mercaptopurine and Cell-Transformation--Viral

The present paper deals with effects of 12-O-tetradecanoylphorbol-13-acetate (TPA) on the frequency of induced mutations to 6-mercaptopurine (6MP) and ouabain resistance in Chinese hamster and mouse cells. UV light, bovine adenovirus 3(BAV-3) and 5-bromodeoxyuridine (BrdU) were used as mutagens. TPA was shown to raise the frequency of gene mutations induced by UV light and BAV-3 but it did not enhance the mutagenic effect of BrdU. We also examined the ability of BAV-3 and BrdU to induce tumours in mice. BrdU was shown to have no carcinogenic effect. The results suggest that TPA enhances the mutagenic effect only for carcinogenic mutagens.

Topics: Adenoviridae; Animals; Bromodeoxyuridine; Cell Line; Cell Transformation, Viral; Cells, Cultured; Cricetinae; Cricetulus; Mercaptopurine; Mice; Mutagens; Mutation; Tetradecanoylphorbol Acetate; Ultraviolet Rays

The mutagenic and transforming activity of SV40 DNA fragment, corresponding to its oncogene (the gene for large T antigen) was studied in Chinese hamster cells. After expression time of 3 to 4 days, the oncogene induced mutations of resistance to 6-mercaptopurine (6MP), while the DNA encoding the SV40 late genes, as well as DNA of Chinese hamster cells, were devoid of mutagenic activity. The value of induction ranged from 10(-4) to 10(-5). After the same expression time, the oncogene induced a typical character of oncogenic transformation - independence of serum growth factors (ser+). The value of induction of ser+ variants was somewhat higher than for resistance mutations. The study of 12 clones induced by the oncogene has shown the ser+ character to be hereditary, the expression of viral oncogene being not necessary for its maintenance. The data obtained support the hypothesis in favour of the participation of mutations of cellular genes in viral carcinogenesis.

Topics: Animals; Antigens, Viral, Tumor; Cell Transformation, Viral; Cells, Cultured; Cricetinae; Cricetulus; DNA, Viral; Drug Resistance; Mercaptopurine; Mutation; Oncogenes; Simian virus 40; Transfection

The mutagenic and carcinogenic effect of two EcoRI-fragments of bovine adenovirus type 3 (BAV-3) DNA inserted into pBR325 has been studied. The C fragment (located between 3,6 and 19,7 map units) contains the viral oncogene, the C fragment (between 44,3 and 63,7 map units) displays no transforming activity. It has been established that oncogene BAV-3 statistically true increases the yield of mutants resistant to 6-mercaptopurine (6MP) in Chinese hamster cells. The C fragment, pBR325 without viral sequences and DNA fragments of different molecular weights from normal Syrian hamster cells have no mutagenic effect. The control over tumor formation in syngenic mice after injection of C3H10T 1/2 and D. C fragments and pBR325 treatment exposed a parallelism between the mutagenic and transforming effect. The study of the combined effect of viral DNA fragments and the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) which increases the transforming activity of different carcinogens, shows that the promoter increases the frequency of mutants after viral oncogene treatment and does not induce mutagenic activity of those types of DNA which are unable to transform the cells.

Topics: Adenoviridae; Animals; Cell Transformation, Neoplastic; Cell Transformation, Viral; Cells, Cultured; Cricetinae; Cricetulus; DNA, Viral; Dose-Response Relationship, Drug; Drug Resistance; Mercaptopurine; Mice; Mice, Inbred C3H; Mutation; Oncogenes; Tetradecanoylphorbol Acetate; Time Factors; Transfection

The genetic events controlling the ability of transformed cells to grow in a medium with a low serum content (ser+) were studied. A hypodiploid clone of Chinese hamster cells with normal serum requirements (49a5ser-) was used as starting material. The results of the fluctuation tests have shown that serum-independence is a random spontaneous event. Its rate of occurrence is 1-2 . 10(-5). The concomitant study of a gene mutation (resistance to 6-mercaptopurine) revealed similar characteristics with respect to the distribution of the number of mutants in replicate cultures and the mutation rate. N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and the oncogenic SV40 virus significantly increased the frequency of ser+ colonies. In the majority of clones isolated in a medium with 1% serum (11 spontaneous and 7 induced by MNNG), the ser+ character proved to be stable after different periods of cultivation without selective pressure. The degree of serum-independence varied in different clones. The results suggest that the ability to grow in a medium with a low serum content originates, in most cases, from a mutation event.

Topics: Animals; Cell Transformation, Neoplastic; Cell Transformation, Viral; Cells, Cultured; Clone Cells; Cricetinae; Cricetulus; Culture Media; Diploidy; Drug Resistance; Genetic Techniques; Mercaptopurine; Methylnitronitrosoguanidine; Mutation; Simian virus 40

The mutagenic activity of the tsA239 mutant of SV40 which synthetizes a defective T antigen at 40 degrees C was investigated in Chinese hamster cells under permissive and nonpermissive temperature. At 33 degrees C the virus increased the yield of 6-mercaptopurine-resistant colonies after 2 days expression time by a factor of 1.6-4 as compared with the control and raised the frequency of aberrant metaphases after the same time by a factor of 1.9-3.4. In the same experiments, with the same initially infected population of Chinese hamster cells, at 40 degrees C tsA SV40 did not induce either gene mutations or chromosome aberrations at the same early stage after infection. Presumably the activity of the A gene of SV40 is necessary not only for the transforming but also for the mutagenic effect of the virus.

Topics: Animals; Cell Line; Cell Transformation, Viral; Chromosome Aberrations; Cricetinae; Cricetulus; Drug Resistance; Genes, Viral; Mercaptopurine; Mutation; Simian virus 40

Oncogenic bovine adenovirus (BAV3) was shown to induce chromosome aberrations and gene mutations to 6-mercaptopurine (6MP) resistance in Chinese hamster cells. BAV3 showed the highest mutagenic effect at the chromosome level 12--24 h postinfection. After 48 h the yield of aberrations dropped to the control level, where it remained after 72 and 96 h. BAV3 showed a highly significant induction of mutations to 6MP resistance 48 h postinfection. The effect of the combined treatment of cells with 5-bromodeoxyuridine (BrdU) and BAV3 on mutagenesis at chromosome and gene levels proved to be synergistic. The mutagenic activity at the gene level of BAV3, along with the earlier established mutagenic effect of SV40, indicates that this property is probably inherent in many DNA-containing oncogenic viruses. The possible mechanisms underlying malignant transformation and mutagenesis are discussed.

Topics: Adenoviridae; Animals; Cell Line; Cell Transformation, Viral; Chromosome Aberrations; Cricetinae; Cricetulus; Drug Resistance; Mercaptopurine; Mutation; Oncogenic Viruses