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Dual action of a selective cyclooxygenase-2 inhibitor on vascular endothelial growth factor expression in human hepatocellular carcinoma cells: novel involvement of discoidin domain receptor 2

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Abstract

Purpose

Vascular endothelial growth factor (VEGF) greatly contributes to the progression of hepatocellular carcinoma (HCC). It is reported that a selective cyclooxygenase-2 (COX-2) inhibitor inhibits cellular proliferation and may attenuate VEGF expression in HCC. We propose that different cascades in the VEGF pathway respond to COX-2 inhibition, depending on the cell types.

Methods

The six human HCC cell lines—Hep3B, SNU387, SNU182, SNU423, SNU449, and PLC/PRF5—were cultured under normoxic and hypoxic conditions. Cells were treated with a selective COX-2 inhibitor (NS-398) and discoidin domain receptor 2 (DDR2) siRNA, and microarray analysis was performed.

Results

NS-398 inhibited HCC proliferation and decreased the expression level of VEGF in HCC cells only under normoxia conditions. In hypoxia conditions, VEGF expression level in Hep3B cell was suppressed, while that in SNU387 cell was increased by NS-398 (P < 0.001). The NS-398-induced increase in VEGF expression in SNU387 cell was associated with the up-regulation of the DDR2 gene. NS-398-treated SNU series cells and PLC/PRF5 cells displayed a robust increase in DDR2 mRNA expression. Also, transfection with DDR2 siRNA decreased the VEGF expression level of SNU387, 423, 449 cells under hypoxia conditions (P < 0.05). In vivo chromatin immunoprecipitation assay demonstrated that NS-398 induces the enhancement of HIF-1α binding on VEGF promoter, leading to the increase in VEGF gene expression in hypoxic conditions. There is strong evidence that it is related to the DDR2 gene expression in SNU387 cells.

Conclusion

These findings disclose a novel cell-dependent regulatory mechanism of VEGF involving DDR2 gene in HCC cells.

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Abbreviations

COX-2:

Cyclooxygenase-2

DDR2:

Discoidin domain receptor 2

NS-398:

[N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide], COX-2 inhibitor

VEGF:

Vascular endothelial growth factor

siRNA:

Small interfering RNA

HIF-1α:

Hypoxia-inducible transcription factor 1α

ChIP:

Chromatin immunoprecipitation

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Acknowledgment

This study was supported by a grant from the National Cancer Center, Korea (#0810260, #1110050).

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Authors and Affiliations

  1. Liver and Pancreatobiliary Cancer Research Branch, National Cancer Center, Goyang, South Korea

    Nam Oak Lee, Joong-Won Park & Jung Ahn Lee

  2. Center for Liver Cancer, National Cancer Center, 323 Ilsan-ro, Ilsan dong-gu, Goyang, Gyeonggi, 411-769, South Korea

    Joong-Won Park & Ju Hyun Shim

  3. Center for Clinical Services, National Cancer Center, Goyang, South Korea

    Sun-Young Kong

  4. Functional Genomics Branch, National Cancer Center, Goyang, South Korea

    Kyung Tae Kim & Yeon-Su Lee

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  1. Nam Oak Lee
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Correspondence to Joong-Won Park.

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Lee, N.O., Park, JW., Lee, J.A. et al. Dual action of a selective cyclooxygenase-2 inhibitor on vascular endothelial growth factor expression in human hepatocellular carcinoma cells: novel involvement of discoidin domain receptor 2. J Cancer Res Clin Oncol 138, 73–84 (2012). https://doi.org/10.1007/s00432-011-1075-0

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  • DOI: https://doi.org/10.1007/s00432-011-1075-0

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