Differential display screening was used to reveal differential gene expression between the tumorigenic breast cancer cell line CAL51 and nontumorigenic microcell hybrids obtained after transfer of human chromosome 17 into CAL51. The human profilin 1 (PFN1) gene was found overexpressed in the microcell hybrid clones compared with the parental line, which displayed a low profilin 1 level. A comparison between several different tumorigenic breast cancer cell lines with nontumorigenic lines showed consistently lower profilin 1 levels in the tumor cells. Transfection of PFN1 cDNA into CAL51 cells raised the profilin 1 level, had a prominent effect on cell growth, cytoskeletal organization and spreading, and suppressed tumorigenicity of the stable, PFN1-overexpressing cell clones in nude mice. Immunohistochemical analysis revealed intermediate and low levels of profilin 1 in different human breast cancers. These results suggest profilin 1 as a suppressor of the tumorigenic phenotype of breast cancer cells.
Infection of Chinese hamster cells with SV 40 DNA gives rise to mutants resistant both to S-axaguanine (AG) and aminopterin (AP). This mutagenic effect can be raised when facilitating DNA uptake of cells by a helper agent. The extent of muyagenic action depends further on the concentration of DNA applied to the cells, with 2 micrograms/ml being more effective than 10 micrograms/ml, as well as on the period of incubation of infected cells before onset of mutant selection (mutation expression time). Using the AG resistance marker the mutation frequency can be increased more than 8-fold compared with the spontaneous mutation frequency. Reconstituted SV 40 minichromosomes show a mutagenic action which is similar to the DNA-mediated mutagensis whereas non-viral DNA from mammalian cells fails to induce mutations significantly. A major part of isolated clones of SV 40-induced mutants tested so far does express SV 40 T-antigen, suggesting the persistence of SV 40 genetic material in these clones. The possible existence of relations between mutagenic and transforming capacities of SV 40 is discussed.
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