. These effects were mediated through activation of caspase-9 and -3, mitochondrial release of cytochrome c and subsequent PARP cleavage. GA-induced apoptosis was associated with a reduction in the level and activity of Raf-1 and Akt. The importance of these proteins was further demonstrated by induction of apoptosis in SH-SY5Y cells by a combination of U0126 (MEK 1/2 inhibitor) and LY294002 (an inhibitor of P13K). Similar to SH-SY5Y cells, other human neuroblastoma cells (SK-N-SH and LAN-1) were sensitive to the effects of GA-induced apoptosis. Taken together, our findings suggest that GA may be a novel therapeutic agent, which may be effective in the treatment of neuroblastomas.

Original languageEnglish (US)
Pages (from-to)352-359
Number of pages8
JournalInternational Journal of Cancer
Volume103
Issue number3
DOIs
StatePublished - Jan 20 2003
Externally publishedYes

Fingerprint

Neuroblastoma
Apoptosis
Local Area Networks
Cell Survival
Rifabutin
Cell Line
Neoplasms
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Caspase 9
Mitogen-Activated Protein Kinase Kinases
In Situ Nick-End Labeling
DNA Fragmentation
Caspases
Cytochromes c
geldanamycin
Caspase 3
Signal Transduction
Proteins
Western Blotting
Enzyme-Linked Immunosorbent Assay

Keywords

  • Akt
  • Apoptosis
  • Geldanamycin
  • Neuroblastoma
  • Raf-1

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Geldanamycin decreases Raf-1 and Akt levels and induces apoptosis in neuroblastomas. / Kim, Sunghoon; Kang, Junghee; Hu, Wanqin; Evers, B. Mark; Chung, Dai H.

In: International Journal of Cancer, Vol. 103, No. 3, 20.01.2003, p. 352-359.

Research output: Contribution to journalArticle

Kim, Sunghoon ; Kang, Junghee ; Hu, Wanqin ; Evers, B. Mark ; Chung, Dai H. / Geldanamycin decreases Raf-1 and Akt levels and induces apoptosis in neuroblastomas. In: International Journal of Cancer. 2003 ; Vol. 103, No. 3. pp. 352-359.
@article{5f00db9044a24155b4ba942aa6c0b000,
title = "Geldanamycin decreases Raf-1 and Akt levels and induces apoptosis in neuroblastomas",
abstract = "Neuroblastomas are the most common extracranial solid tumors of childhood. These tumors are associated with an overall poor prognosis, particularly for advanced stage disease. The benzoquinone ansamycin antibiotic, geldanamycin (GA), exhibits potent antitumor activity in certain cancer cell lines by destabilizing important signal transduction proteins (e.g., Raf-1 and Akt). The purpose of our study was to determine whether GA can alter the expression of Raf-1 and Akt, which have been shown to be critical for neuronal cell survival, and induce apoptosis of neuroblastoma cells. Human neuroblastoma cells (SH-SY5Y, SK-N-SH and LAN-1) were treated with GA for a variable period of time. Cell viability was assessed with MTT assays. Apoptosis was assessed with DNA fragmentation ELISA, TUNEL-flow cytometric assay, Western blot and caspase activities. We found that GA decreases cell viability and induces apoptosis in the SH-SY5Y human neuroblastoma cell line. These effects were mediated through activation of caspase-9 and -3, mitochondrial release of cytochrome c and subsequent PARP cleavage. GA-induced apoptosis was associated with a reduction in the level and activity of Raf-1 and Akt. The importance of these proteins was further demonstrated by induction of apoptosis in SH-SY5Y cells by a combination of U0126 (MEK 1/2 inhibitor) and LY294002 (an inhibitor of P13K). Similar to SH-SY5Y cells, other human neuroblastoma cells (SK-N-SH and LAN-1) were sensitive to the effects of GA-induced apoptosis. Taken together, our findings suggest that GA may be a novel therapeutic agent, which may be effective in the treatment of neuroblastomas.",
keywords = "Akt, Apoptosis, Geldanamycin, Neuroblastoma, Raf-1",
author = "Sunghoon Kim and Junghee Kang and Wanqin Hu and Evers, {B. Mark} and Chung, {Dai H.}",
year = "2003",
month = "1",
day = "20",
doi = "10.1002/ijc.10820",
language = "English (US)",
volume = "103",
pages = "352--359",
journal = "International Journal of Cancer",
issn = "0020-7136",
publisher = "Wiley-Liss Inc.",
number = "3",

}

TY - JOUR

T1 - Geldanamycin decreases Raf-1 and Akt levels and induces apoptosis in neuroblastomas

AU - Kim, Sunghoon

AU - Kang, Junghee

AU - Hu, Wanqin

AU - Evers, B. Mark

AU - Chung, Dai H.

PY - 2003/1/20

Y1 - 2003/1/20

N2 - Neuroblastomas are the most common extracranial solid tumors of childhood. These tumors are associated with an overall poor prognosis, particularly for advanced stage disease. The benzoquinone ansamycin antibiotic, geldanamycin (GA), exhibits potent antitumor activity in certain cancer cell lines by destabilizing important signal transduction proteins (e.g., Raf-1 and Akt). The purpose of our study was to determine whether GA can alter the expression of Raf-1 and Akt, which have been shown to be critical for neuronal cell survival, and induce apoptosis of neuroblastoma cells. Human neuroblastoma cells (SH-SY5Y, SK-N-SH and LAN-1) were treated with GA for a variable period of time. Cell viability was assessed with MTT assays. Apoptosis was assessed with DNA fragmentation ELISA, TUNEL-flow cytometric assay, Western blot and caspase activities. We found that GA decreases cell viability and induces apoptosis in the SH-SY5Y human neuroblastoma cell line. These effects were mediated through activation of caspase-9 and -3, mitochondrial release of cytochrome c and subsequent PARP cleavage. GA-induced apoptosis was associated with a reduction in the level and activity of Raf-1 and Akt. The importance of these proteins was further demonstrated by induction of apoptosis in SH-SY5Y cells by a combination of U0126 (MEK 1/2 inhibitor) and LY294002 (an inhibitor of P13K). Similar to SH-SY5Y cells, other human neuroblastoma cells (SK-N-SH and LAN-1) were sensitive to the effects of GA-induced apoptosis. Taken together, our findings suggest that GA may be a novel therapeutic agent, which may be effective in the treatment of neuroblastomas.

AB - Neuroblastomas are the most common extracranial solid tumors of childhood. These tumors are associated with an overall poor prognosis, particularly for advanced stage disease. The benzoquinone ansamycin antibiotic, geldanamycin (GA), exhibits potent antitumor activity in certain cancer cell lines by destabilizing important signal transduction proteins (e.g., Raf-1 and Akt). The purpose of our study was to determine whether GA can alter the expression of Raf-1 and Akt, which have been shown to be critical for neuronal cell survival, and induce apoptosis of neuroblastoma cells. Human neuroblastoma cells (SH-SY5Y, SK-N-SH and LAN-1) were treated with GA for a variable period of time. Cell viability was assessed with MTT assays. Apoptosis was assessed with DNA fragmentation ELISA, TUNEL-flow cytometric assay, Western blot and caspase activities. We found that GA decreases cell viability and induces apoptosis in the SH-SY5Y human neuroblastoma cell line. These effects were mediated through activation of caspase-9 and -3, mitochondrial release of cytochrome c and subsequent PARP cleavage. GA-induced apoptosis was associated with a reduction in the level and activity of Raf-1 and Akt. The importance of these proteins was further demonstrated by induction of apoptosis in SH-SY5Y cells by a combination of U0126 (MEK 1/2 inhibitor) and LY294002 (an inhibitor of P13K). Similar to SH-SY5Y cells, other human neuroblastoma cells (SK-N-SH and LAN-1) were sensitive to the effects of GA-induced apoptosis. Taken together, our findings suggest that GA may be a novel therapeutic agent, which may be effective in the treatment of neuroblastomas.

KW - Akt

KW - Apoptosis

KW - Geldanamycin

KW - Neuroblastoma

KW - Raf-1

UR - http://www.scopus.com/inward/record.url?scp=0037454761&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037454761&partnerID=8YFLogxK

U2 - 10.1002/ijc.10820

DO - 10.1002/ijc.10820

M3 - Article

C2 - 12471618

AN - SCOPUS:0037454761

VL - 103

SP - 352

EP - 359

JO - International Journal of Cancer

JF - International Journal of Cancer

SN - 0020-7136

IS - 3

ER -