NIH 3T3 Cell Transformation by DNAs from Lymphoma Cells and from Epstein-Barr Virus-Immortalized Human Lymphocytes
DNA extracted from P3HRI cells morphologically transforms NIH 3T3 mouse fibroblast cells. We have constructed a gene library of the transformed cell DNA by utilizing lambda-EMBL 4 phage as a cloning vector and isolating sequences which contained human Alu sequences. This DNA was capable of NIH 3T3 cell transformation but did not hybridize to cloned B-lym DNA. Fresh lymphocytes from cord blood or tonsil were stimulated by LPS or immortalized by EBV infection. Immortalized cells were further cloned in 0.45% agarose. The efficiency of colony formation was found to be 0.2–2.0%. DNA from fresh lymphocytes, lymphocytes stimulated with LPS, lymphocytes immortalized by EBV infection, and from cloned cells were tested for NIH 3T3 cell transformation. Same of the fresh lymphocytes already showed a capability of transforming NIH 3T3 cells; on the other hand, same of the EBV-immortalized cloned lymphocytes showed transformation ability. The transformed cells were tumor inducible in nude mice. These results may indicate that a certain population of lymphocytes is already potentially oncogenic by means of either activated oncogene or genomic rearrangement.
KeywordsLymphoma Agarose Prep
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