Mitomycin C-induced Sister Chromatid Exchanges Research Articles

Broccoli flower head extract reduces mitomycin-C induced sister chromatid exchange in cultured human lymphocytes

This study is a continuation of our previous work [Murugan, S.S., Balakrishnamurthy, P., Mathew, Y.J., 2007. Antimutagenic effect of broccoli flower head by the Ames Salmonella reverse mutation assay. Phytother. Res. 21, 545–547], in search of possible antimutagenic properties in broccoli flower head extracts. In the present investigation, the effect of addition of ethanol extract of broccoli flower head on mitomycin-C (MMC) induced sister chromatid exchange (SCE) in cultured human peripheral blood lymphocytes was investigated. Broccoli flower head was extracted in ethanol using either acetone or ethanol as solvents. The extract was tested at final concentrations of 200 and 400 μg/ml culture and set for SCE assay. MMC at a concentration of 1 μg/ml and the test concentrations of broccoli flower head were added to the culture following 48 h from the initiation of culture. Enumeration of SCE in second division mitotic cells indicated that broccoli flower head extract significantly reduced MMC induced SCEs at both the concentrations tested. This observation is in line with our earlier finding and confirms to the presence of antimutagenic principles in broccoli flower head extract.

Mitomycin C-induced sister chromatid exchanges (SCE) in patients undergoing hyperbaric oxygen therapy

Mitomycin C-induced sister chromatid exchanges (SCE) in patients undergoing hyperbaric oxygen therapy

Cytogenetic Effects of Grape Extracts (Vitis vinifera) and Polyphenols on Mitomycin C-Induced Sister Chromatid Exchanges (SCEs) in Human Blood Lymphocytes

In the present study, the effects of extracts and polyphenol-rich fractions as well as monomer polyphenols identified in them, from both red and white grapes, on mitomycin C (MMC) induced sister chromatid exchanges (SCEs) in human peripheral blood lymphocytes were investigated. The grape extracts and two of the three polyphenol-rich fractions promoted MMC-induced SCEs at concentrations from 75 to 300 microg/mL. However, none of the extracts or fractions alone induced SCEs. Thus, these results suggest caution especially with regard to the use of grape extracts as dietary supplements. On the other hand, the fact that these extracts were not genotoxic alone may indicate a selective activity against genetically damaged cells. This is the first study regarding the clastogenic effects of grape extracts in human cells. Moreover, from the tested polyphenols, caffeic acid, gallic acid, and rutin hydrate enhanced MMC-induced clastogenicity, whereas ferulic acid, protocatechuic acid, (+)-catechin, (-)-epicatechin, and trans-resveratrol had no effect at concentrations between 5 and 100 microM. The differences in the chemical structures of the tested polyphenols may account for their differential effects on MMC clastogenicity.

Biochemical Analysis of the DNA Unwinding and Strand Annealing Activities Catalyzed by Human RECQ1*[boxs

RecQ helicases play an important role in preserving genomic integrity, and their cellular roles in DNA repair, recombination, and replication have been of considerable interest. Of the five human RecQ helicases identified, three are associated with genetic disorders characterized by an elevated incidence of cancer or premature aging: Werner syndrome, Bloom syndrome, and Rothmund-Thomson syndrome. Although the biochemical properties and protein interactions of the WRN and BLM helicases defective in Werner syndrome and Bloom syndrome, respectively, have been extensively investigated, less information is available concerning the functions of the other human RecQ helicases. We have focused our attention on human RECQ1, a DNA helicase whose cellular functions remain largely uncharacterized. In this work, we have characterized the DNA substrate specificity and optimal cofactor requirements for efficient RECQ1-catalyzed DNA unwinding and determined that RECQ1 has certain properties that are distinct from those of other RecQ helicases. RECQ1 stably bound to a variety of DNA structures, enabling it to unwind a diverse set of DNA substrates. In addition to its DNA binding and helicase activities, RECQ1 catalyzed efficient strand annealing between complementary single-stranded DNA molecules. The ability of RECQ1 to promote strand annealing was modulated by ATP binding, which induced a conformational change in the protein. The enzymatic properties of the RECQ1 helicase and strand annealing activities are discussed in the context of proposed cellular DNA metabolic pathways that are important in the maintenance of genomic stability.

Rad51C-deficient CL-V4B cells exhibit normal levels of mitomycin C-induced SCEs but reduced levels of UVC-induced SCEs

The mechanisms of sister chromatid exchanges (SCEs) are not known. One hypothesis is that SCE is a manifestation of Rad51-dependent homologous recombination repair. In order to test this hypothesis, we have compared the frequencies of SCEs induced by mitomycin C (MMC) and 254 nm ultraviolet radiation (UVC) in wt V79B and the Rad51C-deficient CL-V4B cells. SCEs were analysed in the first (M1) and second (M2) post-treatment mitoses. In M1 MMC induced the same frequencies of SCEs in CL-V4B and V79B cells, while the UVC-induced SCE frequencies were lower in CL-V4B than V79B cells. In CL-V4B cells, MMC-induced SCEs were higher in M2 than in M1, suggesting that interstrand cross-links (ICL) are either not removed completely or are transformed into another form of DNA damage that persists until the next cell cycle. We suggest that SCEs may represent a mechanism to bypass MMC-induced ICL without their removal.

The association between frequencies of mitomycin C-induced sister chromatid exchange and cancer risk in arseniasis

In order to examine whether biomarkers of cytogenetic damage and susceptibility, such as spontaneous and mitomycin C-induced sister chromatid exchange (SCE) can predict cancer development, a nested case-control study was performed in a blackfoot endemic area with known high cancer risk. A cohort of 686 residents was recruited from three villages in the arseniasis area. Personal characteristics were collected and venous blood was drawn for lymphocyte culture and stored in a refrigerator. The vital status and cancer development was followed using the National Death Registry, Cancer Registry, and Blackfoot Disease Registry. The follow up period was from August 1991 to July 1997. During this 6-year-period, 55 residents developed various types of cancer. Blood culture samples from 23 of these subjects were unsuitable for spontaneous SCE experiments and 45 of these subjects were unsuitable for mitomycin C-induced SCE experiments due to improper storage. Finally, a total of 32 cancer cases had cytogenetic samples that could be analyzed. About 32 control subjects were selected from those who did not develop cancer in the study period and these subjects were matched to cases by sex, age, smoking habits, and residential area. The results showed that there was no significant difference in the frequencies of spontaneous and mitomycin C-induced SCE between the case and control groups. There was also no significant difference in the net difference of spontaneous and mitomycin C-induced SCE between the case and control groups. These results suggest that SCEs, either spontaneous or mitomycin C-induced, might not be good markers to predict cancer risk.

Inhibition of mitomycin C-induced sister chromatid exchanges by vitamins C, E and beta-carotene

Inhibition of mitomycin C-induced sister chromatid exchanges by vitamins C, E and beta-carotene

Modulation of mitomycin C-induced sister chromatid exchanges and cell cycle delay by buthionine sulfoximine and reduced glutathione in mouse bone marrow cells in vivo

Mitomycin C (MMC) is a bifunctional alkylating agent and a chemotherapeutic drug, covalently binds to the DNA of cells and produces monoadducts and DNA–DNA, DNA–protein crosslinks, induces damages at chromosomal level and slows down the rate of cell proliferation. Reduced glutathione (GSH), a major non-protein thiol substance plays an important role in detoxification of cells against the toxic effect of exogenous compounds. In order to understand the role of factor which affects MMC sensitivity, we have made an attempt to establish a relationship between MMC-induced DNA damages and the endogenous GSH-status of the cells. MMC was treated to normal and buthionine sulfoximine (BSO; GSH-depleting agent)-treated mice. Exogenous GSH was also added to MMC-treated normal mice. Cells were fixed at 24 h and sister chromatid exchanges (SCEs) and average generation time (AGT) were scored. MMC-induced SCEs and cell cycle delay significantly with respect to control and the frequency of SCEs was increased considerably while MMC treatment combined with either GSH or BSO. The induction of cell cycle delay by MMC was reduced significantly when GSH or BSO was present along with MMC. These observations indicate that the factor responsible for inducing delay in cell cycle after MMC treatment may not be relevant for SCE-induction.

Suppression of the frequencies of baseline and mitomycin C-induced sister chromatid exchanges by ginsenoside Rh 2 in human blood lymphocytes

Suppression of the frequencies of baseline and mitomycin C-induced sister chromatid exchanges by ginsenoside Rh 2 in human blood lymphocytes

In vitro effects of prostaglandin E1 and indomethacin on mitomycin C-induced sister-chromatid exchanges in mitogen-stimulated human lymphocytes

In this study, the individual and combined effects of prostaglandin E1 (PGE1) and Indomethacin on mitomycin C (MMC)-induced SCEs in human lymphocytes was investigated in vitro. All MMC-treated cultures showed a great increase of SCEs (approximately two-fold), indicating its ability to induce mutations. SCE data showed that MMC-induced SCEs were reduced significantly in the presence of PGE1 in pooled analysis of six experiments (60.55% reduction of SCEs at 10 −6 M, 34.13% reduction of SCEs at 10 −9 M). In contrast the presence of indomethacin in the medium during MMC treatment of cells failed to show a significant reduction of SCEs in pooled analysis (21.17%). However, individual analyses revealed only two of six donors with a significant SCE response. Thus, the findings suggest that PGE1 can modify the DNA damaging effect of carcinogens and thereby may prevent the initiation of the carcinogenic process.

Mitomycin C-Induced Sister Chromatid Exchange in X Chromosomes of Bovidae

Sister chromatid exchanges (SCEs) in early- and late-replicating X chromosomes of seven female cattle (Bos taurus L.) and five female river buffalo (Bubalus bubalis L.) were studied in untreated lymphocytes and lymphocytes treated with mitomycin C (MMC). In the experiment, 577 SCEs on X chromosomes of MMC-untreated cells and 825 SCEs on X chromosomes of MMC-treated cells from both species were observed. No significant differences between the number of SCEs in early- and late-replicating X chromosomes were found even when singular species and subjects were considered.

Sister-chromatid exchanges induced by mitomycin C after acute or chronic exposure of human lymphocytes to a low dose of X-rays

Human peripheral blood lymphocytes from 10 male donors were exposed to mitomycin C with and without prior irradiation with 0.01 Gy X-rays. Acute or chronic irradiation of lymphocytes in G 1 resulted in a decrease in the subsequent level of mitomycin C-induced sister-chromatid exchange aberrations. The effect was small (∼ 10%) with evidence of some variability between donors. By contrast no adaptive response was observed if the cells were treated in G 0.

Increased spontaneous and mitomycin C-induced sister chromatid exchanges in patients with cancer of the cervix uteri, with special reference to stage of cancer

The frequencies of spontaneous and mitomycin C (MMC)-induced sister chromatid exchange (SCE) were examined in 35 patients with cancer of the cervix uteri (stage 0, eight cases; stage I, nine cases; stage II, nine cases, and stage III, nine cases) before they had undergone cancer treatment, as well as in seven patients with uterine myoma and 18 healthy women as controls. The frequency of SCE was analyzed in reference to the stage of cancer in the cancer group and in reference to chromosome group in the cancer and normal groups. The frequencies of spontaneous and MMC-induced SCE in the cancer group were 10.0 ± 1.8 and 20.7 ± 2.6, respectively, and both were significantly higher than in the myoma (8.1 ± 0.8 and 17.6 ± 1.8) and normal (7.6 ± 0.8 and 17.6 ± 2.3) groups. Furthermore, the frequency of SCE in the cancer group increased with cancer stage. All chromosome groups contributed equally to the increase in SCE in the cancer group. These results indicate that an increase in the frequency of SCE in patients with cervical cancer is related to the presence of cancer, but is not related to a predisposition to cancer.

Inhibition of mitomycin C-induced sister-chromatid exchanges in mouse bone marrow cells by the immunopotentiators Krestin and Lentinan

We have carried out an investigation to determine whether or not the sister-chromatid exchange frequencies (SCEs) observed in bone marrow cells in mice treated with mitomycin C (MMC) are inhibited by the immunopotentiators Krestin and Lentinan. We found that mitomycin C (2 mg/kg, i.v.)-induced SCEs were inhibited in 27% of the mice treated with Krestin (300 mg/kg, i.p.) and in 23% of the mice treated with Lentinan (1 mg/kg, i.p.). The effects of Krestin were found to be dose-dependent in inhibition of MMC-induced SCEs while those of Lentinan were not. Our findings therefore suggest that Krestin and Lentinan are not only useful for cancer treatment as immunopotentiators in combination with anticancer drugs but may also prevent the increase of chromosomal damage induced by anticancer drugs.

Sister chromatid exchange frequency in a retinoblastoma mosaic patient with del(13)

The frequencies of sister chromatid exchange (SCE) were investigated in different cell populations derived from a patient with retinoblastoma and 46,XY/46,XY,del(13) (q12.3q21.2) mosaicism. No differences in spontaneous or mitomycin C-induced SCE were detected between cell populations.

Enhancement of cis-diamminedichloroplatinum and mitomycin C-induced sister chromatid exchanges by dibutyryl cyclic AMP in a human salivary gland adenocarcinoma cell line in culture.

A human salivary gland adenocarcinoma cell line was cultivated in the presence of dibutyryl cyclic AMP (dB-cAMP), cis-diammine dichloroplatinum (cisplatin) or mitomycin C (MMC) only, or of the combination of dB-cAMP and each of the antineoplastic drugs. Then the treated cells were examined for the induction of sister chromatid exchanges (SCEs), colony-forming efficiency (CFE) in semisolid agar medium and cell survival as measured by counting for viable cells. It has been found that the frequency of SCE induction is significantly increased by treatment of cells with cisplatin or MMC under the presence of dB-cAMP, when compared to that in the cells treated with cisplatin or MMC only. Moreover, marked reduction in CFE and cell survival was found to occur in the cells treated with the combination of dB-cAMP and cisplatin or MMC. These finding indicate that treatment of a human salivary gland adenocarcinoma cell line with dB-cAMP results in modification of SCE induction, CFE and cell survival in the cells exposed to cisplatin or MMC.

Chromosome studies in inherited nonpolyposis colon cancer syndrome

This study was designed to determine if any constitutional chromosomal markers were linked with the expression of colorectal neoplasms in the inherited nonpolyposis colon cancer syndrome, using a number of cytogenetic techniques. High resolution G-banding in 12 affected and 17 unaffected family members did not reveal a structural chromosome abnormality. Increased C-band heteromorphism was not seen in either affected or unaffected individuals, and no heritable fragile sites were detected. Mean baseline and mitomycin C-induced sister chromatid exchanges were not elevated in affected patients compared with controls. Mapping of sister chromatid exchanges did not reveal any hot spots of exchange. A tumor cell line with the karyotype 46,XY,der(13),t(13;?)(p11;?) was established from one patient, but no constitutional abnormality of chromosome #13 was found. In addition, 11 patients with familial polyposis coli were studied with high resolution G-banding and no heteromorphism of chromosome #2 in the region 2q21.3 was detected.

Spontaneous and induced sister chromatid exchanges in lymphocytes of healthy persons

Spontaneous and mitomycin C-induced sister chromatid exchanges (SCEs) in lymphocytes were analyzed in 46 healthy persons. Our results suggest that the frequency of spontaneous SCE in lymphocytes is useful for evaluation of the biological effects of environmental mutagens, and that the induced-SCE test with a mutagen and lymphocytes may be used to detect persons with a high sensitivity to the mutagen.

Effects of smoking on spontaneous and induced sister chromatid exchanges in lymphocytes

Spontaneous and mitomycin C-induced sister chromatid exchanges (SCEs) in lymphocytes were analyzed in 24 non-smokers and 24 sex- and age-matched smokers. Mean spontaneous SCE frequency for non-smokers was 9.8 SCEs/cell, and that for smokers was 11.5 SCEs/cell. The difference was statistically significant ( P < 0.001 by t-test). These results suggest that spontaneous SCE frequency in lymphocytes is useful for evaluation of biological effects of environmental mutagens. However, we could not find any effects of smoking on the sensivities of lymphocytes to mitomycin C in vitro. The effects of mutagens on humans may be independent of one another.

Cytogenetic and cytokinetic analysis of lymphocytes from patients with hereditary adenomatosis of the colon and rectum

The frequencies of chromosome aberrations, sister chromatid exchanges (SCEs), and cell cycle kinetics were examined in cultured lymphocytes from five patients with hereditary adenomatosis of the colon and rectum (ACR) [three patients with Gardner's syndrome (GS) and two patients with familial polyposis coli (FPC)]. The frequency of numerical chromosome aberrations was no different in metaphase cells at the first and second replication cycles (M1 and M2) in the ACR patients and control subjects. The percentage of structural chromosome aberrations in both M1 and M2 cells was somewhat higher in the ACR patients as compared with the controls. Neither spontaneous nor mitomycin C-induced SCE frequencies in the patients with ACR were different from the controls, except for one patient with GS, who showed a remarkably high spontaneous SCE frequency. This patient is the mother of a son who had hepatoblastoma. The cell replication index (RI) was lower in the GS patients than in the controls. However, the RI in the FPC patients did not differ from that of the controls.