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      • SCOPUSKCI등재

        Radiation-induced Apoptosis, Necrosis and G2 Arrest in Fadu and Hep2 Cells

        Lee, Sam-Sun,Kang, Beom-Hyun,Choi, Hang-Moon,Jeon, In-Seong,Heo, Min-Suk,Choi, Soon-Chul 大韓口腔顎顔面 放射線學會 2000 Imaging Science in Dentistry Vol.30 No.4

        Purpose : Radiation damage is produced and viable cell number is reduced. We need to know the type of cell death by the ionizing radiation and the amount and duration of cell cycle arrest. In this study, we want to identified the main cause of the cellular damage in the oral cancer cells and normal keratinocytes with clinically useful radiation dosage. Materials and Methods : Human gingival tissue specimens obtained from healthy volunteers were used for primary culture of the normal human oral keratinocytes(NHOK). Primary NHOK were prepared from separated epithelial tissue and maintained in keratinocyte growth medium containing 0.15 mM calcium and a supplementary growth factor bullet kit. Fadu and Hep-2 cell lines were obtained from KCLB, Cells were irradiated in a 137Cs γ-irradiator at the dose of 10 Gy. The dose rate was 5.38 Gy/min. The necrotic cell death was examined with Lactate Dehydrogenase(LDH) activity in the culture medium. Every 4 day after irradiation, LDH activities were read and compared control group. Cell cycle phase distribution and preG1-incidence after radiation were analyzed by flow cytometry using Propidium Iodine staining. Cell cycle analysis were carried out with a FAC Star plus flowcytome-try (FACS , Becton Dickinson, USA) and DNA histograms were processed with CELLFIT software (Becton Dickinson, USA). Results : LDH activity increased in all of the experimental cells by the times. This pattern could be seen in the non-irradiated cells, and there was no difference between the non-irradiated cells and irradiated cells. We detected an induction of apoptosis after irradiation with a single dose of 10 Gy. The maximal rate of apoptosis ranged from 4.0% to 8.0% 4 days after irradiation. In all experimental cells, we detected G2/M arrest after irradiation with a single dose of 10 Gy. Yet there were differences in the number of G2/M arrested cells, The maximal rate of the G2/M ranges from 60.0% to 80.0% 24h after irradiation. There is no significant changes on the rate of the G0/G1 phase. Conclusion : Radiation sensitivity was not related with necrosis but cell cycle arrest and apoptosis. These data suggested that more arrested cell is correlated with more apoptosis. (Korean J Oral Maxillofac Radiol 2000;30:275-279)

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        두경부 편평세포암종세포주에서 retinoic acid가 linear-quadratic 모델을 적용한 방사선감수성과 apoptosis 유발에 미치는 영향

        이은숙,강범현,허민석,이삼선,최현배,최순철,박태원 대한구강악안면방사선학회 2001 Imaging Science in Dentistry Vol.31 No.3

        Purpose : To evaluate the effect of all-trans-retinoic acid on radiosensitivity and radiation-induced apoptosis in NHOK, HEp-2 and FaDu cell lines. Material and Methods : We measured the changes in survival fraction at 2 Gy (SF2), α and β after treatment of retinoic acid (1μM) prior to irradiation with doses of 2,4,6 and 10 Gy and correlated the radiosensitizing effect of retinoic acid with them. Also, apoptosis induction was assayed with the flow cytometry on days 1, 2, 3, 4 and 5 after irradiation (2,10 and 20 Gy) combined with retinoic acid. Results and Conclusion : SF2 values for NHOK, HEp-2 and FaDu cell lines were 0.54, 0.64 and 0.41, respectively and the cell line of FaDu was the most radiosensitive, For cell lines of NHOK and HEp-2, pretreatment of cells with retinoic acid resulted in a significant decrease of the SF2 values. The α/β ratios of x-ray survival curve were 8.714(NHOK), 4.098(HEp-2) and 11.79(FaDu). The α/β ratio for NHOK decreased on pretreatment with retinoic acid, whereas those for HEp-2 and FaDu increased. Radiation induced apoptosis in all cell lines but, retinoic acid did not affect the apoptosis. (Korean J Oral Maxillofac Radiol 2001; 31 : 135-43)

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