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Radiation-induced Apoptosis is Differentially Modulated by PTK Inhibitors in K562 Cells (K562 백혈병 세포주에서 방사선에 의해 유도되는 Apoptosis에 미치는 PTK Inhibitors의 영향)

  • Lee Hyung Sik;Moon Chang Woo;Hur Won Joo;Jeong Su Jin;Jeong Min Ho;Lee Jeong Hyeon;Lim Young kin;Park Heon Joo
    • Radiation Oncology Journal
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    • v.18 no.1
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    • pp.51-58
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    • 2000
  • Purpose :The effect of PTK inhibitors (herbimycin A and genistein) on the induction of radiation-induced apoptosis in Ph-positive KS62 leukemia cell line was investigated. Materials and Methods :K562 cells in exponential growth phase were irradiated with a linear accelerator at room temperature. For 6 MV X-ray irradiation and drug treatment, cultures were initiated at 2×106 cells/mL. The cells were irradiated with 10 Gy. Stock solutions of herbimycin A and genistein were prepared in dimethyl sulphoxide (DMSO). After incubation at 37$^{\circ}C$ for 0$\~$48 h, the extent of apoptosis was determined using agarose gel electrophoresis and TUNEL assay. The progression of cells through the cell cycle after irradiation and drug treatment was also determined with flow cytometry. Western blot analysis was used to monitor bel-2, bel-X$_{L}$ and bax protein levels. Results :Treatment with 10 Gy X-irradiation did not result in the induction of apoptosis. The HMA alone (500 nM) also failed to induce apoptosis. By contrast, incubation of K562 cells with HMA after irradiation resulted in a substantial induction of nuclear condensation and fragmentation by agarose gel electro-phoresis and TUNEL assay. Genistein failed to enhance the ability of X-irradiation to induce DNA fragmentation. Enhancement of apoptosis by HMA was not attributable to downregulation of the bel-2 or bel-X$_{L}$ anti-apoptotic proteins. When the cells were irradiated and maintained with HMA, the percentage of cells in G2/M phase decreased to 30$\~$40$\%$ at 48 h. On the other hand, cells exposed to 10 Gy X-irradiation alone or maintained with genistein did not show marked cell cycle redistribution. Conclusion : We have shown that nanomolar concentrations of the PTK inhibitor HMA synergize with X-irradiation in inducing the apoptosis in Ph (+) K562 leukemia cell line. While, genistein, a PTK inhibitor which is not selective for p210$^{bcr/abl}$ failed to enhance the radiation induced apoptosis in KS62 cells. It is unlikely that the ability of HMA to enhance apoptosis in K562 cells is attributable to bel-2 family. It is plausible that the relationship between cell cycle delays and cell death is essential for drug development based on molecular targeting designed to modify radiation-induced apoptosis.

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A Novel Chenodeoxycholic Derivative HS-1200 Induces Apoptosis in Human HT-29 Colon Cancer Cells (인체 대장암 세포주(HT-29)에서 담즙산 합성유도체(HS-1200)의 세포 사망 기전)

  • Oh Sin Geun;Yang Kwang Mo;Hur Won Joo;Yoo Young Hyun;Suh Hong Suk;Lee Hyung Sik
    • Radiation Oncology Journal
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    • v.20 no.4
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    • pp.367-374
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    • 2002
  • Purpose : To investigate the growth inhibitory effects, and the underlying mechanism of human colon cancer cell (HT-29) death, induced by a new synthetic bile acid derivative (HS-1200). Materials and Methods : Human colon cancer cells (HT-29), in exponential growth phase, were treated with various concentrations of a new synthetic bile acid derivative (HS-1200). The growth inhibitory effects on HT-29 cells were examined using a frypan blue exclusion assay. The extent of apoptosis was determined using agarose gel electrophoresis, TUNEL assays and Hoechst staining. The apoptotic cell death was also confirmed by Western blotting of PARP, caspase-3 and DNA fragmentation factor (DFF) analysis. To investigate the involvement of mitochondria, we employed immunofluorescent staining of cytochrome c and mitochondrial membrane potential analyses. Results : The dose required for the half maximal inhibition $(IC_{50})$ of the HT-29 cell growth was $100\~150\;{\mu}M$ of HS-1200. Several changes, associated with the apoptosis of the HT-29 cells, were reveal by the agarose gel eletrophoresis, TUNEL assays and Hoechst staining, following their treatment with $100\;{\mu}M$ of HS-1200. HS-1200 treatment also induced caspase-3, PARP and DFF degradations, and the western blotting showed the processed caspase-3 p20, PARP p85 and DFF p30 and p11 cleaved products. Mitochondrial events were also demonstrated. The cytochrome c staining indicated that cytochrome c had been released from the mitochondria in the HS-1200 treated cells. The mitochondrial membrane potential $(\Delta\Psi_m)$ was also prominently decreased in the HS-1200 treated cells. Conclusion : These findings suggest that the HS-1200 - induced apoptosis of human colon cancer cells (HT-29) is mediated via caspase and mitochondrial pathways.

A Novel Chenodeoxycholic Derivative HS-1200 Enhances Radiation-induced Apoptosis in Human MCF-7 Breast Cancer Cells (담즙산 합성유도체(HS-1200)가 인체 유방암 세포주(MCF-7)에서 유도하는 방사선 감작 효과)

  • Lee Hyung Sik;Choi Young Min;Kwon Hyuk Chan;Song Yeon Suk
    • Radiation Oncology Journal
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    • v.22 no.2
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    • pp.145-154
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    • 2004
  • Purpose : To examine whether a synthetic bile acid derivatives (HS-1200) sensitizes the radiation-induced apoptosis in human breast cancer cells (MCF-7) and to investigate the underlying mechanism. Materials and Methods : Human breast cancer cells (MCF-7) in exponential growth phase were treated with HS-1200 for 24 hours at 37$^{\circ}C$ with 5$\%$ CO$_{2}$ in air atmosphere. After removal of HS-1200, cells were irradiated with 2$\~$8 Gy X-ray, and then cultured Ii drug-free media for 24-96 hours. The effect of radiation on the clonogenicity of MCF-7 cells was determined with clonogenic cell survival assay with 16$\mu$M of HS-1200. The induction of apoptosis was determined using agarose gel electrophoresis and Hoechst staining. The expression level of apoptosis-related molecules, such as PARP, Bax, Bcl-2, Bak and AIF, were assayed by Western blotting analysis with 40$\mu$M of HS-1200 combined with 8 Gy irradiation. To examine the cellular location of cytochrome c, bax and AIF immunofluorescent stainings were undertaken. Results : Treatment of MCF-7 cells with 40$\mu$M of HS-1200 combined with 8 Gy irradiation showed several changes associated with enhanced apoptosis by agarose gel electrophoresis and Hoechst staining. HS-1200 combined with 8 Gy irradiation treatment also enhanced production of PARP cleavage products and increased Bax/Bcl-2 ratio by Western blotting. Loss of mitochondrial membrane potential ($\Delta$$\psi$$_{m}$) and increased cytochrome c staining indicated that cytochrome c had been released from the mitochondria in HS-1200 treated cells. Conclusion : We demonstrated that combination treatment with a synthetic chenodeoxycholic acid derivative HS-1200 and irradiation enhanced radiation-induced apoptosis of human breast cancer cells (MCF-7). We suggest that the increased Bax/Bcl-2 ratio In HS-1200 co-treatment group underlies the increased radio sensitivity of MCF-7 cells. Further futures studies are remained elusive.

The effects of different intensity endurance and resistance exercise on diabetic-related blood profiles in impaired glucose tolerance mice (다른 운동 강도의 지구성 및 저항성 운동 중재가 내당능 장애 쥐들의 당뇨관련 혈액인자에 미치는 영향)

  • Park, Chan-Ho;Woo, Jin-Hee;Roh, Hee-Tae;Shin, Ki-Ok;Kim, Do-Yeon;Yoon, Byung-Kon
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.3
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    • pp.571-581
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    • 2020
  • The purpose of this study was to investigate the effect of endurance and resistance exercise by using different intensity on diabetic-related blood profiles in impaired glucose tolerance mice. 54 C57BL/6 mice were divided into (1) Control group (CO, n=9), (2) impaired glucose tolerance group (IGT, n=9), (3) IGT + 50% of VO2max endurance exercise group (IGT50A, n=9), (4) IGT + 75% of VO2max endurance exercise group (IGT75A, n=9), (5) IGT +50% of 1RM resistance exercise group (IGT50R, n=9), 그리고 (6) IGT + 75% of 1RM resistance exercise group (IGT75R, n=9). Endurance exercise program was performed 40 min/day, 5 days a week for 8 weeks on treadmill. Resistance exercise program was consisted in ladder-climbing 8 set/day, 5 days a week for 8 weeks. After completed the exercise program, there was no different insulin level in the groups. The fasting glucose was significantly lower in CO than in other groups, and it was not different among the exercised groups. However, IGT75R was statistically lower than IGT. HOMA-IR was only different between CO and IGT. HbA1c was higher in IGT than in other groups. As compared with exercised groups, IGT 75A was lower than IGT50R. TC was lower in CO than in other groups, but there was no different in TG. Endurance exercise groups showed higher than CO and IGT in HDL-C level. LDL-C was lower in CO than other groups. In addition, IGT75A was lower than IGT and IGT50R in LDL-C level. In conclusion, 75% of 1RM resistance exercise had more positive effect on fasting glucose, and 75% of VO2max endurance exercise improved HbA1c and LDL-C level. In addition, endurance exercise had more effective in HDL-C improvement as compared with resistance exercise.

Differential Levels of Governance and Its Impact on Urban Park Management and Users' Satisfaction - The Case of Sheffield District Parks, UK - (도시공원관리 거버넌스 구축정도에 따른 이용자 만족도 차이 - 영국 셰필드 지구공원을 대상으로 -)

  • Nam, Jinvo;Kim, Hyun
    • Journal of the Korean Institute of Landscape Architecture
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    • v.47 no.4
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    • pp.50-60
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    • 2019
  • In the late 1980s, a financial crisis and Compulsory Competitive Tendering (CCT) in green space services brought with it a profound impact on the quality of parks in the UK. Such government projects, e.g. Urban Task Force (1999) and Public Parks Assessment (2001), aimed to raise the awareness of the severity of the declining standards of urban parks. Since the late 1990s, the UK governments (The New Labour (1997-2010) and The Conservative Government (2010-2019)), have often adopted community-led governance schemes to enhance the quality of parks and address problems derived from the financial crisis. Accordingly, community groups, notably 'Friends of', enlarged their involvement in the decision-making process of park management. However, there is little empirical evidence concerning the impact of community-led governance on park management, in particular, the effect on the users' perceptions of park use. This study explored the context of community-led park management to reclassify the level of build-up of governance underlined by 'A Ladder of Citizen Participation'. In addition, questionnaire surveys were conducted around two Sheffield district parks, which are located in deprived areas. As a result, community involvement in the status quo of UK urban park management has changed its form of governance based on the extent of involvement in the decision-making process. The forms of governance could be categorised in three levels: general, active, and predominant governance, where the extents of decision-making and sharing responsibility vary. The results obtained through the questionnaires show that one park (active governance), which has a stronger tendency of sharing responsibility to get involved in park management, had better contribution to park management and positive impacts on users' satisfaction than the other park (general governance). The findings highlight that stronger governance in partnerships with the non-public sectors can shed light on current and future park management through a shift in sharing responsibility for park management.

Swimming Performance Evaluation of Four Freshwater Fish Species from the South Korea (국내에 서식하는 담수어류 4종에 대한 유영능력 평가)

  • Misheel, Bold;Kim, Kyu-Jin;Min, Kun-Woo;Jang, Min-Ho
    • Korean Journal of Ecology and Environment
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    • v.52 no.2
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    • pp.118-125
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    • 2019
  • Swimming performance of fish is an important factor in the survival of fish. Also, swimming performance of fish is used in the form of habitat, or as a condition to consider when creating a fish ladder. However in Korea, researches in swimming performance of Korean freshwater fish were scarce and inadequate in some part, thus fish ladders were installed without considering their swimming performance. Therefore, in this study, we measured swimming performance of 4 Korean freshwater fish species to consider importance of swimming performance test. The fish used in this study were Carassius auratus, Zacco koreanus, Gnathopogon strigatus, Acheilognathus lanceolata intermedia species which was collected during October to November, 2018 at Geum River, and measurement for swimming speed of each fish was done by using $Loligo^{(R)}$ System, swim tunnel respirometer in January to February of 2019. The average value of the burst critical swimming speed ($U_{crit}$) for each species was $0.8{\pm}0.04m\;s^{-1}$ for C. auratus, $0.77{\pm}0.04m\;s^{-1}$ for Z. koreanus, $0.95{\pm}0.04m\;s^{-1}$ for G. strigatus, $0.73{\pm}0.03m\;s^{-1}$ for A. lanceolata intermedia and the average value of prolonged critical swimming speed was $0.54m\;s^{-1}$ for C. auratus, $0.67m\;s^{-1}$ for Z. koreanus, $0.7m\;s^{-1}$ for G. strigatus, $0.54m\;s^{-1}$ for A. lanceolata intermedia. Since the fish used in this experiment were collected from a small part of the water system in Korea and there were only 4 species, they were not enough to represent the species that inhabit the entire Korean water system. It will be necessary to continue evaluating the swimming performance of other freshwater species in Korea.