Annals of Occupational and Environmental Medicine

Original Article

Effects of Selenium on Apoptosis Induced by Methyl Mercury Chloride in RAW 264.7 Cells: Keun Snag Kwon, Dai Ha Koh, Jung Ho Youm, Wook Hee Yoon; Korean Journal of Occupational and Environmental Medicine 2003;15(3):237-251. Published online September 30, 2003; DOI: https://doi.org/10.35371/kjoem.2003.15.3.237

Abstract PDF: OBJECTIVE: This study was performed to evaluate the protective effects of selenium against the methyl mercury chloride (MeHgCl) induced cell apoptosis.
METHODS
The effect of selenium on the MeHgCl induced cell apoptosis was observed in mouse macrophage-derived RAW 264.7 cells, in vitro. The cells were cultured in Dulbecco's modified Eagle's medium (DMEM).
RESULTS
MeHgCl exerted a dose dependent cytotoxicity, as demonstrated by the MTT assay, an assay dependent, in part, on mitochondrial function. Concurrent exposure to selenium provided complete protective effects against the cytotoxicity induced by MeHgCl. Pretreatment with selenium increased the protective effects of subsquent administrations of selenium in conjunction with MeHgCl, but pretreatment of selenium alone did not provide protection against MeHgCl when given alone. Selenium administered after exposure to MeHgCl did not repair the existing MeHgCl induced cytotoxicity.Furthermore, the apoptosis induced by MeHgCl was revealed by the DNA fragmentation, using the terminal deoxynucleotidyl transferase Biotin-dUTP nick end labeling (TUNEL) assay, alterations to the nuclear morphology, by nuclei staining, and the plasma membrane lipid organization, as shown by cell flow cytometry. The apoptosis induced by MeHgCl was prevented by the concurrent exposure to selenium, or pretreatment with selenium, prior to the administration of selenium in conjunction with MeHgCl. However, no inhibittion of the MeHgCl induced apoptosis was observed with selenium pretreatment prior to exposure to MeHgCl alone, or with the administration of selenium after exposure to MeHgCl.
CONCLUSIONS
These results suggest that the coexistence of selenium and MeHgCl are essential for the protective effects of selenium against the MeHgCl-induced apoptosis, and the cytotoxicity, in RAW 264.7 cells, and may involve selenium-MeHgCl binding.

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Original Article

Effect of Glutathione on Apoptosis Induced by Methyl Mercury Chloride: Jung Ho Youm, Dai Ha Koh, Keun Sang Kwon, Me Yae Lee; Korean Journal of Occupational and Environmental Medicine 2002;14(4):377-391. Published online December 31, 2002; DOI: https://doi.org/10.35371/kjoem.2002.14.4.377

Abstract PDF: OBJECTIVES
This study was performed to evaluate the critical role of glutathione(GSH) in methyl mercury chloride(MeHgCl)induced cell apoptosis.
METHODS
The effect of GSH in MeHgCl induced cell apoptosis was observed in mouse macrophage-derived RAW 264.7 cells in vitro. The cells were cultured in Dulbecco's modified Eagle's medium(DMEM).
RESULTS
MeHgCl exerted a dose dependent cytotoxicity,as demonstrated by the MTT assay, which is an assay dependent partially on the mitochondrial function. Moreover, in the presence of NAC, a GSH precursor, the MeHgCl induced cytotoxicity was significantly decreased whereas BSO, a specific GSH synthesis inhibitor,increased the MeHgCl induced cytotoxicity.The MeHgCl induced DNA fragmentation and chromatin condensation was consistent with the morphological alterations. The MeHgCl treated cells exhibited increasing annexin V-FITC binding to the phos-phatidylserine(PS)translocated from the inner to the outer leaflet of the plasma membrane and those cells with NAC pretreatment significantly exhibited decreasing annexin V-FITC binding compared to the cells treated with MeHgCl only. However BSO pretreatment markedly exhibited the increasing annexin V-FITC binding. The MeHgCl treated cells generated ROS, which was evidenced by the oxidation of dihydroethidine and the generation of the fluorescent product, ethidium. In addition, BSO pretreatment further enhanced the extent of ROS generation caused by MeHgCl whereas NAC pretreatment decreased the amount of ROS generation. MeHgCl led to a dose dependent decrease in the GSH content. Although MeHgCl exposure significantly reduced the GSH level, those cells that had a NAC pretreatment contained a higher level of GSH compared to the cells treated with MeHgCl only. In contrast, BSO pretreatment futher enhanced the extent of GSH depletion caused by MeHgCl.
CONCLUSIONS
These results indicate that MeHgCl reduced the GSH content and impaired the defense against oxidative damage caused by ROS formation in RAW 264.7 cells. It is possible that these factors leads to the activation of the apoptosis signaling pathway. Ultimately these results suggest that GSH plays a crucial role in protecting the activity against MeHgCl induced apoptosis.

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Original Article

Cadmium-induced Apoptosis in HL-60 Cells Via Signal Transduction: Nam Song Kim, Gyung Jae Oh, Kwang Ho Cho, Mee Sun Hyun, Yoo Chang Kim, Tae Ho Sung, Jung Ho Youm, Keun Sang Kwon; Korean Journal of Occupational and Environmental Medicine 2002;14(1):1-12. Published online March 31, 2002; DOI: https://doi.org/10.35371/kjoem.2002.14.1.1

Abstract PDF

OBJECTIVES
Apoptosis is a process of active cell death, distinct from necrosis and characterized by specific morphological and biochemical features. Apoptosis induced by metals and metal-related deleterious conditions has only recently been studied. Although the toxic effects of heavy metals are well described, little is known about the mechanism of apoptosis via cadmium toxicity. Therefore, this study is designed to define the induction mechanism of apoptosis by which cadmium exerts its cytotoxic effect on human promyelocytic leukemic HL-60 cells. The cytotoxic effects of cadmium on HL-60 cells are studied in regards to apoptotic signal transduction pathways.
METHODS
The mode of cadmium-induced apoptosis was investigated in HL-60 cells. HL-60 cells were treated with various concentrations of cadmium and antioxidants after which the viability of the cells were measured by MTT assay. The morphological features of cadmium- induced apoptosis were evaluated by fluoromicroscopy and the DNA fragmentation was analyzed using 1.5% agarose gel electrophorosis. Kinase activity was assayed by autoradiography and activity of NF-kappaB and nuclear proteins were measured by EMSA.
RESULTS
Cadmium (125 microM) induces the characteristic morphological features of apoptosis, which are characterized by a shrinkage of the cytoplasm and a condensation of chromatin. In addition, cadmium induced the ladder pattern of DNA fragmentation. Antioxidants(Sodium nitroprusside, glutathione and N-acethylcysteine), which were not toxic to the cells, did not suppress apoptosis induced by cadmium. Cadmium enhances the expression of several classes of genes at elevated cytotoxic concentrations. Poly(ADP-ribose) polymerase(PARP) was predominantly in the fragmented form when doses of 125 microM were used. Since PARP is cleaved by CPP32 (caspase-3), we next determined if cadmium was capable of effecting changes in CPP32 activity. The results of these experiments showed that cadmium increased caspase-3 activity in a time dependent manner, corresponding to the time of appearance of fragmented PARP. Cadmium also increased the phosphotransferase activities of c-JUN N-terminal kinase (JNK). Furthermore, cadmium increased the activation of transcriptional factors including the activation of protein-1 (AP-1) and NF-kappaB .
CONCLUSIONS
These results suggest that cadmium induces the apoptotic death of HL-60 cells via the activation of a DEVD-specific caspase, JNK and transcriptional factors such as AP-1 and NF-kappaB .

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The association between blood cadmium level, frequency and amount of gejang (marinated crab) intake
Chang Yul Choi, Gun Il Park, Young Seok Byun, Man Joong Jeon, Kwang Hae Choi, Joon Sakong
Annals of Occupational and Environmental Medicine.2016;[Epub] CrossRef

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Original Article

Apoptosis Induced by Manganese in Basal Ganglia Primary Neuronal Cell Culture: Morphological Findings: Dong Hoon Shin, Sang Pyo Kim, Young Wook Jung, Jae Hoon Bae, Dae Kyu Song, Won Ki Baek; Korean Journal of Occupational and Environmental Medicine 2000;12(1):41-47. Published online March 31, 2000; DOI: https://doi.org/10.35371/kjoem.2000.12.1.41

Abstract PDF: OBJECTIVES
Manganese is cytotoxic to the central nervous system including basal ganglia. Its toxic mechanism is related to oxidative stress, mediated by toxic free radicals but is specultives. In the present study, we have investigated to manifest apoptosis in manganese-induced cytotoxicity in primary neuronal cell culture of rat basal ganglia.
METHOD
To detect apoptotic neuronal cells were stained by the terminal deoxynu-cleotide(TdT)-mediated dUTP nick end-labelling(TUNEL) method and apoptotic changes in nuclei of neurons were observed by electron microscopy.
RESULTS
We showed that TUNEL immunostain showed brownish signal in the nuclei of apoptotic cells and the proportions of apoptotic cells in Manganese treatment groups were more higher than controls. On transmission electron microscopy, there were chromatine condensation with margination toward nuclear membrane and condensation of cytoplasm in the treated with luM MnC1, for 48 hours in a basal ganglia neurons. Apoptotic bodies were found and consisted of semilunar-like condensed nuclei with relatively intact cytoplasmic organelles.
CONCLUSIONS
Apoptosis appears to be one mechanism in the manganese-induced neuronal cell death. Manganese intoxication is a convenient model for apoptosis study.

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Original Article

Induction of Apoptosis by Heavy Metals in HL-60 Cells: Nam Song Kim, Tae Ho Seong, Kwang Ho Cho, Jung Ho Youm, Dai Ha Koh; Korean Journal of Occupational and Environmental Medicine 1999;11(4):557-568. Published online December 31, 1999; DOI: https://doi.org/10.35371/kjoem.1999.11.4.557

Abstract PDF: OBJECTIVES
Apotosis induced by metals and metal-related deleterious conditions has only recently been studied. Although the toxic effects of heavy metal are well described, little is known about the mechanism of apoptosis by heavy metal toxicity. This study is designed to define the induction of apoptosis by which heavy metals exert the cytotoxic effect on human promyelocytic leukemic HL-60 cells.
Methods
After the incubation with CdC12, Na2SeO3 and HgC12, viability of the cells were measured by MTT assay. DNA fragmentation was analyzed by electrophoresis. For measurement of caspase 1 and 3-like proteases activity, the whole lysates were subjected to the proteolytic cleavage and then measured by using fluorospectrometry. c-JUN N-terminal kinase (JNK) activity was detected by an in vitro kinase assay. Transcriptional activities of activating protein-1 (AP-1) and nuclear factor-kB (NF-kB) were measured by elec trophoresis mobility shift assay (EMSA).
RESULTS
Cadmium (l2OuiN/I) and selenium (30,iM) induce the apoptosis of HL-60 cells which is characterized by the ladder pattern of DNA fragmentation. Cadmium and selenium induce the activation of caspase-3 in a time dependent manner. They also increase the phosphotransferase activities of c-JUN N-terminal kinase (JNK) in cadmium and selenium treated HL-60 cells. Furthermore, cadmium and selenium increase the activation of transcriptional factors including AP-i and NF-kB.
CONCLUSIONS
These results suggest that cadmium and selenium induce the apoptotic death of HL-60 cells via activation of DEVD-specific caspase, JNK and transcriptional factors such as AP-1 and NF-kB.

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Original Article

Apoptosis of Neuronal Cells Induced by Lead: Seon Hee Yang, Dong Hoon Shin, Won Ki Baek; Korean Journal of Occupational and Environmental Medicine 1999;11(2):254-263. Published online June 30, 1999; DOI: https://doi.org/10.35371/kjoem.1999.11.2.254

Abstract PDF

Lead is a major environmental and occupational neurotoxicant. It has been shown that long-term exposure to a low level of lead impairs the development of brain. For example, it was reported that lead exposure during the childhood causes a learning difficulty and a memory deficit of children. Neurotoxic agents including the lead are believed to cause neuronal death in developing brain by two mechanisms: apoptosis and necrosis. However, the exact mechanism of neuronal death caused by lead exposure is still not known explicitly. In this study, we conducted a study to clarify a mechanism of hippocampal neuronal cell death caused by lead acetate. Hippocampal neurons were cultured for 14-16 days and treated with lead acetate of 1. 10, 100 1 microM concentrations for 12 hours. With the MTT(methyl tetrazolium test) kit, the viability of neuronal cells was measured. Next, in order to examine apoptosis caused by lead acetate, TUNEL (TdT-mediated d-UTP Nick End Labelling) assay was performed. It has been shown that lead acetate reduced the viability of neuronal cells in a dose dependent manner, especially at the concentration of 100 ~M lead acetate. TUNEL immunostain showed brownish signals in the nucleus of apoptotic cells. The proportions of apoptotic cells in the lead?acetate treated group were more higher than those in the controls and increased as lead acetate concentration increased. From above results, it may be concluded that lead in the hippocampal neuronal cells reduced cell viability and one of mechanisms in neuronal cell death by lead appears to be apoptosis.

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Teratogenic Effects of Lead in the Developing Chick Embryos
Aqsa Shafiq, . Mahnoor, Mirza Fahad Baig, Yashal Fatima, Arooj Haer, Shakila Parveen, Muhammad Akram Tariq, Muhammad Khalil Ahmad Khan
MARKHOR (The Journal of Zoology).2023; : 16. CrossRef

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Original Article

Silica Induced Apoptosis In Vitro and In Vivo: Ji Hong Kim, Hwang Shn Chang, Yung Mann Baag, Byung Yong Ahn, Kyoung Ah Kim, Young Lim; Korean Journal of Occupational and Environmental Medicine 1999;11(2):206-212. Published online June 30, 1999; DOI: https://doi.org/10.35371/kjoem.1999.11.2.206

Abstract PDF: Silica exposure results in acute inflammatory response followed by chronic fibrotic change. The mechanism for the maintenance of silica-induced inflammation has not been understood yet. Apoptosis is a morphologically distinct form of programed cell death that plays major role during homeostasis and in many diseases including cancer, acquired immune deficiency syndrome and neurodegenerative disorders. Apoptosis is characterized by cell shrinkage, membrane blebbing and nuclear condensation. To demonstrate the involvement of apoptosis in underlying mechanism for the development of silica-induced pathological changes, this study was designed in vitro and in vivo models. In in vitro study, alveolar epithelial cell line (A549) was stimulated with silica and performed flow cytometry and DNA electrophoresis. In in vivo study, bronchoalveolar lavage (BAL) was done to count the total and apoptotic cells from silica-instilled rats. The results were as follows: 1. Apoptotic cell fraction of silica-treated groups (10 and 50 microgram/cm2) was significantly higher than that of control group. 2. Genomic DNA from silica-treated groups (10 and 50 microgram/cm2 ) showed DNA ladder in agarose gel electrophoresis, while group of 1 microgram/cm2 didn't. 3. Total cell number and apoptotic cell number of BAL fluid from silica-instilled rats (10, 20 and 40 mg/kg) were significantly higher than those of control. 4. Silica induced apoptosis of cells in BAL fluid was confirmed by microscopic observation with nuclear fragmentation. These results suggest that apoptosis may contribute to development of silica-induced pathological changes.

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Original Article

Cytotoxicity and Apoptosis by Silica, Asbestos and Man-Made Mineral Fibers: Young Lim, Kyoung Ah Kim, Heung Nam Kim, Dong Won Lee, Won Seop Cho, Im Goung Yun; Korean Journal of Occupational and Environmental Medicine 1997;9(4):641-649. Published online December 31, 1997; DOI: https://doi.org/10.35371/kjoem.1997.9.4.641

Abstract PDF

Exposure to various particles and fibers can result in lung inflammation that may progress to fibrosis, even lung cancer for which there is no effective clinical treatment now. The mechanism involved in pulmonary injury has not been well defined ; however, most current evidence implicates a central role for alveolar macrophages (AM) in this process. Also apoptosis or programmed cell death is regarded as a mechanism which is related with the pulmonary fibrosis. We propose that the cytotoxic potential of various particles may be evaluated by measuring lactic dehydrogenase (LDH) from particle co-cultured supernatant and theses particles may induce the characteristics of apoptosis, DNA ladder. We analyzed rat AM culture media which was incubated for 3 days with the same concentration (10 ug/ml) of silica(Si), chrysotile(Ch), crocidolite(Cr), ceramic fiber(CF), rock wool(RW) and glass wool (GW). And each particles (50ug/cm(2)) was incubated with A549 (pneumocyte in tracheal epithelium) cell lines for 24 hours to confirm the DNA ladder. Additionally, silica induced apoptosis in vivo was confirmed by electromicroscopic observation. The results were as follows; 1. Silica, asbestos and man-made mineral fibers (MMMF) co-cultured with AM showed the increase of LDH significantly with the time interval of 24, 48, 72 hours except for ceramic fiber in 48 and 72 hours and crocidolite in 72 hours. 2. Silica, asbestos and man-made mineral fibers (CF, GF) showed the characteristics of apoptosis, DNA ladder, which was induced by incubating A549 cell with each particles for 24 hours in vitro 3. Apoptotic alveolar macrophage was observed the findings of zeiosis (membrane blebbing), condensation of nuclear chromosome and many vacuoles in cytoplasm, electomicroscopically.

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Occupational Cancers with Chemical Exposure and their Prevention in Korea: A Literature Review
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Asian Pacific Journal of Cancer Prevention.2013; 14(6): 3379. CrossRef
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Tian Zhu Li, Soo-Jin Lee, Se-Jong Park, Byung-Joon Chang, Jong-Hwan Lee, Kil-Soo Kim, Myoung-Heon Lee, Nong-Hoon Choe
Tuberculosis and Respiratory Diseases.2006; 60(5): 554. CrossRef

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