• Toxicological Research
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• v.25 no.4
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• pp.189-193
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• 2009

Hypoxia inducible factor $1\alpha$ (HIF-$1\alpha$) is a potential marker of carcicnogenesis since it is overexpresssed in many human cancers such as brain, breast, and uterus, and its role has implicated in tumor cell growth and metastasis. In this study, we established a stable cell line that express green fluorescence protein (GFP)-fused hypoxia inducible factor $1\alpha$ (HIF-$1\alpha$) and evaluated the potential use of this cell line for assessment of carcinogenicity of chemical toxicants. Western blot analysis as well as fluorescence measurements showed that protein-level of GFP-HIF-$1\alpha$ was significantly enhanced in a dose-dependent manner upon treatment of hypoxia mimicking agents such as dexferrioxamine and $CoCl_2$. Well-Known tumor promoters such as mitomycin and methyl methanesulfonate. significantly induced the fluorescence intensity of GFP-HIF-$1\alpha$, whereas the known negative controls such as o-anthranilic acid and benzethonium chloride, did not. These results indicate that HIF-$1\alpha$ could be a biological parameter for detection of tumor initiators/promoters and suggest that the GFP-HIF-$1\alpha$ cell line is a useful system for screening of carcinogenic toxicants.

• Environmental Mutagens and Carcinogens
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• v.16 no.1
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• pp.19-23
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• 1996

This study was performed by the sister chromatid exchanges (SCEs) and micronuclei (MN) assays to investigate the adaptive response to ultraviolet light (UV) or ethyl methanesulfonate (EMS) in Chinese hamster ovary (CHO) and Sarcoma 180 (S180) cells. The pretreatment with 1 J/m$^2$ UV or 2 mM EMS decreased the frequency of SCEs induced by the treatment with 5 J/m$^2$ UV or 8 mM EMS in CHO cells. The pretreatment with UV (1 or 2 J/m$^2$) or EMS (1, 2 or 3 mM) did not affect the SCEs induced by the treatment with 7 J/m$^2$ UV or 10 mM EMS in S180 cells. On the other hand, the pretreatment with 1 J/m$^2$ UV or 2 mM EMS decreased the frequency of MN induced by the treatment with 5 J/m$^2$ UV or 8 mM EMS in CHO cells. The pretreatment with UV (1 or 2 J/m$^2$) or EMS (1, 2 or 3 mM) did not affect the frequency of MN induced by the treatment with 7 J/m$^2$ UV or 10 mM EMS in S180 cells. It is suggested that there are adaptive responses at the level of chromosome and micronuclei to UV and EMS in CHO cells.

• The Korean Journal of Zoology
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• v.26 no.3
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• pp.171-179
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• 1983

Amounts of DNA single strand breaks and unscheduled DNA synthesis in CHO cells exposed to MMS were increased in the presence of 3-aminobenzamide, a potent inhibitor of poly (ADP-ribose) polymerase. However, those in cells irradiated with UV-light were decreased. These results suggest that poly (ADP-ribose) polymerase acts negatively on the MMS-induced base excision repair but positively on the UV-induced nucleotide excision repair. In the combined treatment with MMS and UV-light in the presence of this inhibitor, amounts of strand breaks were just the same as those in the absence of the inhibitor. But those of unscheduled DNA synthesis were increased up to the amount induced by UV-light alone. These results may suggest that poly (ADP-ribose) polymerase affects the incision step of excision repair induced by MMS and UV-light independently, and that it may potentiate the complete cleaving of UV-induced pyrimidine dimers possibly by the repair enzymes which might have been partially inactivated by MMS.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.50 no.6
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• pp.824-828
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• 2017

We investigated the anesthetic effects of MS-222 (tricaine methanesulfonate), clove oil, 2-phenoxyethanol, $NaHCO_3$, lidocaine-HCl and lidocaine-$HCl/NaHCO_3$ in the glass catfish Kryptopterus vitreolus. Based on the efficacy criteria of complete anesthetic induction from 60 s to 120 s, recovery within 300 s, the lowest effective concentrations at $24^{\circ}C$ were determined to be 60 ppm (induction $82.8{\pm}17.6s$, recovery $80.2{\pm}34.7s$) for MS-222, 40 ppm (induction $70.5{\pm}8.2s$, recovery $83.4{\pm}17.7s$) for clove oil, 250 ppm (induction $64.3{\pm}24.0s$, recovery $62.8{\pm}15.6s$) for 2-phenoxyethanol, 300 ppm (induction $127.3{\pm}13.3s$, recovery $107.5{\pm}4.8s$) for lidocaine-HCl and 200/100 ppm (induction $81.2{\pm}17.2s$, recovery $98.3{\pm}19.7s$) for lidocaine-$HCl/NaHCO_3$. Thus, 200/100 ppm of lidocaine-$HCl/NaHCO_3$ was found to be an effective anesthetic agent.

• Korean Journal of Microbiology
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• v.38 no.2
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• pp.96-102
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• 2002

The S-phase checkpoint mechanisms response to DNA damage or inhibition of DNA replication for maintenance of genetic stability in eukaryotic cells. These roles include cell cycle control arrest at S-phase and Iranscriptional induction of repair genes. To characterize the defects of dpbll mutant for both these responses, we examined the over-expression effect of DPBll gene, the sensitivity to HU, MMS, and the transcriptional pattern by DNA damage agent for RNRS mRNA. RNRS transcript is induced in response to a wide variety of agents that either damage D7A directly through chemical modification or induce stress by blocking DNA synthesis. As results, dpbll-1 cells are sensitive to DNA damage agents and the level of RNR3 mRNA is reduced approximately 40% than wild type cells. Moreover, we found the same results in dpb2-1 cells. Therefore, we propose that DPB2 and DPBll act as a sensor of replication that coordinates the transcriptional and cell cycle responses to replication blocks.

• Journal of Life Science
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• v.16 no.1
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• pp.126-130
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• 2006

The present study intends to characterize the DNA damage-inducible responses in yeast. The fission yeast, Schizosaccharomyces pombe was used in this study as a model system for higher eukaryotes. To study UV-inducible responses in S. pombe, five UV-inducible cDNA clones were isolated from S. pombe by using subtration hybridization method. To investigate the expression of isolated genes, UVI-155, the cellular levels of the transcripts were determined by Northern blot analysis after UV-irradiation. The transcripts of isolated gene (UVI-155) increased rapidly and reached maximum accumulation after UV-irradiation. Compared to the message levels of control, the levels of maximal increase were approximately 5 fold to UV-irradiation. In order to investigation whether the increase of UVI-l55 trascripts was a specific results of UV-irradiation, UVI-155 transcript levels were examined after treating the cells to mthylmethane sulfonate (MMS). The transcripts of UVI-155 were not induced by treatment of $0.25\%$ MMS. These results implied that the effects of damaging agents are complex and different regulatory pathways exist for the induction of these genes. To characterize the UVI-155 gene, gene deletion experiments were analyzed. The deleted strain was not well grown. This result indicated that the UVI-155 gene is essential for cell viability.

• Applied Microscopy
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• v.28 no.4
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• pp.447-463
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• 1998

This study was carried out to investigate the histomorphological changes and the electrophoretic patterns of egg components, obtained from 100 of 2-year-old female catfishes (Silurus asotus). Female Korean catfishes collected in the vicinity of Chollabukdo have synchronous ovaries which discontinously ovulate eggs during the breeding season (from late May until early July). The fishes were killed by severing the spinal cord just posterior to the head after immobilization with tricaine methanesulfonate (MS 222). Especially, the light microscopic and ultrastructural changes of ooplasm and follicular membranes of oocytes, were examined by means of light and transmission electron microscopy. The size of the nucleoli and number of the yolk granules increased as the oocyte groved. Yolk granules were deposited in the oocyte as fluid Due to tile presence of large early and late maturing oocytes, their ovaries were enlarged, transparent, granular and greenish in color. As the percentages of fish in late maturing oocyte (LMO) and ripe oocyte (RO) stage increased from March to April, mean gonadosomatic index (GSI) values (19.95%) increased. Zona radiata changed from squamous into cuboid in shape in the early maturing oocyte (EMO) stage. Processes, microvilli, from the zona radiata and from the oocyte grow, and make contact with each other in the pore canals of the zona radiata during vitellogenesis, but are withdrawn as the zona radials becomes more compact and devoid of pore canals during oocyte maturation. Seasonal changes in the microscopic appearance of the ovaries were well correlated with those in both gonadosomatic index and macroscopic appearance. The main cytological changes such as increase in size of cell, nucleus, nucleolus, and increase in number of nucleoli and mitochondria demonstrated with electron microscopy in the previtellogenic oocytes of Korean catfish, provided evidences for important synthetic processes in an early preparatory phase of oocyte development. The electrophoretic pattern of major band in mature stage was much thicker (24 k, 66 k, 90-110k dalton) than that in previtellogenic phase.

• Korean Journal of Plant Resources
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• v.34 no.2
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• pp.172-176
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• 2021

The Hosta 'Hwangnarae' was bred at the Korea National Arboretum, which was induced using ethyl-methanesulfonate (EMS) to obtain mutants. Among the leaf variegated Hosta plants, some with yellowish-green color pattern on the edge of the leaves was selected. They were cultivated through vegetative production. Assessment of the botanical characteristics was executed for six years from 2013 to 2018. Major characteristics such as yellowish-green color pattern on the edge of leaves appeared uniformly and was finally selected in 2019. The Hosta 'Hwangnarae' that have been bred in this way can prove to be useful as material for a ground cover plant or pot plant.

• Environmental Mutagens and Carcinogens
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• v.18 no.2
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• pp.116-122
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• 1998

The present investigation has been performed to elucidate the effect of pretreatment with low dose of ultraviolet radiation (UV), ethyl methansulfonate (EMS), and bleomycin (BLM) on cell survival and lectin-binding glycoconjugates of plasma membrane in HeLa cells treated with mutagen. The percentage of survival of cells pretreated with 1 mM EMS following treatment with 10 mM EMS was higher than that of cells treated with 10 mM EMS alone. Wheat germ agglutinin (WGA) staining intensity of cells pretreated with 1 mM EMS and subsequently treated with 10 mM EMS was stronger than that of cells treated with 10 mM EMS alone. But, succinylated wheat germ agglutinin (sWGA) staining intensity of cells pretreated with 1 mM EMS and subsequently treated with 10 mM EMS was similar to that of cells treated with 10 mM EMS alone. These results suggest that the acquired resistance to EMS is related to the glycoconjugates containing sialic acid of plasma membrane involved in multidrug resistance or adaptive response in HeLa cells.

• Proceedings of the Zoological Society Korea Conference
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• 1998.10b
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• pp.113-113
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• 1998

In Saccharomyces cerevisiae UV irradiation and a variety of chemical DNA -damaging agents induce the transcription of specific genes, including several involved in DNA repair. One of the best characterized of DNA -damage inducible genes is PHRI, which encodes the apoenzyme for DNA photolyase. Basal-level and damage-induced expression of PHRI require an upstream activation sequence, UASPHRI. Here we report the identification of the UlvIE6 gene of S. cerevisiae as a regulator of UASPHRl activity. Surprisingly, the effect of deletion of UME6 is growth phase dependent. In wild-type cells PHRI is induced in late exponential phase, concomitant with the initiation of glycogen accumulation that precedes the diauxic shift. Deletion of UNIE6 abolishes this induction, decreases the steady-state concentration of photolyase molecules and PHRI mRNA, and increases the UV sensitivity of a rad2 mutant. The results suggest that UM E6 contributes to the regulated expression of a subset of damage-responsive genes in yeast. Furthermore, the upstream repression sequence, URSPHRI, is required for repression and damage-induced expression of PHRl. Here we show identification of YER169W and YDR096W as putative regulators acting through $URS_{PHRI}$. These open reading frames were designated as RPHI (YERl69W) and RPH2 (YDR096W) indicating regulator of PHRI. Simultaneous disruption of both genes showed a synergistic effect, producing a four-fold increase in basal level expression and a similar decrease m the induction ratio following treatment of methyl methanesulfonate(MMS). Mutation of the sequence ($AG_4$) bound by Rphlp rendered the promoter of PHRI insensitive to changes in RPHI or RPH2 status. The data suggest that RPHI and RPH2 act as damage-responsive negative regulators of PHRI. Surprisingly, the sequence bound by Rphlp in vitro is found to be $AG_4$ which is identical to the consensus binding site for the regulators Msn2p and Msn4p involved in stress-induced expression. Deletion of MSN2 and MSN4 has little effect on the induction$.$ ratio following DNA damage. However, all deletions led to a significant decrease in basal-level and induced expression of PHRI. These results imply that MSN2 and MSN4 are positive regulators of P HRI but are not required for DNA damage repression. [Supported by grant from NIH]om NIH]