Development of rainbow trout hepatoma cell lines: effect of pro-IGF-I Ea4-peptide on morphological changes and anchorage-independent growth.

Abstract

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.

• Authors: Maria J Chen, P Peter Chiou, Bih-Ying Yang, Hung Chieh Lo, Jin-Ki Son, Jerry Hendricks, George Bailey, Thomas T Chen
• Journal: In vitro cellular & developmental biology. Animal (In Vitro Cell Dev Biol Anim) 2004 Mar-Apr Vol. 40 Issue 3-4 Pg. 118-28 ISSN: 1071-2690 [Print] Germany
• PMID: 15311963 (Publication Type: Journal Article, Research Support, U.S. Gov't, Non-P.H.S., Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Benzopyrenes
• Biomarkers, Tumor
• Neoplasm Proteins
• Peptide Fragments
• Protein Precursors
• RNA, Messenger
• Recombinant Proteins
• pro-insulin-like growth factor I
• Insulin-Like Growth Factor I
• dibenzo(a,l)pyrene

Topics

• Animals
• Benzopyrenes (toxicity)
• Biomarkers, Tumor (genetics, metabolism)
• Cell Division (drug effects)
• Cell Line, Tumor
• Gene Expression Regulation, Neoplastic (drug effects)
• Insulin-Like Growth Factor I (therapeutic use)
• Liver Neoplasms, Experimental (drug therapy, metabolism, pathology)
• Neoplasm Proteins (genetics, metabolism)
• Oncorhynchus mykiss
• Peptide Fragments (therapeutic use)
• Protein Precursors (therapeutic use)
• RNA, Messenger (metabolism)
• Recombinant Proteins (therapeutic use)
• Reverse Transcriptase Polymerase Chain Reaction (methods)
• Tumor Stem Cell Assay