Cyclooxygenase-2 inhibitor and interferon-β synergistically induce apoptosis in human hepatoma cells in vitro and in vivo

Nobuhiro Nakamoto, Hajime Higuchi, Hideaki Kanamori, Satoshi Kurita, Shinichiro Tada, Hiromasa Takaishi, Kyoko Toda, Takaya Yamada, Naoki Kumagai, Hidetsugu Saito, Toshifumi Hibi

Research output: Contribution to journalArticle

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Abstract

Recent clinical trials have shown that interferon (IFN) is effective for chemoprevention against hepatocellular carcinoma (HCC). However, it remains controversial as to whether IFN exerts direct cytotoxicity against HCC. Cyclooxygenase (COX)-2 also plays a role in hepatocarcinogenesis and may mediate resistance to apoptosis in HCC. Therefore, we aimed to elucidate the combined effect of COX-2 inhibitor, NS-398, and IFN on in vitro growth suppression of HCC using 3 hepatoma cell lines (HepG2, PLC/PRF/5, and Huh7) and in vivo nude mouse xenotransplantation model using Huh7 cells. Only minimal growth inhibition was observed after treatment with IFN-β alone in the 3 hepatoma cell lines. In contrast, treatment with NS-398 and IFN-β synergistically inhibited cell proliferation in dose- and time-dependent manner. Apoptosis was identified by 4′,6-diamidino-2-phenylindole dihydrochloride and fluorescent staining. IFN-β up-regulated the expression of TRAIL, while NS-398 increased the expression of TRAIL receptors (especially of death receptor 5). Subsequently, activation of caspase-8 and caspase-3 was observed following the treatment with NS-398 and IFN-β. Blockade of TRAIL with a specific antibody attenuated this apoptosis. Furthermore, we found that IFN-β up-regulated COX-2 expression in Huh7 cells, and NS-398 might suppress the up-regulated COX-2 activity downstream of IFN signaling. In vivo experiment showed the combined regimen with NS-398 and IFN-β reduced the growth of xenotransplated HCCs in nude mice. In conclusion, NS-398 is sufficient to overcome IFN resistance in hepatoma cells through the TRAIL/TRAIL receptor pathway, therefore, the combination would appear to be a new therapeutic regimen for HCC.

Original languageEnglish
Pages (from-to)625-635
Number of pages11
JournalInternational Journal of Oncology
Volume29
Issue number3
Publication statusPublished - 2006 Sep

Fingerprint

Cyclooxygenase 2 Inhibitors
Interferons
Hepatocellular Carcinoma
Apoptosis
TNF-Related Apoptosis-Inducing Ligand Receptors
Cyclooxygenase 2
Nude Mice
In Vitro Techniques
Growth
Cell Line
Heterologous Transplantation
Caspase 8
Chemoprevention
Therapeutics
N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide
Caspase 3
Cell Proliferation
Clinical Trials
Staining and Labeling

Keywords

  • Death receptor
  • Huh7
  • Nude mice
  • Prostaglandin E2
  • TRAIL

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Cyclooxygenase-2 inhibitor and interferon-β synergistically induce apoptosis in human hepatoma cells in vitro and in vivo. / Nakamoto, Nobuhiro; Higuchi, Hajime; Kanamori, Hideaki; Kurita, Satoshi; Tada, Shinichiro; Takaishi, Hiromasa; Toda, Kyoko; Yamada, Takaya; Kumagai, Naoki; Saito, Hidetsugu; Hibi, Toshifumi.

In: International Journal of Oncology, Vol. 29, No. 3, 09.2006, p. 625-635.

Research output: Contribution to journalArticle

Nakamoto, N, Higuchi, H, Kanamori, H, Kurita, S, Tada, S, Takaishi, H, Toda, K, Yamada, T, Kumagai, N, Saito, H & Hibi, T 2006, 'Cyclooxygenase-2 inhibitor and interferon-β synergistically induce apoptosis in human hepatoma cells in vitro and in vivo', International Journal of Oncology, vol. 29, no. 3, pp. 625-635.
Nakamoto, Nobuhiro ; Higuchi, Hajime ; Kanamori, Hideaki ; Kurita, Satoshi ; Tada, Shinichiro ; Takaishi, Hiromasa ; Toda, Kyoko ; Yamada, Takaya ; Kumagai, Naoki ; Saito, Hidetsugu ; Hibi, Toshifumi. / Cyclooxygenase-2 inhibitor and interferon-β synergistically induce apoptosis in human hepatoma cells in vitro and in vivo. In: International Journal of Oncology. 2006 ; Vol. 29, No. 3. pp. 625-635.
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abstract = "Recent clinical trials have shown that interferon (IFN) is effective for chemoprevention against hepatocellular carcinoma (HCC). However, it remains controversial as to whether IFN exerts direct cytotoxicity against HCC. Cyclooxygenase (COX)-2 also plays a role in hepatocarcinogenesis and may mediate resistance to apoptosis in HCC. Therefore, we aimed to elucidate the combined effect of COX-2 inhibitor, NS-398, and IFN on in vitro growth suppression of HCC using 3 hepatoma cell lines (HepG2, PLC/PRF/5, and Huh7) and in vivo nude mouse xenotransplantation model using Huh7 cells. Only minimal growth inhibition was observed after treatment with IFN-β alone in the 3 hepatoma cell lines. In contrast, treatment with NS-398 and IFN-β synergistically inhibited cell proliferation in dose- and time-dependent manner. Apoptosis was identified by 4′,6-diamidino-2-phenylindole dihydrochloride and fluorescent staining. IFN-β up-regulated the expression of TRAIL, while NS-398 increased the expression of TRAIL receptors (especially of death receptor 5). Subsequently, activation of caspase-8 and caspase-3 was observed following the treatment with NS-398 and IFN-β. Blockade of TRAIL with a specific antibody attenuated this apoptosis. Furthermore, we found that IFN-β up-regulated COX-2 expression in Huh7 cells, and NS-398 might suppress the up-regulated COX-2 activity downstream of IFN signaling. In vivo experiment showed the combined regimen with NS-398 and IFN-β reduced the growth of xenotransplated HCCs in nude mice. In conclusion, NS-398 is sufficient to overcome IFN resistance in hepatoma cells through the TRAIL/TRAIL receptor pathway, therefore, the combination would appear to be a new therapeutic regimen for HCC.",
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