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No increases in chromosome aberrations in human diploid fibroblasts following exposure to low concentrations of sodium fluoride for long times.

To study whether exposure to fluoride at low concentrations for long times induces chromosome aberrations in human cells, human diploid fibroblasts in the quiescent phase were treated with sodium fluoride (NaF) at 1-10 micrograms/ml (equivalent to fluoride ion at 0.45-4.5 ppm) for 1-3 weeks. Quiescent cells were obtained by a 10-day culture in medium containing 1% serum following overnight incubation of cells in the logarithmic phase. Significant levels of cytotoxicity, as determined by a decrease in the number of cells, were not induced by treatment of the cells with NaF at 5 or 10 micrograms/ml for 1-3 weeks. No increase in the frequency of chromosome aberrations was elicited in cultures treated for 1-3 weeks with NaF over the range of doses examined. In contrast, a dose-dependent increase in the frequency of chromosome aberrations was observed in cultures treated with N-methyl-N'-nitro-N-nitrosoguanidine, used as a positive control. The results indicate that fluoride might be not clastogenic to human fibroblasts when exposed at low levels, equivalent to those in the communal water supplies.

Cells, Cultured↗

Retinoids induce sister-chromatid exchanges in human diploid fibroblasts.

Vitamin A palmitate, retinoic acid and an aromatic retinoic acid analogue (Ro 10-9359) induced sister-chromatid exchanges (SCE) in human diploid fibroblasts. The same SCE level was reached with the 3 compounds tested, when the concentration of vitamin A palmitate was 10 times higher than those of retinoic acid and Ro 10-9395. No correlation was found when the dose-related SCE induction of a retinoid was compared with the respective antineoplastic activity as reported by others.

Chromosome Aberrations↗

Testing of some permitted food colours for the induction of gene conversion in diploid yeast.

12 permitted food colours in use were screened for geno-toxicity. Mitotic gene conversion in Saccharomyces cerevisiae was used as the end-point. Each food colour was tested in stationary-phase as well as log-phase cells but without microsomal activation. These food colours did not cause any increase in mitotic gene conversion in diploid yeast BZ 34.

Drug Evaluation, Preclinical↗

Sister-chromatid exchange induction by metabolically activated retinoids in human diploid fibroblast cultures.

13-cis-Retinoic acid, retinyl-mthyl-ether, retinyl-phenyl-ether, retinyl-thio-ether and axerophthene each induced dose-dependent sister-chromatid exchanges (SCE) in human diploid fibroblasts. The functional relationship between retinoid concentration and SCE rate was similar in each of the 5 retinoids tested. The relationship reached a plateau at concentrations exceeding 8 micrograms/ml. alpha-Naphthoflavone (ANF), an inhibitor of P448-dependent mono-oxygenase, prevented the retinoid-induced increase of the SCE rate, but had no inhibitory effect in the presence of 4-nitroquinoline-1-oxide, an ultimate carcinogen. ANF did not reduce the spontaneously increased SCE rate in fibroblasts of patients with Bloom's syndrome. Retinoids failed to induce SCE in V79 Chinese hamster cells, which lack mono-oxygenase. Thus, we conclude that the retinoid-induced SCE rate increases independently of structural changes in the molecular side-chain ad that a metabolic activation of retinoids is required for SCE induction by cytochrome P448-dependent mono-oxygenase.

Abnormalities, Multiple↗

Genotoxicity of selenite in diploid yeast.

Selenite, a chemical of industrial importance and also an antimutagenic/anticarcinogenic agent, was tested for mutagenic and recombinogenic effects in 2 diploid yeast strains, Saccharomyces cerevisiae BZ 34 and D7. Selenite induced gene conversion and toxicity in BZ 34 and a variety of genetic events, viz. back-mutation, gene conversion, mitotic crossing-over, aberrant colony formation and also toxicity in the D7 strain. In both strains, the genetic effects of selenite showed a peak and a decline during 5 h of treatment while its toxicity increased marginally during 1-5 h. In the BZ 34 strain, the presence of glutathione (GSH) during selenite treatment greatly enhanced the convertogenic and toxic effects of selenite.

Chemical Phenomena↗

Botran and bleomycin induce crossing-over, and bleomycin also increases aneuploidy in diploid strains of Aspergillus.

Both bleomycin, an antineoplastic drug, and botran (2,6-dichloro-4-nitro-aniline), a fungicide, are known to inhibit growth and induce genetic segregation in diploid tester strains of Aspergillus nidulans when present in agar media. To identify primary effects, samples of induced apparent crossover types were analysed in detail. For both compounds, coincident and consecutive events of mitotic crossing-over were found to be very frequent and such events showed a random distribution among isolated colour sectors. In the case of botran, a thorough search for imbalanced precursor types was negative and recessive lethals were found only rarely. Such segregants are therefore unlikely the result of terminal deletions. For bleomycin, induction of reciprocal crossing-over was confirmed by treatments of germinating conidia. On plating to normal growth medium, crossover segregants showed up as coloured half- or quarter-colonies, including some "twin spots". Whole coloured colonies were also frequent and these increased with dose levels which caused decreasing survival and increasing frequencies of abnormal colonies. Analysis of large fractions of such "abnormals" identified aneuploids in all cases. While botran, in plate tests, also increased haploid segregants and disomic precursors could be found, tests of germinating conidia "in liquid" were inconclusive, because botran is insoluble in water. Some increases of aneuploids were observed, but only when botran and the solvent DMSO both were present at increased levels.

Aneuploidy↗

Intercellular karyotypic similarity in near-diploid cell lines of human tumor origins.

Intercellular karyotypic compositions were studied in seven near-diploid cultured cell populations (including six cell lines) of human colorectal and fibrosarcoma origins. Within each population, 50-88% of cells had the same cell-line-specific karyotype. In five of these seven populations, coexisting minor cell types could readily be identified, implicating the frequent occurrence of mosaicism in transformed cell lines. Nearly 90% of these cell types showed a single chromosome loss or gain from the modal cell karyotype. Parameters that may assist in the evaluation of karyotypic diversity of a transformed cell line are discussed briefly.

Adenocarcinoma↗

Cytogenetic analysis in human breast carcinoma. I. Nine cases in the diploid range investigated using direct preparations.

Cells in mitosis were found in 51 of 110 (47%) breast tumor samples; karyotypes of nine tumors in the diploid range are presented. The simplest stemline karyotype found was 46,XX, -16, +del(1)(qter----p21). The chromosome homologues most frequently lost were #8, #13, and #16. Monosomy or partial monosomy for chromosome #16 was seen in six cases, including the two simplest and chromosome #16 might be of relevance for initiation of malignant transformation in breast carcinoma. The only chromosome feature common to all nine breast carcinomas was the presence of a marker involving the long arm of chromosome #1, the region shared by all being 1qter----1q21.

Adult↗

Characteristic chromosomal imbalances in 18 near-diploid colorectal tumors.

The cytogenetic study of 18 near-diploid colorectal tumors shows that the observed numerical and structural abnormalities resulted in recurrent chromosomal losses and gains. By order of decreasing frequencies, they are: monosomy 17p (16/18), partial or more frequently complete monosomy 18 (14/18), trisomy 20q (11/18), trisomy or tetrasomy 13 (10/18), monosomy lp and trisomies X and 8q (9/18). The absence of recurrent breakpoints in euchromatin contrasts with the high preponderance of breakage at various places of heterochromatic region. Because these tumors are characterized by very recurrent chromosomal imbalances, it is assumed that the observed chromosomal changes may be related to a recessive genetic determinism and to gene dosage imbalances.

Adult↗

SK-UT-1B, a human tumorigenic diploid cell line.

SK-UT-1B (ATCC HTB 115) produced tumors 2 weeks after subcutaneous injection of 10 million cells into athymic nude mice. Karyotypic analysis at the 500-600 subband level, however, did not reveal any noticeable aberrations. This cell line therefore may represent the first example of experimentally proven human tumorigenic cells of solid tumor origin that retains the diploid karyotype in vitro indefinitely.

Aged↗

Existence of two distinct processes of chromosomal evolution in near-diploid colorectal tumors.

The comparison of all the karyotypes established in each of 18 near-diploid colorectal tumors made it possible to reconstruct a clonal evolution and to distinguish between early and late chromosomal aberrations. Because no abnormalities were observed in all tumors, and as even the most frequent changes, i.e., monosomy 17p and monosomy 18, may be present in mosaic, no chromosomal change can be regarded as a common primary event in the carcinogenetic process. However, the repeated occurrence of several changes favors the hypothesis of two karyotypic evolutionary processes. In most tumors, monosomy 17p and 18 were found, and the karyotypic evolution involved mainly several additional monosomies due to unbalanced rearrangements or losses that affect, by order of decreasing frequency, chromosomes 1p, 4, 14, 5q, 6q, 2p, and 11q, as well as gains of chromosomes 20, 8q, 13, 17q, and X. In this group of tumors, the mean number of chromosomes remains close to 46. In the other tumors, either only a monosomy 17p or a monosomy 18 was found and the karyotypic evolution involved essentially trisomies, resulting from gains with, by order of decreasing frequency, a preferential involvement of chromosomes 7, 8q, 13, 17q, 20, X, 2p, 5, and 16, the only additional recurrent deletion affecting chromosome 1p. In these tumors, the mean chromosome number is close to 51. Ten out of 11 polyploid sidelines emerged from monosomic-type tumors.

Chromosome Aberrations↗

Genetic analysis of repeated, biparental, diploid, hydatidiform moles.

A woman presented with five consecutive pregnancies displaying molar morphology. In the fifth pregnancy, a non-malformed, liveborn infant was delivered. Genetic analyses (RFLP analysis, cytogenetics, flow cytometry) were performed in pregnancies II-V. It was demonstrated that these pregnancies originated in separate conceptions, all conceptuses were diploid, and all had maternally as well as paternally derived genetic markers. By cytogenetic analysis, aberrant heteromorphisms were noted; no other abnormalities were observed in chromosome structure or in DNA sequence. Many different causes for the abnormal development can be envisaged, environmental as well as genetic. To conform to current ideas of molar pathogenesis, it is suggested that the present conceptuses might have arisen from imbalances in imprinted genomic regions. This could be a consequence of uniparental disomy in critical regions generated by somatic crossing over. Alternatively, it could be caused by a malfunction in the generation or maintenance of imprinting.

Chromosome Aberrations↗

Near-diploid karyotypes with recurrent chromosome abnormalities characterize early-stage endometrial cancer.

Cytogenetic investigation was attempted on 15 endometrial tumors. Whenever possible, a combination of direct harvesting and short-term culture (with or without prior methotrexate synchronization) was used. The analysis was successful in 13 cases: 12 carcinomas of stage I and one atypical hyperplasia. Clonal abnormalities were found in 10 tumors, whereas the remaining three showed a normal karyotype. The modal chromosome number was near-diploid. The abnormal karyotypes contained relatively simple numerical or structural aberrations in all but one tumor, a serous papillary carcinoma with multiple complex changes as well as cytogenetic evidence of intratumor heterogeneity. Gain of 1q, trisomy for chromosomes 2, 7, 10 (this trisomy was shown by in situ hybridization to be present also in a large number of interphase cells), and 12, and loss of chromosome 22 were recurrent aberrations; these are also the cytogenetic anomalies that have been consistently associated with endometrial carcinomas in previous studies. The utilization of both direct harvesting and short-term culture in several cases increased the frequency with which abnormal karyotypes were found; sometimes aberrations were found by the first method but not by the other, and vice versa. Never were different clonal anomalies found by the two approaches in the same case. Synchronization of the cultures generally led to chromosome preparations with more mitoses and of better quality. Again, no different anomalies were found in synchronized and standard cultures from the same tumor.

Chromosome Aberrations↗

Nonrandom numerical aberrations of chromosomes 7, 9, and 10 in DNA-diploid bladder cancer.

Double-target in situ hybridization technique (ISH) was applied to 37 cases of bladder cancer to detect numerical aberrations of chromosomes 7, 9, 10, and 11 by centromeric DNA probes. Of 33 evaluable cases, 29 (88%) demonstrated chromosome aberrations. In 17 cases with diploid pattern as measured by flow-cytometric DNA analysis (FCM), 15 (88%) demonstrated chromosome aberrations. Of these, trisomy 7, monosomy 9, and trisomy 10 were detected in three, three, and one case, respectively, as a single chromosome aberration. In 14 (88%) of 16 cases with an aneuploid DNA pattern, chromosome aberrations for two or more chromosomes were demonstrated. A significant correlation was observed between grade of chromosome aberrations and tumor grade (p < 0.01, Fisher's test), between number of spots for chromosome 7 and peak value in FCM (p = 0.015, by Spearman rank order test). In eight cases, chromosome aberrations in the tumor were compared with the corresponding pattern in the grossly normal and histologically benign mucosa. Trisomy 10 and monosomy 9 were detected as chromosome numerical aberrations in the histologically normal mucosa, consistent with aberrations in the corresponding patients. We conclude that trisomy 7, monosomy 9, and trisomy 10 may be early events in the evolution of bladder cancer.

Carcinoma, Transitional Cell↗

Cytogenetics of 158 patients with regional or disseminated melanoma. Subset analysis of near-diploid and simple karyotypes.

We report on the cytogenetic analyses of 158 cases of metastatic malignant melanoma, comprised of 63 cases with regional disease (RD) and 95 cases with distant (metastatic) disease (DD). Clonal structural abnormalities were identified in 126 (80%) cases and were significantly increased ( < 0.01 after adjusting for multiple comparisons) on chromosomes (in order of frequency of involvement) 1, 6, 7, 11, 9, and 3. Clustering of breakpoints occurred at 1p36, 1p22-q21, 6p11-q21, 9p, 11q23-qter, 13p (especially for cases with DD), and 19q13. The most common clonal numerical abnormalities, in a subset of 49 near-diploid cases were -10, -22, -9, +7, -19, and -Y. Analysis of chromosome segment gains and losses (CSRP) showed frequent loss of chromosomes 6 and 10, followed by equal rates of involvement of chromosomes 1, 7, and 9. Whole or segmental losses of chromosome 9 (especially 9p) correlate well with recent molecular genetic studies identifying putative suppressor genes, and are also likely important genetic abnormalities. However, based on the frequency of abnormalities in this large series of metastatic melanomas, it is likely that structural abnormalities of 1 and 6, and 10 are important in the pathogenesis of sporadic advanced melanoma.

Adult↗

Aneuploidy induction by mitotic arrestants. Effects of diazepam on diploid Chinese hamster cells.

We used diazepam, a mild mitotic arrestant, as a model chemical to develop a cytogenetic assay protocol for aneuploid induction in a diploid Chinese hamster cell system. At 100 micrograms/ml, diazepam almost completely arrested mitosis at metaphase. However, mitotic block could be quickly reversed when diazepam was removed from the medium. When the duration of arrest was relatively short (2 h), anaphases in the recovering cell population were essentially normal; but when the arresting period was prolonged (7 h), anaphases in the recovering cell populations exhibited a high rate of multipolar spindles and lagging chromosomes. Filial cell populations (18-24 h after removal of the drug) showed a high frequency of aneuploid elements. Mitotic cells shaken off from monolayer cultures as starting cell populations for enumeration of aneuploidy proved to be superior to non-shaken-off cell populations.

Anaphase↗

Combined mutagenic action of chemicals and radiation in diploid yeast.

The interaction between the mutagenic action of chemicals and radiation was studied by using a diploid yeast strain Saccharomyces cerevisiae BZ 34 with mitotic gene conversion as the end-point. The cells were treated with EMS, MMS or 4-NQO alone or in combination with gamma-radiation. The 2 alkylating agents EMS and MMS produced an additive mutagenic response, whereas 4-NQO exhibited an antagonistic effect in the combined treatment with gamma-radiation.

4-Nitroquinoline-1-oxide↗

Drug synergism or antagonism in the induction of diploid meiotic products in Saccharomyces cerevisiae.

The induction of yeast diploid meiotic products by treatment with bleomycin and mitomycin C is reduced when sporulating cells are treated in combination with propranolol, and increased when they are treated in combination with caffeine. We show that bleomycin and mitomycin C act by blocking the second meiotic division. The frequency of this event appears to be directly related to the intracellular cAMP concentration which is known to be influenced in opposite ways by caffeine and propranolol.

Antibiotics, Antineoplastic↗