To study effects of cellular FLICE (FADD-like IL-$1{\beta}$-converting enzyme)-inhibitory protein (c-FLIP) inhibition by RNA interference (RNAi) on sensitivity of U2OS cells to tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL)-induced apoptosis, plasmid pSUPER-c-FLIP-siRNA was constructed and then transfected into U2OS cells. A stable transfection cell clone U2OS/pSUPER-c-FLIP-siRNA was screened from the c-FLIP-siRNA transfected cells. RT-PCR and Western blotting were applied to measure the expression of c-FLIP at the levels of mRNA and protein. The results indicated that the expression of c-FLIP was significantly suppressed by the c-FLIP-siRNA in the cloned U2OS/pSUPER-c-FLIP-siRNA as compared with the control cells of U2OS/pSUPER. The cloned cell line of U2OS/pSUPER-c-FLIP-siRNA was further examined for TRAILinduced cell death and apoptosis in the presence of a pan-antagonist of inhibitor of apoptosis proteins (IAPs) AT406, with or without 4 hrs pretreatment with rocaglamide, an inhibitor of c-FLIP biosynthesis, for 24 hrs. Cell death effects and apoptosis were measured by the methods of MTT assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and flow cytometry, respectively. The results indicated that TRAIL-induced cell death in U2OS/pSUPER-c-FLIP-siRNA was increased compared with control cells U2OS/pSUPER in the presence or absence of AT406. Flow cytometry indicated that TRAIL-induced cell death effects proceeded through cell apoptosis pathway. However, in the presence of rocaglamide, cell death or apoptotic effects of TRAIL were similar and profound in both cell lines, suggesting that the mechanism of action for both c-FLIP-siRNA and rocaglamide was identical. We conclude that the inhibition of c-FLIP by either c-FLIP-siRNA or rocaglamide can enhance the sensitivity of U2OS to TRAIL-induced apopotosis, suggesting that inhibition of c-FLIP is a good target for anti-cancer therapy.
Dang, Yi-Wu;Zeng, Jing;He, Rong-Quan;Rong, Min-Hua;Luo, Dian-Zhong;Chen, Gang
Asian Pacific Journal of Cancer Prevention
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v.15
no.12
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pp.4969-4976
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2014
Background: miR-152 is involved in the genesis and development of several malignancies. However, its role in HCC has not been fully clarified. The aim of this study was to investigate the clinicopathological significance of miR-152 and its effect on the malignant phenotype of HCC cells. Methods: miR-152 expression was detected using real-time quantitative RT-PCR in 89 pairs of HCC formalin-fixed paraffin-embedded and their adjacent tissues. Functionally, in vitro effects and mechanisms of action of miR-152 on proliferation, viability, caspase activity, apoptosis and motility were explored in HepG2, HepB3 and SNU449 cells, as assessed by spectrophotometry, fluorimetry, fluorescence microscopy, wound-healing and Western blotting, respectively. Results: miR-152 expression in HCC was downregulated remarkably compared to that in adjacent hepatic tissues. miR-152 levels in groups of advanced clinical stage, larger tumor size and positive HBV infection, were significantly lower than in other groups. A miR-152 mimic could suppress cell growth, inhibit cell motility and increase caspase activity and apoptosis in HCC cell lines. Furthermore, Western blotting showed that the miR-152 mimic downregulated Wnt-1, DNMT1, ERK1/2, AKT and TNFRS6B signaling. Intriguingly, inverse correlation of TNFRF6B and miR-152 expression was found in HCC and bioinformatics confirmed that TNFRF6B might be a target of miR-152. Conclusions: Underexpression of miR-152 plays a vital role in hepatocarcinogenesis and lack of miR-152 is related to the progression of HCC through deregulation of cell proliferation, motility and apoptosis. miR-152 may act as a tumor suppressor miRNA by also targeting TNFRSF6B and is therefore a potential candidate biomarker for HCC diagnosis, prognosis and molecular therapy.
Lee, Ji Young;Jee, Su Bean;Park, Won Young;Choi, Yu Jin;Kim, Bokyung;Kim, Yoon Hee;Jun, Do Youn;Kim, Young Ho
Journal of Microbiology and Biotechnology
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v.24
no.12
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pp.1654-1663
/
2014
To examine the effect of tumor suppressor protein p53 on the antitumor activity of 2-methoxyestradiol (2-MeO-$E_2$), 2-MeO-$E_2$-induced cell cycle changes and apoptotic events were compared between the human colon carcinoma cell lines HCT116 ($p53^{+/+}$) and HCT116 ($p53^{-/-}$). When both cell types were exposed to 2-MeO-$E_2$, a reduction in the cell viability and an enhancement in the proportions of $G_2/M$ cells and apoptotic sub-$G_1$ cells commonly occurred dose-dependently. These 2-MeO-$E_2$-induced cellular changes, except for $G_2/M$ arrest, appeared to be more apparent in the presence of p53. Immunofluorescence microscopic analysis using anti-${\alpha}$-tubulin and anti-lamin B2 antibodies revealed that after 2-MeO-$E_2$ treatment, impaired mitotic spindle network and prometaphase arrest occurred similarly in both cell types. Following 2-MeO-$E_2$ treatment, only HCT116 ($p53^{+/+}$) cells exhibited an enhancement in the levels of p53, p-p53 (Ser-15), $p21^{WAF1/CIP1}$, and Bax; however, the Bak level remained relatively constant in both cell types, and the Bcl-2 level decreased only in HCT116 ($p53^{+/+}$) cells. Additionally, mitochondrial apoptotic events, including the activation of Bak and Bax, loss of ${\Delta}{\psi}m$, activation of caspase-9 and -3, and cleavage of lamin A/C, were more dominantly induced in the presence of p53. The Bak-specific and Bax-specific siRNA approaches confirmed the necessity of both Bak and Bax activations for the 2-MeO-$E_2$-induced apoptosis in HCT116 cells. These results show that among 2-MeO-$E_2$-induced apoptotic events, including prometaphase arrest, up-regulation of Bax level, down-regulation of Bcl-2 level, activation of both Bak and Bax, and mitochondria-dependent caspase activation, the modulation of Bax and Bcl-2 levels is the target of the pro-apoptotic action of p53.
Park, Sung-Jun;Hong, Sung-Ho;Lee, Anne Ha-Young;Kim, Cheol-Ju;Kim, Su-Jin;Kim, Sung-Kyoon;Ko, Gwang-Pyo
Journal of Environmental Health Sciences
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v.37
no.5
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pp.376-386
/
2011
Objectives: The aim of this study was to evaluate the microbial hazards posed by food utensils and fixtures in food service operations at selected middle and high schools located in Seoul, Korea. Methods: We collected 200 samples of utensils and fixtures including cups, spoons/chopsticks, food trays and tables from five different schools in Seoul. Target microorganisms of this study were divided into two groups: total bacterial count and total coliform as indicators of microbial contamination and Bacillus cereus and Staphylococcus aureus as pathogens of food poisoning. We used selective media to quantify microbial concentration and 16S rRNA PCR assay for qualitative analysis. In addition, intensive interviews with nutritionists were conducted and observations were made to identify factors that may affect microbial contamination. Logistic regression analysis was employed to examine the relationship between the microbial concentration and operation characteristics of each operation. Results: The level of microbial concentration in school B and C were significantly lower than in school A, D and E (p<0.05). Some samples from school A, D and E showed over 3.4 log CFU/100 $cm^2$ (total bacterial count) and 1.0 log CFU/100 $cm^2$ (total coliform), which requires immediate hygienic action. The number of customers per staff member, periodicity of hygiene education for staff and daily operation time of sterilizers were also found to be important factors related with the microbial contamination of food service operations. Conclusions: These results suggested that not only a HACCP (Hazard Analysis and Critical Control Point) approach, but also efforts to assess internal risk factors within operations be needed to reduce the microbial contamination of food utensils and fixtures. This study is expected to provide preliminary data for assessing microbial hazards in food service operations.
I. Introduction: In management control, business analysis has to do with a performance evaluation and is accounted much of manager's decision making, Business indicator accounting is the vehicle of decision making and also feedback can be accomplished by it. This study is to build up a logic about what capacity for use the business indicator accounting has in making decision. Therefore it is significant to make clear the adjustment of decision and to study the function of business indicator. II. Adjustment of Decision and Accounting Work: Adjustment of decision is connected with accounting now that business indicator accounting has a function of decision making. And it should be tied up with specialization as a structure of system, organization, communication and stage system. III. Adjustment of Decision and Managerial Accounting: Managerial account makes a great contributin to the management in which each adjustment of decision should be accomplished. Let me make mention of how the adjustment of decision is accomplished concretely, and what contribution che managerial accounting makes. In an adjustment of decision, centralization and decentrialization of enterprise are very import and I think the three problems, such as the extent of sphere (procurement, production and maketing), the face of affairs (planning, implementing and controlling), the final surge (decision making and action) can be accomplished by the business indicator accounting. IV. Structure of System and Disciplinary Approach for Decision: Decision can be classified into syncronized decision and continuous decision, and is closely connected with centralization and decentralization. In the course of systematizing, the sort of decision is classified into a man in charge of decision, and object of decision, conditions of decision, and an adjusting of decision. For it's object, it has an analogical thinking and an analytic subdivision about the target area. And it is premised on getting a scientific understanding. I think a disciplinary approach remains in solving these intricate problems. V. Conclusion: In this study I dealt with a specialization as a structure in management system and a theory that adjustment is a necessary process in decision making. For an adjustment of decision, exchanging informations and communication are necessary, and accounting is in charge of the process. And then the centralization and decentralization of decision should be connected in the way of adjustment of decision. In case of decentralization, the adjustment of decision is accomplished by the exchanging informations through feedback, and in case of centralization, by the all-round planning. And also I found that syncronized decision and decentralized decision are linked together. It is natural that the function of business indicator accounting is called for to render more services for it. Therefore, according to the extent of centralization and decentralization accounting to adjust the decision, can be various. Consequently, in relation to the structure of system. I think it is necessary to make a theoretical and empirical study of the business indicator accounting.
The purpose of this study was to analyse between foreign curriculum and our high school industrial technology subject-matter curriculum, to review trend and stream of curriculum revision, and purpose and content system of subject-matter. This study was conducted through reviewing literature; research reference, journal, book, and Web materials. in this study, comparative target country was restricted to Japan, U. S. A., U. K., and N. Z., Australia that administer to similar subject with our industrial technology subject-matter. The major finding of this study were as follows: 1. A similar subject-matter with our industrial technology subject0matter was Japan' 'foundation of industrial technology' and 'project research', U. S. A.' 'technology' and etc, U. K.' 'design and technology', and N. Z.' 'technology', 'New South Wales in Australia' design and technology'. 2. The result of analysis to purpose and strength of subject-matter, our' industrial technology subject-matter was oriented to knowledge, understanding and career search in industrial area. but, the other was emphasized technological problem solving by process-based method with thinking and action. 3. In the curriculum content, our country was treat to content area of a broad industrial world. on the other hand, Japan; relationship between human and technology, environment, process technology and product technology, project research. U. S. A.; technology content standards by knowledge, process and context, U. K., N. Z., and Australia were focused 'design process'. Based on above results, the recommendation can be established as follows: 1. A study on the implementation of industrial technology curriculum. 2. A study on the perception and need assessment of expert and stakeholder about purpose and content system. of industrial technology subject-matter.
Purpose : Infection with Shiga-like toxin (SLT)-producing Escherichia coli, an emerging human pathogen found particularly in young children under 5 years of age, causes a spectrum of illnesses with high morbidity and mortality, ranging from diarrhea to hemorrhagic colitis and hemolytic uremic syndrome. Host mediators play an important role in the pathogenesis of SLT-I toxicity. The experiments described here were designed to investigate the effect of SLT-I on TNF-${\alpha}$ production and to understand the effect of TNF-${\alpha}$ on GB3 expression. We also further examine the relationship between the Gb3 level and the differential susceptibility of cells to the cytotoxic action of SLT-I. Methods : The effect of purified SLT-1 from E. coli O157 : H7 (ATCC 43890) on tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) production in Raw264.7 cells was investigated. Many mediators regulate endothelial cell membrane expression of the glycolipid globotriaosyleramide (Gb3), which serves as the toxin receptor, suggesting that the host response to the toxin or other bacterial products may contribute to pathogenesis by regulating target cell sensitivity to the toxins. Therefore, the relationships between Gb3 expression and cytotoxicity against SLT-I on three types of cells were evaluated. Results : Detectable levels of TNF-${\alpha}$ were produced as early as six hours after induction and continued to increase during 48 hours by SLT-I. It was also found that Vero cells and dendritic cells (DC2.4 cells) expressed high levels of Gb3, 83% and 68%, respectively, and that Raw264.7 cells had a low level of Gb3 (29%) and appeared refractory to cytotoxicity against SLT-I. Vero cells and DC2.4 cells expressing high levels of Gb3 were highly susceptible to SLT-I. Furthermore, macrophages showed a resistance to SLT-I cytotoxicity, despite the fact that Gb3 expression was enhanced. Conclusion : These results strongly suggest that the expression of Gb3 is necessary but not sufficient to confer sensitivity of macrophages to SLT-I and further underpin the important role of SLT-I and its Gb3 receptors in the pathogenesis of E. coli O157 infection.
Park, Ju Ha;Guo, Lu;Kang, He Mi;Son, Beung Gu;Kang, Jum Soon;Lee, Yong Jae;Park, Young Hoon;Je, Byoung Il;Choi, Young Whan
Journal of Life Science
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v.31
no.2
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pp.209-218
/
2021
This study aimed to evaluate the anti-obesity effects of Cudrania tricuspidata leaf extract in the order of leaf development on the shoot (L0, L1, L2, L3, L4, and L5). The leaves at the apex of a Cudrania tricuspidata shoot were classified as L0; the next leaves of the apex were classified as L1, L2, L3, and L4 from highest to lowest; and the lowest leaf was classified as L5. A series of 70% ethyl alcohol leaf extracts were screened for the inhibitory effects of adipogenesis in 3T3-L1 preadipocytes. We found that the apical leaf extract of Cudrania tricuspidata (CTL0) was the most effective. Next, a study was conducted on the inhibitory action mechanism of CTL0. Treatment with CTL0 significantly suppressed the differentiation of 3T3-L1 preadipocytes in a dose-dependent manner, as confirmed by the decrease in lipid droplet content observed with Oil Red O staining. Treatment with 12.5 ㎍/ml, 25 ㎍/ml, and 50 ㎍/ml of CTL0 significantly reduced the lipid droplet content. Glucose and cellular triglyceride concentrations were reduced in the 3T3-L1 cells on the CTL0-treated medium compared to the differentiation medium (DM control, DMEM + insulin + dexamethasone + rosiglitazone). Compared with DM, CTL0 significantly inhibited the expression of key pro-adipogenic transcription factors, including peroxisome proliferator-activated receptor γ (PPARγ), LPL, A-FABP, and Glut4. These findings show that CTL0 extract has potent anti-obesity effects.
This study aims to explore the direction for Korea's effective response to Target 3 (30by30), which can be said to be the core of the Kunming-Montreal Global Biodiversity Framework (K-M GBF) of the Convention on Biological Diversity (CBD), to find the direction of systematic OECM (Other Effective area-based Conservation Measures) discovery at the national level through a survey of global conceptual review and expert perception of OECM. This study examined ① the use of Korean terms related to OECM, ② derivation of determining criteria reflecting global standards, ③ deriving types of potential OECM candidates in Korea, and ④ considerations for OECM identification and reporting to explore the direction for identifying systematic, national-level OECM that complies with global standards and reflects the Korean context. First, there was consensus for using Korean terminology that reflects the concept of OECM rather than simple translations, and it was determined that "nature coexistence area" was the most preferred term (12 people) and had the same context as CBD 2050 Vision of "a world of living in harmony with nature." This study suggests utilizing four criteria (1. No protected areas, 2. Geographic boundaries, 3. Governance/management, and 4. Biodiversity value) that reflect OECM's core characteristics in the first-stage selection process, carrying out the consensus-building process (stage 2) with the relevant agencies, and adding two criteria (3-1 Effectiveness and sustainability of governance and management and 4-1 Long-term conservation) and performing the in-depth diagnosis in stage 3 (full assessment for reporting). The 28 types examined in this study were generally compatible with OECMs (4.45-6.21/7 points, mean 5.24). In particular, the "Conservation Properties (6.21 points)" and "Conservation Agreements (6.07 points)", which are controlled by National Nature Trust, are shown to be the most in line with the OECM concept. They were followed by "Buffer zone of World Natural Heritage (5.77 points)", "Temple Forest (5.73 points)", "Green-belt (Restricted development zones, 5.63 points)", "DMZ (5.60 points)", and "Buffer zone of biosphere reserve (5.50 point)" to have high potential. In the case of "Uninhabited Islands under Absolute Conservation", the response that they conformed to the protected areas (5.83/7 points) was higher than the OECM compatibility (5.52/7 points), it is determined that in the future, it would be preferable to promote the listing of absolute unprotected islands in the Korea Database on Protected Areas (KDPA) along with their surrounding waters (1 km). Based on the results of a global OECM standard review and expert perception survey, 10 items were suggested as considerations when identifying OECM in the Korean context. In the future, continuous research is needed to identify the potential OECMs through site-level assessment regarding these considerations and establish an effective in-situ conservation system at the national level by linking existing protected area systems and identified OECMs.
The naturally occurring chemical indole-3-carbinol (13C), found in vegetables of the Brassica genus, is a promising anticancer agent that was shown previ- ously to induce a Gl cell cycle arrest of human breast cancer cell lines, independent of estrogen receptor signaling. The anticancer activity of 13C and the possible mechanisms of its action were explored in a human hepatocellular carcinoma cell line, HepG2. Treatment of HepG2 cells with 13C suppressed the growth of the cells. The growth sup- pression caused by 13C ($IC_{50}$/: 444$\mu$M) was found to be partially due to its ability to stop the cell cycle in HepG2 cells. Western blot analysis for the Gl phase artiest demonstrated that the expression-levels of cyclin-dependent kinase (Cdk4, Cdk6) and cyclic D were reduced strongly after treatment of Hep72 cells with 13C (4007M) for 24- 72 hrs. Furthermore, I3C selectively abolished the expression of Cdk6 in a dose- and time-dependent manner, and accordingly, inhibited the phosphorylation of retinoblastoma. Interestingly, after the HepG2 cells reached their max- imal growth arrest, the level of the p21, a well-known Cdk inhibitor, increased significantly. Therefore, it could be considered that the Gl arrest of HepG2 cells treated with 13C was due to the indirect inhibition of Cdk4/6 activities by p21 Western blot analysis for G2/M phase arrest of demonstrated the levels of Cdc2 and cyclin Bl werer reduced dramatically after the treatment of HepG2 cells with 13C ($40\mu$M) for 24-72 hrs. flow cytometry of propidium iodide-stained HepG2 cells revealed that 13C induces a Gl (53%,72hr incubation) and G2 (25%,24hr incubation) cell cycle arrest. Thus, our observations have uncovered a previously undefined antiproliferative pathway for r3C that implicates Cdk4/6 and Cdc2 as a target for cell cycle control in human HepG2 cells. However, the 13C-medi- ated cell cycle arrest and repression of Cdk4/6 production did not affect the apoptotic induction of HepG2 cell.
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