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. 2010 May 11;102(10):1483-90.
doi: 10.1038/sj.bjc.6605669. Epub 2010 Apr 20.

Insulin-like growth factor-binding protein 7 alters the sensitivity to interferon-based anticancer therapy in hepatocellular carcinoma cells

Y Tomimaru  1 , H Eguchi, H Wada, T Noda, M Murakami, S Kobayashi, S Marubashi, Y Takeda, M Tanemura, K Umeshita, Y Doki, M Mori, H Nagano
Affiliations

Insulin-like growth factor-binding protein 7 alters the sensitivity to interferon-based anticancer therapy in hepatocellular carcinoma cells

Y Tomimaru et al. Br J Cancer. .

Abstract

Background: A striking efficiency of interferon (IFN)-based anticancer therapy for advanced hepatocellular carcinoma (HCC) has been reported. Because its clinical efficiency greatly depends on each patient's local response, prediction of local response is crucial.

Methods: Continuous exposure of IFN-alpha to parental PLC/PRF/5 cells (PLC-P) and a limiting dilution method resulted in the establishment of IFN-resistant cell clones (PLC-Rs). Microarray analyses of PLC-P and PLC-Rs identified insulin-like growth factor-binding protein 7 (IGFBP7) as one of the most significantly downregulated genes in PLC-Rs. Changes in anticancer effects of IFN-alpha were examined in HCC cells after genetic manipulation of IGFBP7 expression. The correlation between immunohistochemically determined IGFBP7 expression and the response to IFN-alpha/5-fluorouracil (5-FU) therapy was investigated in surgically resected HCC specimens.

Results: PLC-R cells showed a remarkable downregulation of IGFBP7 and resistance to IFN-alpha, compared with PLC-P. Parental PLC/PRF/5 cells transfected with short hairpin RNA against IGFBP7 showed a significant resistance to IFN-alpha relative to control cells (IC(50) fold increase=14.38 times). Insulin-like growth factor-binding protein 7 transfection into PLC-R restored sensitivity to IFN-alpha. In resected specimens, IGFBP7 expression significantly correlated with the response to IFN-alpha/5-FU therapy.

Conclusion: IGFBP7 could be a useful predictor of the response to IFN-based therapy in advanced HCC.

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Figures

Figure 1
Figure 1
Characteristics of IFN-resistant PLC/PRF/5 cell clones (PLC-Rs). (A) MTT assay showed that the antitumour effect of interferon-α (IFN-α) in PLC-Rs was significantly lower than that in parental PLC/PRF/5 cells (PLC-P). Data are mean±s.d. *P<0.05 compared with PLC-P. (B) Western blot analysis revealed that the expression levels of type I IFN receptor type 2 (IFNAR2), signal transducer and activator of transcription factor 1 (STAT1), STAT2, phosphorylated STAT1 (pSTAT1), and pSTAT2 were similar in PLC-P and PLC-Rs. (C) Schematic of the results of the performed microarray analysis and identified 107 genes. The 107 genes were up- or downregulated by more than 1.5-fold and were commonly identified in the three types of cells. (D) Quantitative reverse transcriptase-PCR and western blot analysis confirmed the significant suppression of insulin-like growth factor-binding protein 7 (IGFBP7) expression in PLC-Rs compared with PLC-P. Data are mean±s.d. *P<0.05.
Figure 2
Figure 2
Characteristics of parental PLC/PRF/5 cell (PLC-P)/short hairpin RNA (shRNA) (no. 1 and no. 2). (A) Insulin-like growth factor-binding protein 7 (IGFBP7) was confirmed to be significantly suppressed in PLC-P/shRNA compared with PLC-P/shRNA-negative control (NC) in quantitative reverse transcriptase-PCR and western blot analysis. (B) MTT assay revealed that PLC-P/shRNA was significantly more resistant to interferon-α (IFN-α) than was PLC-P/shRNA-NC. (C) The percentage of early apoptotic PLC-P/shRNA cells assessed by annexin V assay was significantly lower than that of PLC-P/shRNA-NC. (D) The activity of caspase-3, caspase-8, and caspase-9 induced by IFN-α in PLC-P/shRNA was significantly lower than that in PLC-P/shRNA-NC. Data are mean±s.d. *P<0.05.
Figure 3
Figure 3
Characteristics of IFN-resistant PLC/PRF/5 cell clones (PLC-R1) transfected with Insulin-like growth factor-binding protein 7 (IGFBP7) expression plasmid. (A) Quantitative reverse transcriptase-PCR and western blot analysis showed that the IGFBP7 expression level in PLC-R1/IGFBP7 was significantly higher than that in PLC-R1/IGFBP7-negative control (NC). (B) MTT assay showed that PLC-R1/IGFBP7 were significantly more sensitive to IFN-α than was PLC-R1/IGFBP7-NC. Data are mean±s.d. *P<0.05.
Figure 4
Figure 4
Immunohistochemistry for Insulin-like growth factor-binding protein 7 (IGFBP7) in representative hepatocellular carcinoma cases (A) A representative IGFBP7-positive case. The IGFBP7 expression was shown in the cytoplasm of normal liver cells and in the majority of tumour cells. (B) A representative IGFBP7-negative case. The IGFBP7 expression was not identified in tumour cells. Upper panel, low-power field (Bar=200 μm); lower panel, high-power field (Bar=50 μm); T, tumour lesion (arrowheads); N, non-tumour lesion.
Figure 5
Figure 5
Postoperative overall survival curves showed a significantly better survival rate for Insulin-like growth factor-binding protein 7 (IGFBP7)-positive patients than for IGFBP7-negative patients (*P<0.05).
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References

    1. Asahara T, Itamoto T, Katayama K, Nakahara H, Hino H, Yano M, Ono E, Dohi K, Nakanishi T, Kitamoto M, Azuma K, Itoh K, Shimamoto F (1999) Hepatic resection with tumor thrombectomy for hepatocellular carcinoma with tumor thrombi in the major vasculatures. Hepatogastroenterology 46: 1862–1869 - PubMed
    1. Burger AM, Zhang X, Li H, Ostrowski JL, Beatty B, Venanzoni M, Papas T, Seth A (1998) Down-regulation of T1A12/mac25, a novel insulin-like growth factor binding protein related gene, is associated with disease progression in breast carcinomas. Oncogene 16: 2459–2467 - PubMed
    1. Chung YH, Song IH, Song BC, Lee GC, Koh MS, Yoon HK, Lee YS, Sung KB, Suh DJ (2000) Combined therapy consisting of intraarterial cisplatin infusion and systemic interferon-alpha for hepatocellular carcinoma patients with major portal vein thrombosis or distant metastasis. Cancer 88: 1986–1991 - PubMed
    1. Damdinsuren B, Nagano H, Monden M (2007a) Combined intra-arterial 5-fluorouracil and subcutaneous interferon-α therapy for highly advanced hepatocellular carcinoma. Hepatol Res 37(Suppl 2): S238–S250 - PubMed
    1. Damdinsuren B, Nagano H, Wada H, Noda T, Natsag J, Marubashi S, Miyamoto A, Takeda Y, Umeshita K, Doki Y, Dono K, Monden M (2007b) Interferon α receptors are important for antiproliferative effect of interferon-α against human hepatocellular carcinoma cells. Hepatol Res 37: 77–83 - PubMed

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