• Title/Summary/Keyword: Hamster

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Modulating Effect of Lupeol on the Expression Pattern of Apoptotic Markers in 7, 12-Dimethylbenz(a)anthracene Induced Oral Carcinogenesis

  • Manoharan, S.;Palanimuthu, D.;Baskaran, N.;Silvan, S.
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5753-5757
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    • 2012
  • Apoptosis, also known as cell suicide or programmed cell death, removes unwanted and genetically damaged cells from the body. Evasion of apoptosis is one of the major characteristic features of rapidly proliferating tumor cells. Chemopreventive agents inhibit or suppress tumor formation through apoptotic induction in target tissues. The aim of the present study was to investigate the pro-apoptotic potential of lupeol during 7,12-dimethylbenz(a) anthracene (DMBA) induced hamster buccal pouch carcinogenesis. Topical application of 0.5% DMBA three times a week for 14 weeks in the buccal pouches of golden Syrian hamsters resulted in oral squamous cell carcinoma. The expression pattern of apoptotic markers was analyzed using immunohistochemistry (p53, Bcl-2, Bax) and ELISA reader (caspase 3 and 9). In the present study, 100% tumor formation with defects in apoptotic markerexpression pattern was noticed in hamsters treated with DMBA alone. Oral administration of lupeol at a dose of 50mg/kg bw completely prevented the formation oral tumors as well as decreased the expression p53 and Bcl-2, while increasing the expression of Bax and the activities of caspase 3 and 9. The present study thus indicated that lupeol might inhibit DMBA-induced oral tumor formation through its pro-apoptotic potential in golden Syrian hamsters.

Expression and Purification of Recombinant Human Angiopoictin-2 and Its Analog in Chinese Hamster Ovary Cells

  • Hwang, Su-Jeong;Kim, In-Jun;Go, Gyu-Yeong;Lee, Gyun-Min
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.459-462
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    • 2001
  • Angiopoietin-2 (Ang2) is a naturally occurring antagonist for angiopoietin-l (Angl) and its Tie2 receptor during vasculogenesis, Although angiopoietins have been expressed in several mammalian cell lines, their expression levels are low. Recombinant Chinese hamster ovary (CHO) cell lines expressing a high level of human Ang2 or its analog, human $Ang2_{443}$, with an amino-terminal FLAG-tag were constructed by transfecting the expression vectors into dhfr-deficient CHO cells and subsequent gene amplification in medium containing stepwise increments in methotrexate level. Secreted Ang2 or human $Ang2_{443}$ were purified from the cultured medium using an anti-FLAG- agarose affinity chromatography, The purified Ang2 and $Ang2_{443}$ migrated on SOS-PAGE as a broad band, characteristic of glycosylated protein. Their biological activity in vitro was demonstrated in a serum deprivation-induced apoptosis assay. Ang2 at high concentration, like AngI, can be an apoptosis survival factor for endothelial cells through the activation of the Tie2 receptor.

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Effect of Anti-apoptotic Agents in Recombinant Chinese Hamster Ovary Cell Expressing Iduronate 2-sulphatase

  • Jeon, Ok-Seon;Chun, Ju-Me;Kang, Seon-Ah;Lee, Sang-Jong;Chun, Gie-Taek;Chang, Yong-Keun;Jeong, Yeon-Ho
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.210-213
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    • 2005
  • The suppression of apoptosis during the cell culture might increase recombinant protein production. In the present study, the effects of anti-apoptotic agents on the apoptosis of recombinant Chinese Hamster Ovary cells and the production of Iduronate 2-sulphatase(IDS) were investigated Cell density slightly increased when $2{\mu}M$ of EGCG and $10{\mu}g/mL$ of STR-G were added to culture medium after two days. It was observed that the percentage of apoptotic cells was decreased in the culture with STR-G, and Bcl-2 expression level was enhanced in both culture with STR-G and EGCG. These results suggest that G418 and EGCG are effective anti-apoptotic agents for increasing the productivity of IDS with recombinant CHO-DG44.

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Adaptive Response in Chinese Hamster lung Cells by Benzidine Dihydrochloride (Benzidine dihydrochloride에 의한 Chinese hamster lung 세포의 적응반응)

  • 맹승희;정해원;이권섭;이용묵;정호근;유일재
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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    • pp.142-148
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    • 2001
  • We studied adaptive response in CHL cells by benzidine dihydrochloride, a derivative of benzidine, which was a major mutagenic agent in dye industry. Chromosome aberration analysis was used for the identification of adaptive response to this mutagen. Adaptive and reactive doses were confirmed by cell proliferation rate curve. Cell proliferation rate curve was obtained from the mitotic indices of cells treated with various concentrations of benzidine dihydrochloride for 24 hours. Marked adaptive responses to benzidine dihydrochloride in the induction of chromosome aberration were observed in CHL cells by pre-treatment with low concentrations of benzidine dihydrochloride (0.0047 mg/$m\ell$ or 0.0094 mg/$m\ell$) for 24 hours following post-treatment with high concentrations (0.0187, 0.0375, 0.075, 0.15 mg/$m\ell$) for 24 hours. These adaptive responses were found mostly in the type of chromatid breaks and chromatid exchanges. There is no difference in these results between two adaptive doses, 0.0047 mg/$m\ell$ and 0.0094 mg/$m\ell$. The amount of adaptive response, however, was dependent on post-treatment doses.

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Evaluation of the Genetic Toxicity of Synthetic Chemicals (XIII) - Single Cell Gel Electrophoresis of Benzoyl Chloride, 2-Propyn-1-ol, and 2-Phenoxyethanol in Chinese Hamster lung Fibroblast -

  • Ryu, Jae-Chun;Kim, Youn-Jung
    • Environmental Mutagens and Carcinogens
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    • v.24 no.2
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    • pp.79-84
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    • 2004
  • Three synthetic chemicals, benzoyl chloride, 2-propyn-l-ol, and 2-phenoxy ethanol were selected for genotoxicity testing, based on production quantity and available genotoxic data. In our previous report, benzoyl chloride induced chromosomal aberrations in Chinese hamster lung (CHL) fibroblast in vitro with and without metabolic activation, while 2-propyn-l-ol and 2-phenoxy ethanol induced only with metabolic activation. To compare the genotoxicity of chromosome aberration assay, the single cell gel electrophoresis (comet) assay subjected using CHL cells. As a result, statistically significant differences of tail moment values of benzoyl chloride, 2-propyn-1-ol, and 2-phenoxy ethanol were observed compared with control values on almost all concentrations with S9 or without S9 metabolic activation system. This results suggest that genotoxic results of the comet assay and the chromosome aberration assay show correlationship of genotoxicity in the CHL fibroblast. In summary, the positive result of chromosome aberration of benzoyl chloride, 2-propyn-l-ol, and 2-phenoxy ethanol was also induced DNA damages in comet assay with same cell line. Consequently, comet assay will be useful and more accurate tool to detect and to confirm the genotoxicity especially DNA damages in CHL fibroblast.

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Effect of Carcinogenic Chromium(VI) on Cell Death and Cell Cycle in Chinese Hamster Ovary Cells

  • Lee, San-Han;Nam, Hae-Seon;Kim, Sung-Ho
    • Environmental Mutagens and Carcinogens
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    • v.24 no.3
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    • pp.113-120
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    • 2004
  • Chromium compounds are known human and animal carcinogens. In this study, the effects of sodium chromate on apoptosis and cell cycle were investigated in order to unveil the elements of early cellular responses to the metal. Using Chinese hamster ovary cells(CHO-K1-BH4), we found taht chromium (VI) treatment induced apoptosis in these cells, as signified by nuclear fragmentation, DNA laddering on agarose gel electrophoresis, and an increased proportionof cells with hypodiploid DNA. Preceding these changes, chromium (VI) treatment increased caspase 3 pritease activity and also increased expression of p53 protein, while the level of bcl2 protein was not changed. Coincubation with caspase inhibitor, Z-DEVD-FMK, inhibited chromium-induced apoptosis. In the flow cytometric analysis using propidium iodide fluorescence, an increase of cell population in G2/M phase was shown in cells exposed to at least 160 $\mu\textrm{m}$ of sodium chromate for 72h, form 9.8% for 0$\mu\textrm{m}$ chromium (VI) to 26.4% for 320$\mu\textrm{m}$ chromium(VI). Taken together, these findings suggest that chromium(VI)-induced apoptosis is accompanied by G2/M cell cycle arrest, and that p53-mediated pathway may be involved in positive regulation of G2/M arrest and a concurred apoptosis in CHO cells.

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Development of Solid-Gelatin Microcarrier for Large Scale Production of Anchorage-Dependent Animal Gell Lines (부착성 동물세포의 대량배양을 위한 젤라틴 미립담체의 개발)

  • 박정극
    • KSBB Journal
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    • v.4 no.1
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    • pp.18-20
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    • 1989
  • Solid gelatin microcarrier with the size distribution between $100{\mu}{\textrm}{m}$ and $400{\mu}{\textrm}{m}$ was prepared for the attachment and growth experiment for anchorage-dependent animal cell lines, i.e., L 929 and BHK 21. The growth and the maximum cell densities on this gelatin based and polyacrylamide (PAA) microcarriers were compared with those on the commercial dextran based Cytodex 3 microcarrier. Both cell lines showed good comparable attachment and growth patterns on the above three microcarriers. The mouse fibroblast, L 929 showed about the same maximum cell density on PAA, gelatin and Cytodex 3 MC'S BUT BHK 21, the baby hamster kidney cell line, showed the best result on Cytodex 3, which was about $4\times10^6$ cells/ml with the microcarrier concentration of 10 g/1.

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Antimutagenic Effects of Persimmon Leaf tea Extracts in Sister Chromatid Exchanges(SCE) Assay System (감잎차 추출액의 Sister Chromatid Exchange(SCE) 방법에 따른 항돌연변이 효과)

  • 강명희;송현순;이현걸;장해동;김종익;박옥진;이미숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.25 no.2
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    • pp.232-239
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    • 1996
  • 돌연변이 유발 물질인 mitomycin C(MMC)를 처리하여 배양한 Chinese hamster ovary(CHO) cell에 대한 감잎차 추출액의 항돌연변이 효과를 자매 염색 분체 교환(sister chromatid exchange, SCE) 시험법을 사용하여 측정하여 보았다. 감잎차 추출액 자체는 CHO 세포의 SCE 빈도수를 변화시키지 않았으며, 세포의 분열 주기중 S phase에 S9 mixture 없이 감잎차 추출액이 처리되었을 경우 MMC로 유도된 SCE 빈도수를 감소시키지 않았다. 그러나 S9 mixture 존재하에 $G_{1}$ phase에서 MMC 처리 후 감잎차를 처리하는 후처리 방식으로 감잎차 추출액을 처리하였을 때, 저농도($\leq$40$\mu\textrm{g}$/ml)에서 MMC로 인해 유발된 SCE 빈도수가 낮아지는 것을 볼 수 있었다. 이에 비해 고농도(>40$\mu\textrm{g}$/ml)에서는 SCE 빈도수의 감소 효과가 없었다. 본 연구결과, MMC 처리된 CHO 세포에 대한 감잎차 추출액의 항돌연변이 효과를 볼 수 있었고, 이 효과는 S9 mixture 존재하에서 저농도의 감잎차 추출액이 $G_{1}$ phase에 처리되었을 때 나타났다. 감잎차 추출액의 이러한 항돌연변이의 효과의 기전은 감잎차 추출액의 대사산물이 MMC 처리된 CHO 세포에 대한 DNA-excision repair activity를 촉진시키기 때문인 것으로 생각된다.

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Carbohydrate Structure of N- and O-linked Oligosaccharides of Human Erythropoietin Expressed in Chinese Hamster Ovary Cells

  • Lee, Dong-Eok;Ha, Byung-Jhip;Kim, Suk-Joon;Park, Ji-Sook;Yoo, Ree-Ann;Oh, Myung-Suk;Kim, Hyun-Su
    • BMB Reports
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    • v.29 no.3
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    • pp.266-271
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    • 1996
  • A recombinant human erythropoietin (EPO), expressed in Chinese hamster ovary (CHO) cells, is glycosylated at Asn 24, Asn 38, Asn 83, and Ser 126. After release of the N-linked carbohydrate chains by $peptide-N^{4}-(N-acetyl-{\beta}-glucosaminyl)$ asparagine amidase F, the oligosaccharides were analyzed by FACE (Fluorophore-Assisted Carbohydrate Electrophoresis). The O-linked carbohydrate chain was separated by hydrazine, and analyzed by FACE. The monosacccharide composition of recombinant EPO showed man nose, fucose, galactose, N-acetylglucosamine, N-acetylneuraminic acid, and a trace of N-acetylgalactosamine, which are typical monosaccharides in the glycoproteins from the CHO cell. Sequences of N-linked and O-linked oligosaccharides were determined. The structure and composition of oligosaccharides attached to recombinant human EPO, expressed in the CHO cell, are identical to the reported oligosaccharide structure in human EPO isolated from urine.

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Effects of Photoperiod Treatment on Histological Changes in Testis Tissues of the Golden Hamster

  • Kang, Jae-Won;Kim, Seol-Ah;Park, Chang-Eun
    • Korean Journal of Clinical Laboratory Science
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    • v.44 no.1
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    • pp.31-37
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    • 2012
  • Many mammals in temperate zones are affected by the distinctive changes of the four seasons in these zones. Their reproductive status is active in the summer climate and inactive during severe winter weather. The golden hamster (Mesocricetus auratus) is seasonal breeding animal whose sexual activities are regulated by photoperoidism. The reproduction and metabolism are activated by long summer days (LD) and inhibited by short winter days (SD). After several months of SD, animals become refractory to this inhibitory photoperiod and spontaneously revert to LD-like physiology. The suprachiasmatic nuclei (SCN) house the primary circadian oscillator in mammals. Seasonal changes in the photic input to this structure control many annual physiological rhythms via SCN-regulated pineal melatonin secretion, which provides an internal endocrine signal representing photoperiod. The aim of this study was to assess the variation in the morphology of the testis in relation to the natural photoperiod in male golden hamsters. The hamsters were castrated at different weeks (2, 5, 8, and 15). The cell numbers of tubules with spermatogonia (SG), spermatocyte (SC), spermatids (ST), and spermatozoa (SZ) were recorded in each sample. The results showed that testicular regression of golden hamsters occurred in the SD-treated animals. The present investigation determines that the effects of the photoperiod on the reproduction of male golden hamsters. It was also found that the circadian period increases the rate of reproductive inhibition in animals exposed to inhibitory photoperiods.

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