Development of rainbow trout hepatoma cell lines: effect of pro-IGF-I Ea4-peptide on morphological changes and anchorage-independent growth
| Autor: | Maria J, Chen, P Peter, Chiou, Bih-Ying, Yang, Hung Chieh, Lo, Jin-Ki, Son, Jerry, Hendricks, George, Bailey, Thomas T, Chen |
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| Rok vydání: | 2004 |
| Předmět: |
Reverse Transcriptase Polymerase Chain Reaction
Peptide Fragments Recombinant Proteins Neoplasm Proteins Gene Expression Regulation Neoplastic Liver Neoplasms Experimental Cell Line Tumor Oncorhynchus mykiss Biomarkers Tumor Animals RNA Messenger Benzopyrenes Insulin-Like Growth Factor I Protein Precursors Cell Division Tumor Stem Cell Assay |
| Zdroj: | In vitro cellulardevelopmental biology. Animal. 40(3-4) |
| ISSN: | 1071-2690 |
| Popis: | 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 degrees 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. |
| Databáze: | OpenAIRE |
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