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Differential susceptibility of renal carcinoma cell lines to tumor suppression by exogenous Fhit expression
N S Werner1, Z Siprashvili, L Y Fong
1Innere Klinik und Poliklinik (Tumorforschung), Universitätsklinikum Essen, Germany.
Abstract:
Hemizygous deletions of the fragile histidine triad (FHIT) gene at human chromosome band 3p14.2 and down-regulation of its gene product are found in the majority of renal cell carcinomas (RCCs). Functional tumor suppressive activity of Fhit in renal cancer cells previously was observed in RCC cell line RC48, which lacks endogenous Fhit expression. To further investigate the potential role of FHIT as a tumor suppressor gene in RCC, we transfected FHIT cDNA expression constructs into RCC cell lines RCC-1 and SN12C, which show low-level expression of endogenous Fhit and reveal an intact von Hippel-Lindau (VHL) gene. Stable transfectants of both cell lines showed no alterations of cell morphology, proliferation kinetics, or cell cycle parameters in vitro. The FHIT gene transfer rate, however, was significantly lower in RCC-1 cells compared with SN12C cells, suggesting a selection against exogenous Fhit expression. In addition, in nude mouse assays, a significant delay of tumor formation was observed for FHIT-transfected RCC-1 cell lines, with outgrowing tumors demonstrating loss of Fhit expression in the majority of cells. In contrast, tumorigenicity of FHIT-transfected SN12C cell clones was not suppressed, despite stable transgene expression. In conclusion, our results demonstrate a selective tumor suppressive activity of Fhit in RCC cells in vivo and suggest that the susceptibility to suppression is not restricted to cancer cells with complete loss of Fhit expression.
Insights
The fragile histidine triad (FHIT) gene shows selective tumor suppressive activity in renal cell carcinoma (RCC) cells. FHIT gene transfer delayed tumor formation in some RCC cells, indicating its potential role in cancer suppression.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- The fragile histidine triad (FHIT) gene is frequently deleted or downregulated in renal cell carcinomas (RCCs).
- Previous studies suggested a tumor suppressive role for Fhit in renal cancer cells lacking endogenous expression.
Purpose of the Study:
- To investigate the tumor suppressive potential of the FHIT gene in RCC.
- To examine the impact of FHIT reintroduction in RCC cell lines with intact VHL genes.
Main Methods:
- Transfection of FHIT cDNA into RCC cell lines (RCC-1 and SN12C).
- In vitro analysis of cell morphology, proliferation, and cell cycle.
- In vivo tumor formation assays in nude mice.
- Assessment of transgene expression and gene loss in tumors.
Main Results:
- FHIT gene transfer was less efficient in RCC-1 cells, suggesting selection against expression.
- FHIT-transfected RCC-1 cells showed delayed tumor formation in vivo, with subsequent loss of Fhit expression.
- FHIT-transfected SN12C cells did not exhibit suppressed tumorigenicity despite stable transgene expression.
Conclusions:
- Fhit exhibits selective tumor suppressive activity in RCC cells in vivo.
- The susceptibility to FHIT-mediated suppression is not limited to cancer cells with complete FHIT loss.