• Title/Summary/Keyword: DNA Fragmentation

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Apoptotic Cell Death of Mouse Splenocytes by Polychlorinated Biphenyls and Its Prevention by Serum

  • Yoo, Byung-Sun;Kim, Hwan-Mook
    • Toxicological Research
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    • v.13 no.3
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    • pp.187-191
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    • 1997
  • Cell death induced by polychlorinated biphenyls (PCBs), environmental toxicant, was investigated in mouse splenocytes. The fragmentation of intact DNA, a parameter of apoptotic cell death, was evaluated qualitatively by agarose gel electrophoresis analysis and quantitatively by diphenylamine reaction method. PCBs induced apoptotic cell death of splenocytes in a dose- and time- dependent manner. The effect of serum on the apoptotic cell death induced by PCBs was also investigated. The DNA fragmentation induced by PCB treatment in serum-free medium was clearly inhibited by an addition of serum to the culture medium. The decrease of DNA fragmentation due to serum addition was accompanied with the increase of cell viability.

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Optimazation of the Assement and Apotosis of Endocrine-Bisphenol A Disruptors (내분비계장애물질 평가방법의 최적화 및 Apoptosis에 관한 연구)

  • Ahn Kwang-Hyun;Lee Kyung-A;Kim Bong-Hee
    • Environmental Analysis Health and Toxicology
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    • v.19 no.3
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    • pp.251-259
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    • 2004
  • Xenoestrogens are chemicals with diverse structure that mimic estrogen. Bisphenol A, a monomer of polycarbonate and epoxy resins, has been detected in canned food and human saliva. Bisphenol A stimulate cell proliferation and induce expression of estrogen -response genes in vitro. The purpose of the this study was to evaluate cell proliferation of bisphenol A in the presence of a rat liver 59 mix contaning cytochrome P450 enzymes and Cu (II). The fragmentation of intact DNA, a parameter of apoptotic cell death, was evaluated quantitatively by diphenylamine reaction method. Bisphenol A induced apoptotic cell death in a dose-dependent manner The effect of radical scavenger on the apoptotic cell death induced bisphenol A was investigated. The DNA fragmentation induced by bisphenol A was significantly inhibited by addition of radical scavenger to the culture medium. This indicated that elevated oxidative stress caused by imbalance between the production and removal of free radicals occurred in cells. Taken together, these results suggest that free radical reacts with Cu (II) leading oxidative stress.

Effect of AC-264, a Novel Indole Derivative, on Apoptosis in HL-60 Cells

  • Lee, Kyeong;Kwon, Ok-Kyoung;Xia, Yan;Ahn, Kyung-Seop
    • Bulletin of the Korean Chemical Society
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    • v.31 no.12
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    • pp.3777-3781
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    • 2010
  • The anticancer effect and apoptotic mechanism of a novel indole derivative AC-264, a lead derived from a chemical library, were investigated in human promyelocytic leukemia HL-60 cells. HL-60 cells treated with AC-264 at various concentrations showed the morphological features of apoptosis, such as plasma membrane blebbing and cell shrinkage. AC-264 exhibited cytotoxic effect in various cancer cell lines with different degrees of potency. Especially, AC-264 was effective on increasing the population of apoptotic cells in HL-60 cells, as detected by the number of cells stained with Annexin V and PI. Furthermore, AC-264 activated caspase-3 enzyme activity and induced internucleosomal DNA fragmentation. These results indicated that AC-264 produces anti-cancer effect via apoptotic cell death by activating caspase-3 and inducing internucleosomal DNA fragmentation in HL-60 cells.

Assessements of Apoptosis in Bovine Embryos Reconstructed with Fetal Fibroblast

  • Lee, S. L.;Park, G.;S. Y. Choe
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.136-136
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    • 2003
  • Mainly due to deficiencies in nuclear reprogramming, gene expression and DNA fragmentation, which result in early and late embryonic losses, the overall success rate achieved by cloning techniques to date is low. This present study compared the incidences of DNA fragmentation during development of IVF, parthenotes (PT), nuclear transfer (NT) and transgenic (TG) embryos. Terminal deoxynucleotidyl transferase (TdT) nick-end labelling (TUNEL) with propidium iodide counter staining was used for determination of DNA fragmentation and total number, respectively. TG and NT donor cells were fetal fibroblasts with or without transfection with EGFP, and cultured in DMEM+15% FCS until confluent, for 5 days. At 19 h post-maturation (hpm), enucleated oocytes were reconstructed with donor cells and activated at 24 hpm with the combinations of ionomycin (5 M, 5 min) and cyclo-heximide (10 g/ml, 5 h) after electric fusion by a single DC pulse (1.6 KV/cm, 60 sec). Parthenotes were produced by the same activation protocol at 24 hpm. (중략)

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Circulating DNA in Egyptian Women with Breast Cancer

  • Ibrahim, Iman Hassan;Kamel, Mahmoud M;Ghareeb, Mohamed
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.6
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    • pp.2989-2993
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    • 2016
  • The commonest cancer in Egyptian females occurs in the breast cfDNA is a non-invasive marker for tumor detetion and prognostic assessment in many types of cancer including breast cancer. This study aimed to assess the role of cfDNA and its fragmentation pattern in breast cancer prognosis and treatment response. Forty female patients with malignant breast tumors and a comparable group of healthy blood donors were enrolled prospectively. cfDNA levels and fragmentation patterns were investigated after cfDNA extraction, gel electrophoresis and gel analysis. The percentage of breast cancer patients positive for cfDNA (92.5%) was significantly higher than that of controls (55%). Also, mean concentration of cfDNA was significantly higher than in the control group (P<0.05). Most Her-2 positive patients had long cfDNA fragments, this being significant as compared to Her-2 negative patients (P<0.05). Metastasis was also positively linked to significantly higher cfDNA (P<0.05) and the mean cfDNA integrity index was significantly higher in non-responders compared to treatment responders (P<0.05). In conclusion, both qualitative and quantitative aspects of cfDNA and its different fragments in breast cancer patients could be related to prognosis, metastasis and treatment response. Long cfDNA fragments could be particularly useful for prediction purposes.

Effects of Yuldahansotang after kainate administration in the mouse hippocampus area (열다한소탕(熱多寒少湯)이 kainic acid에 의해 유발된 mouse의 해마체 손상에 미치는 영향)

  • Kim, Il-hwan;Kim, Kyung-yo
    • Journal of Sasang Constitutional Medicine
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    • v.11 no.2
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    • pp.283-299
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    • 1999
  • 1. Purpose : Systemic injection of kainic acid in experimental animals induces the limbic seizure and structural damages in hippocampus and amygdala which resembles the changes in human temporal lobe epilepsy. The author performed this study to investigate the neuroprotective effects of Yuldahansotang, on the neurotoxicity induced by kainic acid in the hippocampus in rats. 2. Method : Kainic acid was administered intraperitoneally. And feeding with Yuldahansotang for 3 weeks after kainic acid administration. Seizure were induced in male mice (kainate 10-40 mg/kg i.p) and animals were sacrified at various time-points after injection. The experimental animals were sacrificed at 1, 2, 3day and 1, 3weeks while Yuldahansotang administrations. Seizure were graded using a behavioral scale developed in our laboratory. c-fos belong to immediate early genes(IEGs) known to have rapid and brief responses. And neuronal injury was assayed by examining DNA fragmentation using in situ nick translation histochemistry. 3. Results & Conclusion : Seizure severity paralled kainate dosage. At higher doses DNA fragmentation is seen mainly in hippocampus in area CA3, and variable in CA1, thalamus, amygdala within 24 h, is maximal within 72 h, and is large gene by 7 days after administration of kainate. And we can't see the expression of DNA fragmentation and c-fos in the mice what feeded by Yuldahansotang after 7 days from kainic acid administration. It is consequently suggested that Yuldahansotang may attenuate the kainic acid-induced neuronal degeneration and increase the immunoreactivity of hippocampus in mouse.

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Inducible Nitric Oxide Synthase Expression and Luteal Cell DNA Fragmentation of Porcine Cyclic Corpora Lutea

  • Tao, Yong;Fu, Zhuo;Xia, Guoliang;Lei, Lei;Chen, Xiufen;Yang, Jie
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.626-631
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    • 2005
  • Nitric oxide (NO) derived from inducible nitric oxide synthase (iNOS) is involved in cell apoptosis, which contributes to luteal regression and luteolysis in some species. In large domestic animals, no direct evidence for the relationship between NO and cell apoptosis in the process of corpus luteum regression is reported. The present study was conducted to investigate the localization of iNOS on porcine corpora lutea (CL) during the oestrus cycle and its relation to cell DNA fragmentation and CL regression. According to morphology, four luteal phases throughout the estrous cycle were defined as CL1, CL2, CL3 and CL4. By isoform-specific antibody against iNOS, the immunochemial staining was determined. Luteal cell DNA fragmentation was determined by flow cytometry. The results showed that no positive staining for iNOS was in CL1 and that iNOS was produced but limited to the periphery of CL2, while in the CL3, the spreading immunochemical staining was found inside the CL. No iNOS positive staining was detected in CL4. Meanwhile, DNA fragmentation increased dramatically when CL developed from CL2 to CL3 (p<0.05). In CL4, higher proportion of luteal cells still had fragmented DNA than that of luteal cells from CL1 or CL2 (p<0.05). These results indicate that iNOS expression is closely related to luteal cell apoptosis and then to luteal regression.

In vitro Investigation of Cytotoxic and Apoptotic Effects of Cynara L. Species in Colorectal Cancer Cells

  • Simsek, Ela Nur;Uysal, Tuna
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6791-6795
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    • 2013
  • Apoptotic and cytotoxic activity of plant extracts obtaining from naturally growing Cynara syriaca in Turkey and cultivated C cardunculus against DLD1 colorectal cancer cells was determined. Extracts from wild and cultivated Cynara species were obtained from their vegetative parts and receptacles using hexane and applied with five different dose (0.1-1 mg/ml) as well as apigenin for MTT tests for three time periods (24, 48 and 72 hours). After cells were treated with $IC_{50}$ doses for each extract total DNA and RNA were isolated for determination of the cause of cell death. From isolated RNAs, cDNA were synthesized and amplification of p21, BCL-2 and BAX gene regions was carried out. Consequently, we found that pro-apoptotic (BAX) gene expression and a cell cycle inhibitor (p21) were induced in the presence of our artichoke extracts. In contrast, anti-apoptotic BCL-2 gene expression was reduced compared to the control group. In addition DNA fragmentation results demonstrated DLD1 cell death via apoptosis.

Fungal Taxol Extracted from Cladosporium oxysporum Induces Apoptosis in T47D Human Breast Cancer Cell Line

  • Raj, Kathamuthu Gokul;Sambantham, Shanmugam;Manikanadan, Ramar;Arulvasu, Chinnansamy;Pandi, Mohan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6627-6632
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    • 2014
  • Purpose: The present study concerns molecular mechanisms involved in induction of apoptosis by a fungal taxol extracted from the fungus Cladosporium oxysporum in T47D human breast cancer cells. Materials and Methods: Apoptosis-induced by the fungal taxol was assessed by MTT assay, nuclear staining, DNA fragmentation, flow cytometry and pro- as well as anti-apoptotic protein expression by Western blotting. Results: Our results showed inhibition of T47D cell proliferation with an $IC_{50}$ value of $2.5{\mu}M/ml$ after 24 h incubation. It was suggested that the extract may exert its anti-proliferative effect on human breast cancer cell line by suppressing growth, arresting through the cell cycle, increase in DNA fragmentation as well as down-regulation of the expression of NF-${\kappa}B$, Bcl-2 and Bcl-XL and up-regulation of pro-apoptotic proteins like Bax, cyt-C and caspase-3. Conclusions: We propose that the fungal taxol contributes to growth inhibition in the human breast cancer cell through apoptosis induction via a mitochondrial mediated pathway, with possible potential as an anticancer therapeutic agent.