• Title/Summary/Keyword: In vitro irradiation

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Synergistic effect of ionizing radiation and $\beta$-lapachone against tumor in vitro and in vivo

  • Park, Eun-Kyung;Kim, Young-Seok;Lee, Sang-wook;Ahn, Seung-Do;Shin, Seong-Soo;Park, Heon-Joo;Song, Chang-Won
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.80-80
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    • 2003
  • ${\beta}$-lapachone(${\beta}$-Lap), a natural o-naphthoquinone, presents in the bark of the Lapacho tree. ${\beta}$-Lap is cytotoxic against a variety of human cancer cells and it potentiates the anti-tumor effect of Taxol. In addition, ${\beta}$-Lap has been reported to radiosensitize cancer cells by inhibiting the repair of radiation-induced DNA damage.In the present study, we investigated the cytotoxicity of ${\beta}$-Lap against RKO human colorectal cancer cells as well as the combined effect of ${\beta}$-LaP and ionizing radiation. An incubation of RKO cells with 5 ${\mu}$M of ${\beta}$-Lap for 4 h killed almost 90% of the clonogenic cells. An incubation of RKO cells with 5 ${\mu}$M of ${\beta}$-Lap for 4 h or longer also caused massive apoptosis. Unlike other cytotoxic agents, ${\beta}$-Lap did not increase the expression of p53 and p21 and it suppressed the NFkB expression. The expression of Caspase 9 and 3 was minimally altered by ${\beta}$-Lap. Radiation and ${\beta}$-Lap acted synergistically in inducing clonogenic cell death and apoptosis in RKO cells when ${\beta}$-Lap treatment was applied after but not before the radiation exposure of the cells. Interestingly, a 4 h treatment with 5 ${\mu}$M of ${\beta}$-Lap starting 5 h after irradiation was as effective as that starting immediately after irradiation. The mechanisms of ${\beta}$-Lap-induced cell killing is controversial but a recent hypothesis is that ${\beta}$-Lap is activated by NAD(P)H: quinone-onidoreductase (NQO1) in the cells followed by an elevation of cytosolic Ca$\^$2+/ level and activation of proteases leading to apoptosis. It has been reported that NQO1 level in cells is markedly up-regulated for longer than 10 h after irradiation. Indeed, using immunological staining of NQO1, we observed a significant elevation of NQO1 expression in RKO cells 5h after 2-4 Gy irradiation. Such a prolonged elevation of NQO1 level after irradiation may be the reasons why the ${\beta}$-Lap treatment applied S h after irradiation was as effective as that applied immediately after irradiation in killing the cells. In view of the fact that the repair of radiation-induced damage is usually completed within 1-2 h after irradiation, it is highly likely that the ${\beta}$-Lap treahment applied 5 h after irradiation could not inhibit the repair of radiation-induced damage. For in vivo study, RKO cells were injected S.C. into the hind-leg of Nu/Nu mice, and allowed to grow to 130 mm3 tumor. The mice were i.p. injected with ${\beta}$-lapachone or saline 2 h after irradiation of tumors with 10 Gy of X-rays. The radiation induced growth delay was increased by 2.4 $\mu\textrm{g}$/g of ${\beta}$-lapachone. Taken together, we may conclude that the synergistic interaction of radiation and ${\beta}$-Lap in killing cancer cells is not due to radiosensitization by ${\beta}$-Lap but to an enhancement of ${\beta}$-Lap cytotoxicity by radiation through an upregulation of NQO1. The fact that NQO1 is elevated in tumors and that radiation causes prolonged increase of the NQO1 expression may be exploited to preferentially kill tumor cells using ${\beta}$-Lap in combination with radiotherapy.

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Radioprotective effect of naringin and naringenin against cellular damage and oxidative stress of γ-irradiated mice (감마선을 조사한 마우스의 세포 손상과 산화적 스트레스에 대한 나린진과 나린제닌의 방사선방호 효과)

  • Kang, Jung Ae;Kim, Hye Rim;Yoon, Seon Hye;Jang, Beom-Su;Choi, Dae Seong;Park, Sang Hyun
    • Korean Journal of Food Science and Technology
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    • v.49 no.6
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    • pp.659-667
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    • 2017
  • The present study was designed to evaluate the antioxidant activity and radioprotective effects of Naringin and Naringenin in ${\gamma}$-irradiated mice. The antioxidant activity of Naringin and Naringenin was evaluated by 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and ferric reducing antioxidant power (FRAP) assays. Healthy female BALB/c mice were administered Naringin and Naringenin orally ($90{\mu}M/dose$ and $180{\mu}M/dose$) for 7 consecutive days prior to ${\gamma}$-irradiation (6 Gy). Naringenin displayed a much higher antioxidant activity in ABTS and FRAP than naringin. ${\gamma}$-irradiation resulted in cellular damage with decreased spleen and thymus indices and white blood cells (WBC) count. Additionally, ${\gamma}$-irradiation significantly increased lipid peroxidation and decreased the levels of antioxidant enzymes and glutathione (GSH) in the liver tissue. Strikingly, prior administration of Naringenin resulted in considerable prevention of these symptoms. Protection against ${\gamma}$-irradiation-induced cellular damage by Naringenin is likely due to its higher its antioxidant activity. Together, these results confirm that Naringenin is a potent antioxidant and radioprotector.

IGF-1 from Adipose-Derived Mesenchymal Stem Cells Promotes Radioresistance of Breast Cancer Cells

  • Yang, Hui-Ying;Qu, Rong-Mei;Lin, Xiao-Shan;Liu, Tong-Xin;Sun, Quan-Quan;Yang, Chun;Li, Xiao-Hong;Lu, Wei;Hu, Xiao-Fang;Dai, Jing-Xing;Yuan, Lin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10115-10119
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    • 2015
  • Purpose: The aim of this study was to investigate effects of adipose-derived mesenchymal stem cells (AMSCs) on radioresistance of breast cancer cells. Materials and Methods: MTT assays were used to detect any influence of AMSC supernatants on proliferation of breast cancer cells; cell migration assays were used to determine the effect of breast cancer cells on the recruitment of AMSCs; the cell survival fraction post-irradiation was assessed by clonogenic survival assay; ${\gamma}$-H2AX foci number post-irradiation was determined via fluorescence microscopy; and expression of IGF-1R was detected by Western blotting. Results: AMSC supernatants promoted proliferation and radioresistance of breast cancer cells. Breast cancer cells could recruit AMSCs, especially after irradiation. IGF-1 derived from AMSCs might be responsible for the radioresistance of breast cancer cells. Conclusions: Our results suggest that AMSCs in the tumor microenvironment may affect the outcome of radiotherapy for breast cancer in vitro.

IN VITRO STUDY ON THE CYTOTOXICITY OF THE DIFFERENTLY STERILIZED DEMINERALIZED BONE POWDER (멸균방법에 따른 탈회동종골의 세포독성 여부에 관한 실험적 연구)

  • Woo, Ki-Sun;Yim, Chang-Joon;Kim, Se-Won;Kim, Jong-Yeo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.19 no.3
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    • pp.287-299
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    • 1997
  • Procurement, cutting, cleansing, freezing, freeze-drying, and demineralization of the allogeneic bone must be made under the germ-free stable condition without bacterial and/or viral contamination. Even thought the bone is procured under the germ free condition, we must have confidence on disinfection of all the solutions that come in contact with tissue during the whole procedure. Lots of antibacterial agents have been introduced for chemical sterilization. Recently ethylene oxide gas sterilization or radiation sterilization is frequently selected as a secendary sterilization procedure. The biological and biochemical response of the graft material differs with the type and concentration of the sterilizing agents, and various toxic reactions have been reported due to the graft material itself and the substance released by the chemicals. The authors conducted the Millipore filter test to observe the toxic effect on L929 fibroblasts according to the effect on activity of succinate dehydrogenase, during the secondary sterilization of the demineralized allogeneic bone powder with irradiation or ethylene oxide gas. The result were as follows : 1. Around the copper disk, positive control group, 10mm diameter discoloration was observed. 2. As same as the negative control group, the disk showed no discoloration. 3. The demineralized allogeneic bone which was sterilized with ethylene oxide gas or irradiation showed no cytotoxicity. 4. From this results, it is suggested that treatment with ethylene oxide gas or irradiation should be effective to sterilize the deminineralized allogeneic bone.

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Genotoxicological Safety of Hot Water Extracts of the γ-Irradiated Astragali Radix, Atractylodes Rhizoma, and Cimicifugae Rhizoma in Vitro (감마선 조사 황기, 백출 및 승마 열수 추출물의 in vitro 유전독성학적 안전성 평가)

  • 박혜란;함연호;정우희;정일윤;조성기
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.910-916
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    • 2002
  • As the utilization of medicinal herbs in food and bio-industry increases, safe hygienic technologies for them are demanded. To consider the possibility of application of radiation technology for this purpose, the genotoxi-cological safety of three r -irradiated medicinal herbs were studied. Astragali Radix, Atractylodes Rhizoma and Cimicifugae Rhizoma were irradiated at 10 kGy, and then were extracted with hot water. The genotoxicity of the extracts was examined in two short-term in vitro tests: (1) Salmonella reversion assay (Ames test) in strains of TA98 and TA100; (2) Micronucleus test in cultured Chinese hamster ovary (CHO) cells. The extract was treated at maximum doses of 5 mg/plate in Salmonella reversion assay, and 1 mg/mL in micronucleus test where growth of CHO cells was inhibited by 50%. In Salmonella reversion assay with or without metabolic activation, both ex-tracts of irradiated and non-irradiated herbs showed no significant differences in formation of revertant colonies compared with the negative control. And also in micronucleus test, the incidences of micronucleus in CHO cells cultured with extracts of irradiated herbs were almost same as negative control in less than 3%. These results of two in vitro tests suggest that ${\gamma}$-irradiated herbs do not show mutagenicity and cytogenetic toxicity. Further tests of in vivo genotoxicity and chronic toxicity are needed to ascertain the safety of ${\gamma}$-irradiated herbs.

Selection of Azetidine-2-carboxylic Acid Resistant Cell Lines by in vitro Mutagenesis in Rice (Oryza sativa L.)

  • Hyun, Do-Yoon;Lee, In-Sok;Kim, Dong-Sub;Lee, Sang-Jae;Seo, Yong-Weon;Lee, Young-Il
    • Journal of Plant Biotechnology
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    • v.5 no.1
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    • pp.43-49
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    • 2003
  • Resistant cell lines to azetidine-2-carboxylic acid (AZCA) were selected through rice embryo culture after mutagenic treatment of callus irradiated with 30,50,70,90 and 120 Gy. The optimum AZCA concentration for the selection of resistant cell lines was 3 or 4 mM AZCA considering $LD_{50}$ and the fresh weight of callus. Survival rate of the AZCA resistant callus showed remarkable increase in the callus irradiated with 50 and 70 Gy. Regeneration rate of the AZCA resistant callus was much lower on the whole. Ninety and 120 Gy increased the regeneration rate for calli selected from 3 and 4 mM AZCA, respectively. Based on fresh weight, survival rate and regeneration for selection of the AZCA resistant cell line, 50-90 Gy was considered as the optimum range of gamma irradiation. Irradiated calli selected from AZCA were more tolerant to NaCl than those from non-irradiated calli. It suggests that elevated resistance to osmotic stress resulted from mutagenic treatment. The level of free proline content in the AZCA resistant cell line was increased up to 3.5 times compared with that in the control. Proline content in the regenerant derived from the AZCA resistant cell line also increased to 1.7 times that from the control plants regenerated from callus grown in AZCA free medium. Selection of proline overproducing cell lines by in vitro mutagenesis was successful and seems to be useful for improvement of stress tolerance in this crop.

Phototoxicity: Its Mechanism and Animal Alternative Test Methods

  • Kim, Kyuri;Park, Hyeonji;Lim, Kyung-Min
    • Toxicological Research
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    • v.31 no.2
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    • pp.97-104
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    • 2015
  • The skin exposure to solar irradiation and photoreactive xenobiotics may produce abnormal skin reaction, phototoxicity. Phototoxicity is an acute light-induced response, which occurs when photoreacive chemicals are activated by solar lights and transformed into products cytotoxic against the skin cells. Multifarious symptoms of phototoxicity are identified, skin irritation, erythema, pruritis, and edema that are similar to those of the exaggerated sunburn. Diverse organic chemicals, especially drugs, are known to induce phototoxicity, which is probably from the common possession of UV-absorbing benzene or heterocyclic rings in their molecular structures. Both UVB (290~320 nm) and UVA (320~400 nm) are responsible for the manifestation of phototoxicity. Absorption of photons and absorbed energy (hv) by photoactive chemicals results in molecular changes or generates reactive oxygen species and depending on the way how endogenous molecules are affected by phototoxicants, mechanisms of phototoxcity is categorized into two modes of action: Direct when unstable species from excited state directly react with the endogenous molecules, and indirect when endogeneous molecules react with secondary photoproducts. In order to identify phototoxic potential of a chemical, various test methods have been introduced. Focus is given to animal alternative test methods, i.e., in vitro, and in chemico assays as well as in vivo. 3T3 neutral red uptake assay, erythrocyte photohemolysis test, and phototoxicity test using human 3-dimensional (3D) epidermis model are examples of in vitro assays. In chemico methods evaluate the generation of reactive oxygen species or DNA strand break activity employing plasmid for chemicals, or drugs with phototoxic potential.

Induction of Mutants by Irradiation of $\gamma$-Ray on In vitro Shoots of Persimmon (기내$\gamma$-선 처리에 의한 감(Diospyros kaki Thunb.) 돌연변이 유기)

  • 고갑천
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.2
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    • pp.143-148
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    • 2000
  • This study was carried out to know the optimum dose of gamma-ray for the induction of mutation in vitro and the characteristics of the mutants induced by gamma-ray in persimmon (Diospyros kaki Thunb.). The LD50 (50% lethal dose) for in vitro shoots of the cultivar, Nishimurawase was between 1 krad and 2 krad and about 1 krad for the cultivar, Ichikikeijiro. As the dose of gamma-ray increased, the length of shoots decreased and necrosis of buds increased. For the cultivar, Nishmurawase, 37.5∼58.3% shoots rooted and the rooting rate and the number of roots per shoot was low in high gamma-ray. The irradiated young plants which were grown in the growth cabinet for 6 weeks were shorter in shoot length and had more branches than non-irradiated plants. The survival rate of irradiated plants grown in the green house for 3 months was 33%, while 77% for control plants.

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INFLUENCE OF LIGHT IRRADIATION OVER SELF-PRIMING ADHESIVE ON DENTIN BONDING (상아질접착제에 대한 광조사가 접착에 미치는 영향)

  • 류현욱;김기옥;김성교
    • Restorative Dentistry and Endodontics
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    • v.26 no.5
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    • pp.409-417
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    • 2001
  • The purpose of this study was to investigate the influence of light irradiation over self-priming adhesive on dentin bonding. After acid etching the exposed dentin, a self-priming adhesive (Prime&Bond$^{\circledR}$NT dental adhesive system Dentsply DeTrey, GmbH, Konstanz, Germany) was applied and light irradiation was done for 20 sec with regular intensity (600 mW/$\textrm{cm}^2$) in group I and for 3 sec with ultra-high intensity (1930 mW/$\textrm{cm}^2$) in group III. No light irradiation was done over self-priming adhesive in groups II and IV. Composite resin was added on the self-priming adhesive and irradiated for 40 sec with regular intensity (600 mW/$\textrm{cm}^2$) in groups I and II and for 3 sec with ultra-high intensity (1930 mW/$\textrm{cm}^2$) in groups III and IV. To see the effect of light curing time on dentin bonding, another 3 group specimens were prepared. Without light-irradiation over self-priming adhesive, added composite resin was irradiated for 3, 6, or 12 sec with ultra-high intensity light. After bonded specimens were stored in 37$^{\circ}C$ distilled water for 24 hours, shear bond strength were measured using a universal testing machine (4202, Instron, Instron Co., U.S.A.) and fractured surfaces were examined under a stereomicroscope (SZ-PT Olympus, Japan). Statistical analysis were done with one-way, two-way ANOVA and chi-square test. The results were as follows : 1. The shear bond strengths from the groups irradiated over self-priming adhesive were significantly higher than those from the groups without irradiation (p<0.05). 2. There was no significant shear bond strength difference between regular intensity light irradiation groups and ultra-high intensity ones (p>0.05). 3. There was no significant shear bond strength difference among various irradiation time groups with ultra-high intensity ones (p>0.05). 4. In stereomicroscopic examination of fractured surfaces, adhesive-cohesive mixed failure mode was mostly seen in all groups, and there was no significant difference in failure mode among groups (p>0.05).

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Effects of Gamma Irradiation on Color Changes and Antioxidative Activities of Caesalpinia sappan L. (감마선 조사에 의한 소목추출물의 색상변화 및 항산화 활성 평가)

  • Kwon, Hyung-Jun;Jung, U-Hee;Park, Hae-Ran;Shin, Dong-Hwa;Jo, Sung-Kee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.8
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    • pp.1055-1061
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    • 2007
  • Caesalpinia sappan L. is an indeciduous tree distributed in China, India, Miyanmar and Vietnam. Its heartwood has long been used in oriental folk medicines to treat diseases. In this study, antioxidative activities of Caesalpinia sappan L. and the effect of gamma irradiation on its chemical and biological properties were investigated. The ethyl acetate fraction (EtOAc fr.) of Caesalpinia sappan L. was irradiated with 100 kGy of gamma ray. The dark red color of EtOAc fr. was significantly (p<0.05) removed by irradiation (Hunter L and b values increased and a value decreased). The total phenolic content of EtOAc fr. was 865 mg/g and it was increased to 1195 mg/g by gamma irradiation. DPPH radical and superoxide anion radical scavenging activities and lipid peroxidation inhibitory activity of EtOAc fr. were very high and its activities were also increased by gamma irradiation. EtOAc fr. also inhibited the irradiation-induced DNA damage of lymphocyte as determined by comet assay. In conclusion, EtOAc fr. of Caesalpinia sappan L. extract showed high antioxidative activities in vitro. Furthermore, gamma irradiation on EtOAc fr. ameliorated the color and antioxidative properties. Therefore, it can be suggested that Caesalpinia sappan L. may be a good material for antioxidant function and gamma irradiation may be applied for the improvement of chemical and biological properties of Caesalpinia sappan L.