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Development of rainbow trout hepatoma cell lines: Effect of pro-IGF-I EA4-peptide on morphological changes and anchorage-independent growth

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Summary

Our laboratory has shown previously that recombinant rainbow trout Ea4 (rtEa4)-peptide of pro-insulin-like growth factor-I (pro-IGF-I) exhibited antitumor activities against cancer cell lines derived from various human cancer tissues (Chen et al., 2002; Kuo and Chen, 2002). To confirm that rtEa4-peptide can exhibit the same spectrum of antitumor activities in fish tumor cells, we had developed permanent single-cell clones (RTH1B1A, RTH1B1D, RTH1B2A, and RTH1B2C) from a rainbow trout liver tumor induced by dibenzo[a,l]pyrene treatment. At 135 passages the doubling time of these single-cell clones in CO2-independent medium at 20°C was 3.9, 3.5, 3.0, and 4.5 d, respectively. Reverse transcription-polymerase chain reaction analysis showed that the expression of liver signature genes (e.g., aldolase B, glucose-6-phosphatase [G-6-Pase], phosphoenolpyruvate carboxykinase [PEPCK], hepatic nuclear factor-1 [HNF-I], IGF-I, IGF-II and growth hormone [GH] receptor-2 genes) and CYP1A1 and CYP1A3 genes was detected in these four single-cell clones. Furthermore, results of in vitro colony formation assay in a soft-agar medium showed different degrees of colony formation activities among them. These results confirmed that the single-cell clones were derived from the rainbow trout liver. Treatment of RTH1B1D with recombinant trout Ea4-peptide resulted in the induction of a dose-dependent morphological change and the suppression of colony formation in a soft-agar medium. In addition, both morphological change and reduction of colony formation were also observed in permanent transfectants of RTH1B1D cells carrying a trout Ea4-peptide gene or its human counterpart, hEb-peptide gene. These results confirm our earlier observations that trout pre-IGF-I Ea4-peptide and hEb possess activities counteracting malignant properties of cancer cells in vitro.

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Authors and Affiliations

  1. Department of Molecular and Cell Biology, University of Connecticut, 91 North Eagleville Road, U-3125, 06269-3125, Storrs, Connecticut

    Maria J. Chen, P. Peter Chiou, Bih-Ying Yang, Hung Chieh Lo & Thomas T. Chen

  2. Department of Environmental and Molecular Toxicology, Oregon State University, 97331, Corvallis, Oregon

    Jerry Hendricks & George Bailey

  3. Department of Marine Biotechnology, Kungnung National University, 210-702, Gangnung, South Korea

    Jin-Ki Son

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  1. Maria J. Chen
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  2. P. Peter Chiou
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  3. Bih-Ying Yang
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  4. Hung Chieh Lo
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  5. Jin-Ki Son
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  6. Jerry Hendricks
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  7. George Bailey
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  8. Thomas T. Chen
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Correspondence to Thomas T. Chen.

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Chen, M.J., Chiou, P.P., Yang, BY. et al. Development of rainbow trout hepatoma cell lines: Effect of pro-IGF-I EA4-peptide on morphological changes and anchorage-independent growth. In Vitro Cell.Dev.Biol.-Animal 40, 118–128 (2004). https://doi.org/10.1290/1543-706X(2004)040<0095:AIVSAF>2.0.CO;2

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  • DOI: https://doi.org/10.1290/1543-706X(2004)040<0095:AIVSAF>2.0.CO;2

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