[en] The frequency of ultraviolet light induced neoplastic transformation of Syrian hamster embryo cells is enhanced 3- to 10-fold when the cells are first treated with either x-irradiation or with methyl methanesulfonate. Maximum enhancement occurs when the interval between the two treatments is 48 h. The relevance of uv-induced transformation to neoplasia is confirmed because the transformation to neoplasia is confirmed because the transformants produce tumors when injected into nude mice. Excision and postreplication DNA repair were studied to determine whether the enhanced transformations were associated with either of these repair mechanisms. Independent of x-ray or of methyl methanesulfonate pretreatment, approximately 25% of the pyrimidine dimers are excised within 24 h in cells irradiated with uv with 3 J/sq m. During this period, more than 70% of the genome of cells irradiated with uv has been replicated. Postreplication repair is measured by the time required to chase pulse-labeled nascent DNA strands to parental-sided DNA. Regardless of pretreatment, 1 and 3 h are required for pulse-labeled DNA in control and irradiated cells, respectively, to reach parental size. Therefore, no correlation is found between a change in the rate of excision or postreplication repair and enhancement of transformation. Relative to control cloning efficiency, the survival of HEC contain more than 10(5) pyrimidine dimers/genome. The level of survival is similar to the survival of human skin fibroblasts which excise pyrimidine dimers four to five times as efficiently. Moreover, postreplication repair cannot account for the ability of these cells to survive because it is three times slower than in human fibroblasts. Therefore, other repair mechanisms must be responsible for HEC survival and transformation

[en] The development of reproducible quantitative in vitro procedures resulting in neoplastic transformation of mammalian cells has made possible the separation of events related to the process leading to transformation from secondary events that interfere with the early recognition of transformation. The use of chemical carcinogens on Syrian hamster cell strains results in a dose-response relation consistent with a Poisson distribution, indicating that the transformation phenomenon is inductive. In some circumstances, the joint action or interaction of chemical carcinogens with other agents results in an increased incidence of transformation. The pretreatment of Syrian hamster cells with ionizing radiation (250 R) or alkylating chemicals enhances the frequency of transformation on a cell or colony basis ordinarily obtained with known chemical carcinogens. Pretreatment with non-ionizing irradiation (uv, 254 nm) did not have a similar effect. The two types of irradiation and the alkylating agents reduced the cloning efficiency of the cells. X ray alone produced no transformation; the alkylating chemicals produced transformations infrequently, whereas uv produced a significant number of transformations. The number of transformations associated with uv is increased by pretreatment of the cells by x-irradiation. The enhancement of transformation by x-ray or x-ray-type agents appears to be independent of the type of second carcinogen used

[en] The x-irradiation of cultured Syrian hamster cells followed by exposure to a chemical carcinogen after seeding for colony formation results in an enhancement of transformation ordinarily associated with the chemical. Maximum enhancement occurred when 48 hr separated the two treatments and when 250 R was used. With different concentrations of benzo(a)pyrene (0.1-5 μg/ml medium) a similar factor of enhancement was obtained with 250 R, which averaged 8.35, implying that the irradiation pretreatment caused a constant multiplying factor. These results are consistent with the interpretation that transformation occurs through interactions in keeping with a one-hit hypothesis. The transitory effect of irradiation and the lack of transformation with irradiation alone argues against the selection of a special radiation-sensitive cell. At the time of maximum enhancement (48 hr), the cell cycles of irradiated and nonirradiated cells do not differ. The addition of benzo(a)pyrene to irradiated cells does not alter the number of cells capable of synthesizing DNA or the flow of cells from G₁ to S. The number of cells accumulated in mitosis also does not appear to differ between irradiated and nonirradiated cultures treated with 2.5 μg benzo(a)pyrene. Chromosome aberrations or alterations in ploidy cannot account for the significant increase in enhancement observed at 48 hr postirradiation; however, twice as many cells with chromosome or chromatid gaps were found at 48 hr as compared to either 24 or 72 hr postirradiation

[en] The induction of neoplastic cell transformation is closely associated with DNA alterations which occur shortly after carcinogen exposure. Sister chromatid exchange (SCE) formation is a sensitive indicator of carcinogen-DNA interaction and correlates with the induction of morphological cell transformation. The persistence of lesions generating SCE produced by chemical and physical carcinogens and its relevance to the induction of morphologic transformation was evaluated in coordinated experiments with cultured Syrian hamster fetal cells (HFC). Exponentially growing HFC were exposed for 1 h to benzo[a]pyrene (BP), methyl-methanesulfonate (MMS), cis-platinum (II) diaminedichloride (cis Pt II), N-methyl-N'-nitrosourea (MNU), mitomycin C (MMC), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), N-acetoxy-2-fluorenyl-acetamide (AcAAF) or u.v. light irradiated. SCE analysis demonstrates that for a period of 48 h after carcinogen exposure, during which time the cells undergo at least four replicative cycles, DNA damage generating SCE induced by all chemical carcinogens either persisted or was partially removed, whereas u.v.-induced lesions were completely removed. An elevated SCE frequency persisted after two additional cell cycles after treatment with BP, AcAAF or MMC without increased cell lethality as compared to other carcinogens whose lesions were completely eliminated during the same period

[en] The nature of DNA replication in UV irradiated Syrian hamster embryo cells (HEC) was investigated by measuring the size distribution of nascent daughter strand DNA. During the early mode, nascent strands are made in smaller pieces than in nonirradiated cells. The late mode begins when nascent strands recover to normal size. This was observed in HEC 5 h post-UV. When the late mode is operational, nascent strands elongate to parental size in <2 h, whereas >3 h are required during early mode function. Evidence from split dose experiments demonstrates that the recovery of the size of nascent strands is not due to enhanced gap filling. Furthermore, pyrimidine dimers are probably recognized differently by the replication complex during early and late mode DNA synthesis. The late mode of replication could account for the ability of HEC to survive UV irradiation even though they are inefficient in both excision and postreplication repair

[en] U.V.-exposure of foetal hamster cells (secondary or tertiary sub-cultures) seeded for colony formation resulted in the induction of transformation. The U.V.-effect on colony-forming ability resulted in a D₃₇ of 58 ergs/mm². The observed transformation frequency induced by U.V.-increased approximately in proportion to increased doses in the range of 7.5 to 60.8 ergs/mm² examined. When cells seeded for colony formation were treated with U.V. and later with either benzo(a)pyrene or N-acetoxy-fluorenyl-acetamide, commencing 6 min after exposure to U.V., there was neither an additive nor a synergistic enhancement of the transformation frequency ordinarily observed with either chemical agent alone. In mass cultures that were X-irradiated with 250 R, seeded for colony formation and treated with either 15 or 30 ergs.mm² of U.V. 48 hours later, the enhancement of transformation was approximately 12- and 6-fold on a colony basis and 6- and 3-fold on a dish basis, respectively. No transformation occurred after X-irradiation only. The enhancement obtained by the pre-treatment with X-ray is similar to that reported for the combination of X-ray and chemical carcinogens. (author)

[en] The effect of caffeine on post-uv DNA replication was studied to determine its relevance to carcinogenesis. The level of uv-induced transformed colonies of Syrian hamster embryo cells (HEC) was increased up to fivefold when caffeine was added to cells between 0 and 6 h post-uv. The greatest increase was observed when the interval between uv irradiation and caffeine addition was 4 h. Two modes of DNA replication occurred after uv irradiation. During the early mode (0 to 3 h post-uv) the size of nascent strands, as measured by alkaline sucrose sedimentation, was smaller than those in nonirradiated cells, whereas during the late mode they recovered to normal size. Caffeine inhibited the rate of elongation of nascent strands during the early mode. When caffeine was added immediately after uv irradiation, the conversion of the early mode to the late mode was inhibited. Studies on the effects of caffeine have now been extended to the late mode. While caffeine has little effect with the fd elements beginning from the 10th day after irradiation is connected with their proliferation but not with the migration out from lymphoid organs

[en] Normal human foreskin keratinocytes cotransfected with the neomycin resistance gene and recombinant human papillomavirus (HPV) DNAs (types 16, 18, 31, and 33) that have a high or moderate association with cervical malignancy acquired immortality and contained integrated and transcriptionally active viral genomes. Only transcripts from the intact E6 and E7 genes were detected in at least one cell line, suggesting that one or both of these genes are responsible for immortalization. Recombinant HPV DNAs with low or no oncogenic potential for cervical cancer (HPV1a, -5, -6b, and -11) induced small G418-resistant colonies that senesced as did the nontransfected cells. These colonies contained only episomal virus DNA; therefore, integration of HPV sequences is important for immortalization of keratinocytes. This study suggests that the virus-encoded immortalization function contributes to the pathogenesis of cervical carcinoma

[en] Normal human foreskin fibroblasts treated in vitro with a chemical carcinogen or irradiated with ultraviolet light subsequently acquired anchorage independent growth and an extended but finite capacity for exponential growth. All cell lines were derived from cells recovered from colonies that had grown in semisolid medium; cell lines originally treated with a chemical carcinogen produced nodules after s.c. inoculation into nude mice. G-banding analysis of 10 cell lines (including one ultraviolet light line) revealed that seven were chromosomally abnormal with structural and numerical chromosome alterations, one was characterized by a consistent trisomy, and the other two were normal diploid. Structural alterations consisted of chromosome deletions, translocations, and partial chromosome duplications. Although no common structural or numerical abnormality was detected, several structural alterations were observed involving chromosomes 1, 7, 11, and 22, where fgr, erb-B, H-ras-1, and sis protooncogenes, respectively, are located. In one cell line trisomy 17 was a unique chromosome alteration. The induction of chromosome changes may have influenced the proliferative capacity of the treated cells relative to nontreated cells. However, the two cell lines without detectable chromosome changes also had an increased proliferative life span, suggesting that other submicroscopic genetic alterations may have affected cell multiplication. Although carcinogen induced chromosome abnormalities may represent a step in the process of neoplastic development, additional genetic and/or epigenetic changes, are required for indefinite growth and the expression of malignancy

[en] The influence of caffeine post-treatment on sister-chromatid exchange (SCE) and chromosomal aberration frequencies on Chinese hamster cells exposed to a variety of chemical and physical agents followed by bromodeoxyuridine (BrdUrd) was determined. (Auth.)