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Identification and expression of human c-Ha-ras and c-sis sequences in NIH3T3 transformants
1Cancer Research Institute, Slovak Academy of Sciences, Bratislava, Czechoslovakia.
Abstract:
High-molecular-weight DNAs from 5 bladder carcinomas were used in transfection of mouse NIH3T3 cells. The manifestation of heterologous oncogene(s) expression in NIH3T3 cells was morphological transformation very often accompanied by changes in growth characteristics of recipient cells. In DNA samples from secondary NIH3T3 transformants human c-Ha-ras and c-sis sequences were identified. In some secondary transformants these sequences were expressed. On the basis of change of the growth characteristics of some secondary transformants we could expect the integration and expression of another human gene(s) for growth factor or growth factor receptor or even activation of mouse genes. We did not manage to identify any Alu sequences in some secondary transformants carrying human c-Ha-ras sequences. On the other hand, it has not been revealed yet that BamHI DNA fragments carrying c-Ha-ras gene contained any Alu sequence. So, the identification of Alu sequences does not have to be the first step in investigation of DNA samples from NIH3T3 transformants.
Insights
Transfecting mouse cells with bladder cancer DNA identified human oncogenes like c-Ha-ras and c-sis, causing cell transformation. Alu sequence presence was inconsistent, suggesting it
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Bladder carcinomas harbor genetic alterations that can drive cancer development.
- Identifying oncogenes and their expression is crucial for understanding cancer progression.
Purpose of the Study:
- To investigate the transforming potential of DNA from bladder carcinomas.
- To identify human oncogenes and their expression in transfected mouse cells.
Main Methods:
- Transfection of mouse NIH3T3 cells with high-molecular-weight DNA from bladder carcinomas.
- Analysis of secondary transformants for human gene sequences (c-Ha-ras, c-sis, Alu) and expression.
- Assessment of morphological and growth characteristic changes in recipient cells.
Main Results:
- NIH3T3 cells transfected with bladder cancer DNA exhibited morphological transformation and altered growth.
- Human c-Ha-ras and c-sis sequences were identified in secondary transformants.
- Expression of human c-Ha-ras and c-sis sequences was observed in some transformants.
- Alu sequences were not consistently detected in transformants carrying human c-Ha-ras.
Conclusions:
- DNA from bladder carcinomas can induce cell transformation in NIH3T3 cells.
- Human c-Ha-ras and c-sis oncogenes are implicated in bladder cancer-induced transformation.
- Alu sequence detection may not be a reliable initial step in analyzing NIH3T3 transformants.