• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.29 no.2
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• pp.435-449
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• 1999

Purpose : The study was aimed to detect the differences in the cell viability and the apoptosis induction after irradiation on normal and tumorigenic cells. Materials and Methods : The study. that was generated for two human normal cells(RHEK, HGF-l) and two human tumor cells(KB. HT-1080). was tested using MTT assay at 1 day and 3 day after irradiation and TUNEL assay under confocal laser scanning microscope at 1 day after irradiation. Single irradiation of 0.5. 1, 2. 4. and 8Gy were applied to the cells. The two fractions of 1. 2. 4. and 8Gy were separated with a 4-hour time interval. The irradiation was done with 5.38Gy/min dose rate using Cs-137 irradiator at room temperature. Results and Conclusions : 1. In 3-day group. the cell viability of HGF-1 cell was significantly decreased at 2. 4 and 8Gy irradiation, the cell viability of KB cell was significantly decreased at 8Gy irradiation and the cell viability of HT-I080 cell was significantly decreased at 4 and 8Gy irradiation. 2. There was significant difference between RHEK and KB cell line in the cell viability of 3-day group at 8Gy irradiation. There was significant difference between RHEK and HGF-1 cell line in the cell viability of 3-day group at 4 and 8Gy irradiation. 3. There was a significantly decreased cell viability in 3-day group than those in 1-day group at 2. 4 and 8Gy on HGF-1 cell. at 4 and 8Gy on HT-I080 cell. at 8Gy on KB cell. 4. We could detect DNA fragmented cells only on KB cell. Number of apoptotic cells of KB cell was significantly increased at 4 and 8Gy irradiation. However, there was no correlation between cell viability and apoptosis. 5. On all 4 cell lines, there were no differences between single and split irradiation method in cell viability and apoptosis.

• Imaging Science in Dentistry
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• v.35 no.1
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• pp.1-8
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• 2005

Purpose : To investigate the effects of irradiation on the calcium content and calcific nodule formation in the MC3T3-E1 osteoblastic cell line. Materials and Methods : Cells were irradiated with a single dose of 2,4 and 8 Gy at a dose rate of 5.38 Gy/min using a Cs-137 irradiator. After irradiation, the calcium content and calcific nodule formation were examined on the 1 st, 2nd, 3rd and 4th week. Results : A decreasing dose-dependent tendency of the cell proliferation rate was found in all irradiated groups of this experiment when compared with the unirradiated control group. In accordance with the duration of culture, there was no significant difference in the cell proliferation rate after irradiation of 2 Gy when compared with the unirradiated group, however a decreasing tendency was found in 4 Gy- and 8 Gy-irradiated groups. While an increase in total calcium content after irradiation of 2 Gy was found at week 1, week 2, and week 4, there was a decrease in calcium content at week 1 through 4 in the 8 Gy- irradiated group. Calcific nodule formation was increased in irradiated experimental groups when compared with the unirradiated control group in the 2 Gy-irradiated group, but decreased in the 4 Gy- and 8 Gy-irradiated groups at the same stage. Conclusion : The results showed a mild increasing tendency of the calcific nodule formation after irradiation of 2 Gy. However, a decreased calcific nodule formation in 4 Gy- and 8 Gy-irradiated groups was found. Taken together, the irradiation of 2 Gy mildly activated bone formation, however 4 Gy or 8 Gy suppressed bone formation by decreasing cell numbers in the MC3T3-El osteoblastic cell line.

• Imaging Science in Dentistry
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• v.30 no.1
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• pp.71-79
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• 2000

Purpose : This study was aimed to evaluate the cell cycle arrest and apoptosis induction after irradiation and epidermal growth factor (EGF) treatment in three human epithelial tumor cell lines (A431, Siha, KB). Materials and Methods: Single irradiation of 2, 5 and 10 Gy was done on three cell lines with 5.38 Gy/min dose rate using Cs-137 irradiator at room temperature. Also, EGF of 10 ng/ml was added immediately after 10 Gy irradiation. Cell growth was evaluated by counting the living cell number using a hemocytometer at 1 day, 2 days, 3 days, 4 days and 5 days after irradiation. Cell cycle arrest and apoptosis induction were assayed with the flow cytometry at 8 hours, 12 hours, 1 day, 2 days, 3 days, 4 days and 5 days after irradiation. Results : Growth of irradiated three cell lines were inhibited in proportion to radiation dose. EGF treatment after irradiation showed various results according to cell lines. On all cell lines, G2 arrest was detected after 8 hours and maximized after 12 hours or 1 day. Amount of G2 arrest was positively dose dependent. However, EGF showed no significant change on G2 arrest. G2 arrest was recovered with time at 2 Gy and 5 Gy irradiation. However, at 10 Gy irradiation, G2 arrest was continued. Apoptosis was detected at 10 Gy irradiation. On EGF treated group after irradiation, A431 and Siha cell lines showed slightly increased apoptosis but there was no statistically significant difference. KB cell line showed no marked change of apoptosis induction. Conclusion : Irradiation effects on cell cycle arrest and apoptosis induction in three human epithelial tumor cell lines, however epidermal growth factor doesn't effect on.

• Korean Journal of Head & Neck Oncology
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• v.22 no.2
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• pp.130-136
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• 2006

Introduction : The sensitivity of tumor cells to radiotherapy is a critical determinant of local control and potential cure in advanced head and neck squamous cell carcinoma(HNSCC). The emergence of radioresistant tumor cells is an obstacle to cancer therapy. Most radioresistant cells have a higher proportion of cells in the Sphase of the cell cycle and a lower apoptotic fraction than radiosensitive cells. HSV replication is increased in cells that have higher S-phase fractions. NV1066 is an oncolytic herpes simplex virus type-1 mutant. We hypothesized that NV1066 replication and cytotoxicity are increased in radioresistant cells. The purpose of this study is to evaluate the antitumor efficacy of NV1066 to treat radioresistant HNSCC. Methods : Radioresistant cells were selected by treating five HNSCC cell lines with repeated conventional fractionated doses of radiation(2Gy/day), using a Cs-137 irradiator, up to a cumulative dose of 70Gy. Clonogenic cell survival and S-phase fractions were compared between radioresistant and parental radiosensitive cells. The two cell populations were then treated with NV1066 to examine viral replication, by the viral plaque assay and viral cytotoxicity. Results : Fractionated irradiation resulted in the selection of radioresistant cells. Radioresistant cells had a higher S-phase fraction(42.9%) compared to parental cells(26.2%). NV1066 replication in radioresistant cells was 7.4 times higher than in parental cells(p<0.01). Treatment with NV1066 resulted in increased cytotoxicity of 24.5% in radioresistant cells compared to parental cells(p<0.05). Conclusion : NV1066 showed increased viral replication and cytotoxicity in radioresistant HNSCC cell lines. These findings suggest a potential clinical application for this oncolytic viral therapy as treatment for radioresistant head and neck cancers.

• Imaging Science in Dentistry
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• v.30 no.4
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• pp.275-279
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• 2000

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 /sup 137/Cs 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 flowcytometry (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.

• The Korean Journal of Nuclear Medicine
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• v.33 no.2
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• pp.172-177
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• 1999

Purpose: The purpose of this study was to ascertain whether radiation adaptive response could be induced by Tc-99m-methylene diphosphonate (Tc-99m-MDP) in peripheral lymphocytes of patients undergoing bone scintigraphy. Materials and Methods: Lymphocytes from 22 patients (6 males, 16 females, mean age $50{\pm}14$ years) were collected before and after bone scintigraphy using 740 MBq Tc-99m-MDP Lymphocytes from 10 controls (6 males, 4 females, mean age $43{\pm}7$ years) were also collected. They were exposed to challenge dose of 2 Gy gamma rays using a cell irradiator. Number of ring-form and dicentric chromosomes per 600 cells (chromosomal aberrations) was counted under the light microscope. Results: Chromosomal aberrations in patients before bone scintigraphy ($385.1{\pm}30.5$) was not different from that of controls ($367.8{\pm}36.6$). However, chromosomal aberrations in patients after bone scintigraphy was significantly decreased to $192.6{\pm}22.1$ (p=0.0001). Conclusion: Low dose gamma-irradiation by Tc-99m-MDP used for bone scintigraphy induces a cytogenetic adaptive response in peripheral lymphocytes.

• Journal of Radiation Protection and Research
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• v.28 no.2
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• pp.117-125
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• 2003

It was reported that radiopharamaceuticals induced radiation adaptive response (RAR) in patients undergoing nuclear medicine imaging studies. Individual variations of RAR were not studied well. The purpose of this study was to evaluate individual variation of RAR in patients undergoing nuclear medicine imaging studies. Peripheral lymphocytes were collected from 23 patients undergoing $^{99m}Tc-diethylenetriamine$ pentaacetic acid $(^{99m}Tc-DTPA)$ renal scintigraphy, 18 patients undergoing $^{99m}Tc-methylene$ diphosphonate $(^{99m}Tc-MDP)$ bone scintigraphy and 21 patients undergoing $^{99m}Tc-tetrofosmin\;(^{99m}Tc-TF)$ scintigraphy were collected before and 4 hours after injection of radiopharmaceuticals. The lymphocytes were exposed to challenge dose of 2 Gy gamma rays using a cell irradiator. Numbers of ring-form (R) and dicentric (D) chromosomes were counted under the light microscope. and used to calculate the frequency of chromosomal aberration [Ydr=(D+R)/total number of counted lymphocytes]. Adaptation index (k) was defined 3s ratio of Ydr in conditioned lymphocytes over Ydr in unconditioned lymphocytes. Coefficients of variance of k in $^{99m}Tc-DTPA,\;^{99m}Tc-MDP\;and\;^{99m}Tc-TF$ were 35%, 34% and 21%, respectively k was not dependent upon age, sex, and underlying diseases. There was a wide variation of RAR induced by radiopharmaceuticals among patients undergoing nuclear medicine procedures. It remains to be determined for causes of such variation.

• The Korean Journal of Nuclear Medicine
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• v.33 no.3
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• pp.306-315
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• 1999

Purpose: We performed this study to evaluate the process of radiation induced apoptosis in A431 skin epithelial cancer cell line. Materials and Methods: Low to high dose radiation (0, 2, 5, 10, 25 Gy) was given to A431 cells by Cs-137 cell irradiator. Apoptosis was evaluated by cell morphology, dye exclusion test, and DNA laddering. Results: Cell viability decreased as the radiation dose increased. Number of apoptotic bodies increased as radiation dose increased. It increased most significantly at 12 hours after irradiation. Lactate dehydrogenase activity in culture medium increased according to radiation dose and time after irradiation. DNA ladders could be identified in irradiated cells, but, it had no correlation with radiation dose or time after irradiation. Conclusion: Radiation-induced apoptosis which was the main course of cell death in A431 cells could be analyzed quantitatively by counting apoptotic bodies under microscope. Apoptosis increased as radiation dose increased.

• The Korean Journal of Nuclear Medicine
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• v.34 no.3
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• pp.252-259
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• 2000

Purpose: The purpose of this study was to compare the radiation adaptive response (RAR) in peripheral lymphocytes (PL) of patients induced by Tc-99m MDP and Tc-99m DTPA scintigraphies. Materials and Methods: Lymphocytes from 45 patients (25 males, 20 females, mean age $44{\pm}18$ years) were collected before and after scintigraphies using 740 MBq Tc-99m MDP (n=22) or Tc-99m DTPA (n=23). Lymphocytes from 20 controls (12 males, 8 females, mean age $43{\pm}7$ years) were also collected. They were exposed to challenge dose of 2 Gy ${\gamma}-rays$ using a Cs-137 cell irradiator Number of ring-form (R) and dicentric (D) chromosomes was counted under the light microscope. From them a representative score, Ydr, was calculated as Ydr=(D+R)/cells. Adaptation index (AI) was defined as difference of Ydr between unconditioned and conditioned lymphocytes. Ydr was also measured after an administration of cycloheximide (CHM), a protein synthesis inhibitor, before challenge dose. Results: RAR was induced in both groups of patients. CHM abolished the adaptive response in both groups. AI of Tc-99m MDP group was significantly higher than that of Tc-99m BTPA group. Conclusion: Tc-99m MDP induced RAR was more prominent than those induced by Tc-99m DTPA.

• The Korean Journal of Nuclear Medicine
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• v.35 no.2
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• pp.83-88
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• 2001

Purpose: The purpose of this study was to ascertain whether radiation adaptive response could be induced by high dose I-131 therapy in patients with differentiated thyroid cancer. Materials and Methods: Lymphocytes from 21 patients (7 males, 14 females, mean age $55{\pm}12$ years) were collected before and after administration of 5,550 MBq (150 mCi) I-131. They were exposed to a challenge dose of 1 Gy gamma rays using a Cs-137 cell irradiator. The number of ring-form (R) and dicentric (D) chromosomes was counted under the light microscope, and used to calculate the frequency of chromosomal aberration. Ydr, which was defined as the sum of R and D divided by the total number of counted lymphocytes. Results: Ydr in patients before I-131 therapy ($0.09{\pm}0.01$) was not different from that of controls ($0.08{\pm}0.01$). Ydr was significantly increased to $0.13{\pm}0.02$ (p<0.0001) after I-131 therapy. Increase of Ydr after the challenge irradiation of 1 Gy was significantly lower in patients after I-131 therapy than before I-131 therapy ($0.17{\pm}0.03\;vs\;0.21{\pm}0.02$, p<0.0001). Cycloheximide (CHM), an inhibitor of protein synthesis, abolished this effect. Ydr after CHM ($0.20{\pm}0.01$) was significantly higher than Ydr after I-131 therapy ($0.17{\pm}0.03$, p<0.0001), but was not different from Ydr before I-131 therapy ($0.21{\pm}0.02$).Conclusion: High dose I-131 therapy induces an adaptive response in peripheral lymphocytes of patients with well-differentiated thyroid cancer, which is associated with protein synthesis.