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Stable oncogenic transformation induced by microcell-mediated gene transfer
Science in China. Series B, Chemistry, Life Sciences & Earth Sciences
|April 1, 1995
Summary
Microcell-mediated gene transfer effectively transfers dominant oncogenes, retaining large, unrearranged DNA segments. This technique aids in studying oncogene function and gene mapping in cell transformation.
Area of Science:
- Molecular Biology
- Genetics
- Cancer Research
Background:
- DNA-mediated transfection has limitations in transferring large, unrearranged DNA segments, hindering oncogene studies.
- Gene rearrangement and amplification during transfection complicate the analysis of oncogenes.
Purpose of the Study:
- To evaluate microcell-mediated gene transfer for analyzing dominant oncogenes.
- To optimize microcell preparation and transfer for studying the met oncogene.
Main Methods:
- Utilized MNNG-HOS cells containing the met oncogene on human chromosome 7.
- Infected cells with retroviruses carrying drug resistance markers.
- Optimized microcell preparation and transfer for creating stable hybrids.
Main Results:
- Obtained drug-resistant hybrids with single human chromosomes and transformed cell foci containing the met oncogene.
- Hybridization analysis showed varying amounts of unrearranged human DNA associated with the oncogene.
- Microcell transformants retained more distal markers compared to DNA- or chromosome-mediated transfers.
Conclusions:
- Microcell-mediated gene transfer is a valuable technique for transferring and analyzing dominant oncogenes.
- This method facilitates gene mapping and the study of oncogene function and expression.
- It overcomes limitations of DNA-mediated transfection regarding DNA integrity and size.