Activation of hypoxia-inducible factor-1α is necessary for lysophosphatidic acid-induced vascular endothelial growth factor expression

Jangsoon Lee, Soon Young Park, Eun Kyung Lee, Chang Gyo Park, Hyun Cheol Chung, Sun Young Rha, Yong Kee Kim, Gyu Un Bae, Bum Kyeong Kim, Jeung Whan Han, Hoi Young Lee

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

Purpose: Lysophosphatidic acid (LPA) plays an important role in mediating cell proliferation, survival, and tumor invasion and angiogenesis. This bioactive phospholipid at the concentration in ascitic fluid stimulates the growth of malignant ovarian tumors by increasing the expression of vascular endothelial growth factor (VEGF). In the present study, we investigated whether LPA activates hypoxia inducible factor-1 (HIF-1), a key transcriptional complex in tumor progression and metastasis, thereby increasing the expression of VEGF. Experimental Design: Immunoblotting, reverse transcription-PCR, ELISA, immunofluorescence, and chromatin immunoprecipitation assay were used to examine the expression of VEGF and HIF-1 α in various cancer cells. Specific HIF-1 α small interfering RNA was transfected to various cancer cells to determine the role of HIF-1 α in LPA-induced VEGF expression. Results: LPA induced expressions of VEGF and HIF-1 α in OVCAR-3, CAOV-3, PC-3, and SK-Hep1 cells but not in SKOV-3 and Hep-3B cells. In OVCAR-3 and PC-3 cells, the phosphoinositide 3-kinase/Akt/mammalian target of rapamycin/p70S6K and p42/p44 mitogen-activated protein kinase pathways were required for LPA-induced HIF-1 α and VEGF expressions, whereas only the phosphoinositide 3-kinase/mammalian target of rapamycin/p70S6K pathway was important in SK-Hep1 cells. Immunofluorescence microscopy assay showed translocation of HIF-1 α to nucleus by LPA, and chromatin immunoprecipitation assay revealed the binding of HIF-1 α to the promoter of VEGF by LPA. Importantly, we found that small interfering RNA-induced reduction of HIF-1 α expression significantly attenuated VEGF expression by LPA. Conclusions: Our results show for the first time that LPA induces VEGF via HIF-1 α activation and reveal a critical role of HIF-1 α in LPA-induced cancer cell proliferation and angiogenesis.

Original languageEnglish
Pages (from-to)6351-6358
Number of pages8
JournalClinical Cancer Research
Volume12
Issue number21
DOIs
Publication statusPublished - 2006 Nov 1

Fingerprint

Hypoxia-Inducible Factor 1
Vascular Endothelial Growth Factor A
70-kDa Ribosomal Protein S6 Kinases
Neoplasms
1-Phosphatidylinositol 4-Kinase
Chromatin Immunoprecipitation
Sirolimus
Small Interfering RNA
lysophosphatidic acid
Cell Proliferation
Ascitic Fluid
Mitogen-Activated Protein Kinase 1
Fluorescence Microscopy
Immunoblotting
Reverse Transcription
Fluorescent Antibody Technique
Phospholipids
Cell Survival
Research Design
Enzyme-Linked Immunosorbent Assay

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

Lee, Jangsoon ; Park, Soon Young ; Lee, Eun Kyung ; Park, Chang Gyo ; Chung, Hyun Cheol ; Rha, Sun Young ; Kim, Yong Kee ; Bae, Gyu Un ; Kim, Bum Kyeong ; Han, Jeung Whan ; Lee, Hoi Young. / Activation of hypoxia-inducible factor-1α is necessary for lysophosphatidic acid-induced vascular endothelial growth factor expression. In: Clinical Cancer Research. 2006 ; Vol. 12, No. 21. pp. 6351-6358.
@article{0892482547e54171bb3ecc8bd1c94794,
title = "Activation of hypoxia-inducible factor-1α is necessary for lysophosphatidic acid-induced vascular endothelial growth factor expression",
abstract = "Purpose: Lysophosphatidic acid (LPA) plays an important role in mediating cell proliferation, survival, and tumor invasion and angiogenesis. This bioactive phospholipid at the concentration in ascitic fluid stimulates the growth of malignant ovarian tumors by increasing the expression of vascular endothelial growth factor (VEGF). In the present study, we investigated whether LPA activates hypoxia inducible factor-1 (HIF-1), a key transcriptional complex in tumor progression and metastasis, thereby increasing the expression of VEGF. Experimental Design: Immunoblotting, reverse transcription-PCR, ELISA, immunofluorescence, and chromatin immunoprecipitation assay were used to examine the expression of VEGF and HIF-1 α in various cancer cells. Specific HIF-1 α small interfering RNA was transfected to various cancer cells to determine the role of HIF-1 α in LPA-induced VEGF expression. Results: LPA induced expressions of VEGF and HIF-1 α in OVCAR-3, CAOV-3, PC-3, and SK-Hep1 cells but not in SKOV-3 and Hep-3B cells. In OVCAR-3 and PC-3 cells, the phosphoinositide 3-kinase/Akt/mammalian target of rapamycin/p70S6K and p42/p44 mitogen-activated protein kinase pathways were required for LPA-induced HIF-1 α and VEGF expressions, whereas only the phosphoinositide 3-kinase/mammalian target of rapamycin/p70S6K pathway was important in SK-Hep1 cells. Immunofluorescence microscopy assay showed translocation of HIF-1 α to nucleus by LPA, and chromatin immunoprecipitation assay revealed the binding of HIF-1 α to the promoter of VEGF by LPA. Importantly, we found that small interfering RNA-induced reduction of HIF-1 α expression significantly attenuated VEGF expression by LPA. Conclusions: Our results show for the first time that LPA induces VEGF via HIF-1 α activation and reveal a critical role of HIF-1 α in LPA-induced cancer cell proliferation and angiogenesis.",
author = "Jangsoon Lee and Park, {Soon Young} and Lee, {Eun Kyung} and Park, {Chang Gyo} and Chung, {Hyun Cheol} and Rha, {Sun Young} and Kim, {Yong Kee} and Bae, {Gyu Un} and Kim, {Bum Kyeong} and Han, {Jeung Whan} and Lee, {Hoi Young}",
year = "2006",
month = "11",
day = "1",
doi = "10.1158/1078-0432.CCR-06-1252",
language = "English",
volume = "12",
pages = "6351--6358",
journal = "Clinical Cancer Research",
issn = "1078-0432",
publisher = "American Association for Cancer Research Inc.",
number = "21",

}

Activation of hypoxia-inducible factor-1α is necessary for lysophosphatidic acid-induced vascular endothelial growth factor expression. / Lee, Jangsoon; Park, Soon Young; Lee, Eun Kyung; Park, Chang Gyo; Chung, Hyun Cheol; Rha, Sun Young; Kim, Yong Kee; Bae, Gyu Un; Kim, Bum Kyeong; Han, Jeung Whan; Lee, Hoi Young.

In: Clinical Cancer Research, Vol. 12, No. 21, 01.11.2006, p. 6351-6358.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Activation of hypoxia-inducible factor-1α is necessary for lysophosphatidic acid-induced vascular endothelial growth factor expression

AU - Lee, Jangsoon

AU - Park, Soon Young

AU - Lee, Eun Kyung

AU - Park, Chang Gyo

AU - Chung, Hyun Cheol

AU - Rha, Sun Young

AU - Kim, Yong Kee

AU - Bae, Gyu Un

AU - Kim, Bum Kyeong

AU - Han, Jeung Whan

AU - Lee, Hoi Young

PY - 2006/11/1

Y1 - 2006/11/1

N2 - Purpose: Lysophosphatidic acid (LPA) plays an important role in mediating cell proliferation, survival, and tumor invasion and angiogenesis. This bioactive phospholipid at the concentration in ascitic fluid stimulates the growth of malignant ovarian tumors by increasing the expression of vascular endothelial growth factor (VEGF). In the present study, we investigated whether LPA activates hypoxia inducible factor-1 (HIF-1), a key transcriptional complex in tumor progression and metastasis, thereby increasing the expression of VEGF. Experimental Design: Immunoblotting, reverse transcription-PCR, ELISA, immunofluorescence, and chromatin immunoprecipitation assay were used to examine the expression of VEGF and HIF-1 α in various cancer cells. Specific HIF-1 α small interfering RNA was transfected to various cancer cells to determine the role of HIF-1 α in LPA-induced VEGF expression. Results: LPA induced expressions of VEGF and HIF-1 α in OVCAR-3, CAOV-3, PC-3, and SK-Hep1 cells but not in SKOV-3 and Hep-3B cells. In OVCAR-3 and PC-3 cells, the phosphoinositide 3-kinase/Akt/mammalian target of rapamycin/p70S6K and p42/p44 mitogen-activated protein kinase pathways were required for LPA-induced HIF-1 α and VEGF expressions, whereas only the phosphoinositide 3-kinase/mammalian target of rapamycin/p70S6K pathway was important in SK-Hep1 cells. Immunofluorescence microscopy assay showed translocation of HIF-1 α to nucleus by LPA, and chromatin immunoprecipitation assay revealed the binding of HIF-1 α to the promoter of VEGF by LPA. Importantly, we found that small interfering RNA-induced reduction of HIF-1 α expression significantly attenuated VEGF expression by LPA. Conclusions: Our results show for the first time that LPA induces VEGF via HIF-1 α activation and reveal a critical role of HIF-1 α in LPA-induced cancer cell proliferation and angiogenesis.

AB - Purpose: Lysophosphatidic acid (LPA) plays an important role in mediating cell proliferation, survival, and tumor invasion and angiogenesis. This bioactive phospholipid at the concentration in ascitic fluid stimulates the growth of malignant ovarian tumors by increasing the expression of vascular endothelial growth factor (VEGF). In the present study, we investigated whether LPA activates hypoxia inducible factor-1 (HIF-1), a key transcriptional complex in tumor progression and metastasis, thereby increasing the expression of VEGF. Experimental Design: Immunoblotting, reverse transcription-PCR, ELISA, immunofluorescence, and chromatin immunoprecipitation assay were used to examine the expression of VEGF and HIF-1 α in various cancer cells. Specific HIF-1 α small interfering RNA was transfected to various cancer cells to determine the role of HIF-1 α in LPA-induced VEGF expression. Results: LPA induced expressions of VEGF and HIF-1 α in OVCAR-3, CAOV-3, PC-3, and SK-Hep1 cells but not in SKOV-3 and Hep-3B cells. In OVCAR-3 and PC-3 cells, the phosphoinositide 3-kinase/Akt/mammalian target of rapamycin/p70S6K and p42/p44 mitogen-activated protein kinase pathways were required for LPA-induced HIF-1 α and VEGF expressions, whereas only the phosphoinositide 3-kinase/mammalian target of rapamycin/p70S6K pathway was important in SK-Hep1 cells. Immunofluorescence microscopy assay showed translocation of HIF-1 α to nucleus by LPA, and chromatin immunoprecipitation assay revealed the binding of HIF-1 α to the promoter of VEGF by LPA. Importantly, we found that small interfering RNA-induced reduction of HIF-1 α expression significantly attenuated VEGF expression by LPA. Conclusions: Our results show for the first time that LPA induces VEGF via HIF-1 α activation and reveal a critical role of HIF-1 α in LPA-induced cancer cell proliferation and angiogenesis.

UR - http://www.scopus.com/inward/record.url?scp=33751297349&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33751297349&partnerID=8YFLogxK

U2 - 10.1158/1078-0432.CCR-06-1252

DO - 10.1158/1078-0432.CCR-06-1252

M3 - Article

C2 - 17085645

AN - SCOPUS:33751297349

VL - 12

SP - 6351

EP - 6358

JO - Clinical Cancer Research

JF - Clinical Cancer Research

SN - 1078-0432

IS - 21

ER -