• BMB Reports
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• v.44 no.7
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• pp.490-495
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• 2011

Transcription factor AP-$2{\alpha}$ involves in the process of mammalian embryonic development and tumorigenesis. Many studies have shown that AP-$2{\alpha}$ functions in association with other interacting proteins. In a two-hybrid screening, the regulatory subunit ${\beta}$ of protein casein kinase 2 ($CK2{\beta}$) was identified as an interacting protein of AP-$2{\alpha}$; we confirmed this interaction using in-vitro GST pull-down and in-vivo co-immunoprecipitation assays; in an endogenous co-immunoprecipitation experiment, we further found the catalytic subunit ${\alpha}$ of protein casein kinase 2 ($CK2{\alpha}$) also exists in the complex. Phosphorylation analysis revealed that AP-$2{\alpha}$ was phosphorylated by CK2 kinase majorly at the site of Ser429, and such phosphorylation could be blocked by CK2 specific inhibitor 4,5,6,7-tetrabromobenzotriazole (TBB) in a dose-dependent manner. Luciferase assays demonstrated that both $CK2{\alpha}$ and $CK2{\beta}$ enhanced the transcription activity of AP-$2{\alpha}$; moreover, $CK2{\beta}$ increased the stability of AP-$2{\alpha}$. Our data suggest a novel cellular function of CK-2 as a transcriptional co-activator of AP-$2{\alpha}$.

• BMB Reports
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• v.43 no.6
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• pp.432-437
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• 2010

LBH is a transcription factor as a candidate gene for CHD associated with partial trisomy 2p syndrome. To identify potential LBH-interacting partners, a yeast two-hybrid screen using LBH as a bait was performed with a human heart cDNA library. One of the clones identified encodes ${\alpha}B$-crystallin. Co-immunoprecipitation and GST pull-down assays showed that LBH interacts with ${\alpha}B$-crystallin, which is further confirmed by mammalian two-hybrid assays. Co-localization analysis showed that in COS-7 cells, ${\alpha}B$-crystallin that is cytoplasmic alone, accumulates partialy in the nucleus when co-transfected with LBH. Transient transfection assays indicated that overexpression of LBH or ${\alpha}B$-crystallin reduced the transcriptional activities of p53 and p21, respectively, Overexpression of both ${\alpha}B$-crystallin and LBH together resulted in a stronger repression of the transcriptional activities of p21 and p53. These results showed that the interaction of LBH and ${\alpha}B$-crystallin may inhibit synergistically the transcriptional regulation of p53 and p21.

• Molecules and Cells
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• v.38 no.3
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• pp.259-266
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• 2015

The regulation of flowering time has crucial implications for plant fitness. MicroRNA156 (miR156) represses the floral transition in Arabidopsis thaliana, but the mechanisms regulating its transcription remain unclear. Here, we show that two AGAMOUS-like proteins, AGL15 and AGL18, act as positive regulators of the expression of MIR156. Small RNA northern blot analysis revealed a significant decrease in the levels of mature miR156 in agl15 agl18 double mutants, but not in the single mutants, suggesting that AGL15 and AGL18 co-regulate miR156 expression. Histochemical analysis further indicated that the double mutants showed a reduction in MIR156 promoter strength. The double mutants also showed reduced abundance of pri-miR156a and pri-miR156c, two of the primary transcripts from MIR156 genes. Electrophoretic mobility shift assays demonstrated that AGL15 directly associated with the CArG motifs in the MIR156a/c promoters. AGL18 did not show binding affinity to the CArG motifs, but pull-down and yeast two-hybrid assays showed that AGL18 forms a heterodimer with AGL15. GFP reporter assays and bimolecular fluorescence complementation (BiFC) showed that AGL15 and AGL18 co-localize in the nucleus and confirmed their in vivo interaction. Overexpression of miR156 did not affect the levels of AGL15 and AGL18 transcripts. Taking these data together, we present a model for the transcriptional regulation of MIR156. In this model, AGL15 and AGL18 may form a complex along with other proteins, and bind to the CArG motifs of the promoters of MIR156 to activate the MIR156 expression.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.9
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• pp.3723-3727
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• 2015

The p53 tumor suppressor protein is a principal mediator of growth arrest, senescence, and apoptosis in response to a broad array of cellular damage. p53 is a substrate for the ubiquitin-proteasome system, however, the ubiquitin-conjugating enzymes (E2s) involved in p53 ubiquitination have not been well studied. UBE2Q1 is a novel E2 ubiquitin conjugating enzyme gene. Here, we investigated the effect of UBE2Q1 overexpression on the level of p53 in the MDA-MB-468 breast cancer cell line as well as the interaction between UBE2Q1 and p53. By using a lipofection method, the p53 mutated breast cancer cell line, MDA-MB-468, was transfected with the vector pCMV6-AN-GFP, containing UBE2Q1 ORF. Western blot analysis was employed to verify the overexpression of UBE2Q1 in MDA-MB-468 cells and to evaluate the expression level of p53 before and after cell transfection. Immunoprecipitation and GST pull-down protocols were used to investigate the binding of UBE2Q1 to p53. We established MDA-MB-468 cells that transiently expressed a GFP fusion proteins containing UBE2Q1 (GFP-UBE2Q1). Western blot analysis revealed that levels of p53 were markedly lower in UBE2Q1 transfected MDA-MB-468 cells as compared with control MDA-MB-468 cells. Both in vivo and in vitro data showed that UBE2Q1 co-precipitated with p53 protein. Our data for the first time showed that overexpression of UBE2Q1can lead to the repression of p53 in MDA-MB-468 cells. This repression of p53 may be due to its UBE2Q1 mediated ubiquitination and subsequent proteasome degradation, a process that may involve direct interaction of UBE2Q1with p53.

• Journal of Korean Tunnelling and Underground Space Association
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• v.19 no.4
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• pp.651-668
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• 2017

For initial stability of the tunnel, the primary support, Shotcrete and rockbolt shall be placed in the most appropriate time. This is because the role of such support plays a vital role in long-term and short-term tunnel stability. In this study, the rock bolt is an important supporting system that receives the external pressure generated by the stress relaxation during tunnel excavation as axial force and transmits it to the shotcrete on the tunnel excavation surface. Until now, most of the materials of rock bolts have been used in the field, but there have been many problems such as uncertain quality of Chinese materials entering the market, poor packing due to falling down of rock bolts when filled with mortar, and corrosion due to water. Therefore, in this study, we have developed a high strength steel pipe rock bolt using Autobeam material to solve and improve various problems of existing rock bolts. In order to evaluate the performance of the developed bolt, field tests were carried out and the existing mortar filler in order to improve the performance of the rock bolt, the design and construction criteria were studied and the results were included in this paper.

• Journal of Pharmacopuncture
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• v.13 no.2
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• pp.93-100
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• 2010

Background: Auricular acupressure is one of the traditional health care treatments in oriental medicine. Approximately, 30~40% of the cancer patients have said to be suffering from insomnia and half of them having chronic and severe insomnia at the same time. Insomnia caused cancer patients feel more pain, fatigue, depression and anxiety and it sometimes let the power to have the best of cancer pull down. Objective: To investigate how effective the auricular acupressure treatment to cancer patients suffering from insomnia. Methods: We recruited participants from East-West Cancer Center of Daejeon University. Finally, of the people whose age range from 20 to 75, 12 patients who got less than 40 points from the score of Oh's sleeping score (OSS) were recruited. Single-blind, randomized pilot study was performed. The treatment group received auricular acupressure treatment (AAT) on active points and the control group had received sham acupressure treatment (SAT) for five times. Sleep parameters were checked by using OSS and numeric rating scale (NRS). We checked the scale everytime, both before and after treatment. We analyzed the data statistically by using independent T-test, paired T-test and analysis of variance (ANOVA) test. (p<0.05) Results: Twelve cancer patients participated in this pilot study and there was no significant difference between control and treatment group. Only 7 of them had completed the whole treatment process, 4 patients of AAT group and 3 participants of SAT. The OSS of AAT group had increased from $34.0{\pm}4.3$ to $39.5{\pm}3.1$ and that of SAT group had increased from $38.3{\pm}3.5$ to $40.0{\pm}0.0$. There was no significant difference between them. The NRS of AAT group had increased from $6.3{\pm}2.9$, $04.8{\pm}2.1$ and that of SAT group had increased from $7.0{\pm}1.0$ to $5.0{\pm}2.6$. No significant difference was observed between them. Conclusion: Although both groups did not show significant differences, most of the experimental participants showed increasing OSS and NRS after treatments. Significant participants' number will be needed in the next study.

• Korean Journal of Materials Research
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• v.3 no.2
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• pp.121-130
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• 1993

The GaP crystals were grown by synthesis solute diffusion method and its properties were investigated. High quality single crystals were obtained by pull-down the crystal growing ampoule with velocity of 1.75mm/day. Etch pits density along vertical direction of ingot was increased from 3.8 ${\times}{10^4}$c$m^{-2}$ of the first freeze to 2.3 ${\times}{10^5}$c$m^2$ of the last freeze part. The carrier concentration and mobilities at room temperature were measured to 197.49cc$m^2$/V.sec and 6.75 ${\times}{10^{15}}$c$m^{-3]$, respectively. The temperature dependence of optical energy gap was empirically fitted to $E_g$(T)=[2.3383-(6.082${\times}{10^{-4}}$)$T^2$/(373. 096+TJeV. Photoluminescence spectra measured at low temperature were consist with sharp line-spectra near band-gap energy due to bound-exciton and phonon participation in band edge recombination process. Zn-diffusion depth in GaP was increased with square root of diffusion time and temperature dependence of diffusion coefficient was D(Tl = 3.2 ${\times}{10^3}$exp( - 3.486/$k_{\theta}$T)c$m^2$/sec. Electroluminescence spectra of p-n GaP homojunction diode are consisted with emission at 630nm due to recombination of donor in Zn-O complex center with shallow acceptors and near band edge emission at 550nm. Photon emission at current injection level of lower than 100m A was due to the band-filling mechanism.

• The Korean Journal of Physiology and Pharmacology
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• v.26 no.4
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• pp.239-253
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• 2022

Epithelial-mesenchymal transition (EMT) is known to be involved in airway remodeling and fibrosis of bronchial asthma. However, the molecular mechanisms leading to EMT have yet to be fully clarified. The current study was designed to reveal the potential mechanism of microRNA-21 (miR-21) and poly (ADP-ribose) polymerase-1 (PARP-1) affecting EMT through the PI3K/AKT signaling pathway. Human bronchial epithelial cells (16HBE cells) were transfected with miR-21 mimics/inhibitors and PARP-1 plasmid/small interfering RNA (siRNA). A dual luciferase reporter assay and biotin-labeled RNA pull-down experiments were conducted to verify the targeting relationship between miR-21 mimics and PARP-1. The migration ability of 16HBE cells was evaluated by Transwell assay. Quantitative real-time polymerase chain reaction and Western blotting experiments were applied to determine the expression of Snail, ZEB1, E-cadherin, N-cadherin, Vimentin, and PARP-1. The effects of the PI3K inhibitor LY294002 on the migration of 16HBE cells and EMT were investigated. Overexpression of miR-21 mimics induced migration and EMT of 16HBE cells, which was significantly inhibited by overexpression of PARP-1. Our findings showed that PARP-1 was a direct target of miR-21, and that miR-21 targeted PARP-1 to promote migration and EMT of 16HBE cells through the PI3K/AKT signaling pathway. Using LY294002 to block PI3K/AKT signaling pathway resulted in a significant reduction in the migration and EMT of 16HBE cells. These results suggest that miR-21 promotes EMT and migration of HBE cells by targeting PARP-1. Additionally, the PI3K/AKT signaling pathway might be involved in this mechanism, which could indicate its usefulness as a therapeutic target for asthma.

• Journal of Ginseng Research
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• v.46 no.3
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• pp.396-407
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• 2022

Background: Colorectal cancer (CRC) has a high morbidity and mortality worldwide. 20 (S)-ginsenoside Rh2 (G-Rh2) is a natural compound extracted from ginseng, which exhibits anticancer effects in many cancer types. In this study, we demonstrated the effect and underlying molecular mechanism of G-Rh2 in CRC cells in vitro and in vivo. Methods: Cell proliferation, migration, invasion, apoptosis, cell cycle, and western blot assays were performed to evaluate the effect of G-Rh2 on CRC cells. In vitro pull-down assay was used to verify the interaction between G-Rh2 and Axl. Transfection and infection experiments were used to explore the function of Axl in CRC cells. CRC xenograft models were used to further investigate the effect of Axl knockdown and G-Rh2 on tumor growth in vivo. Results: G-Rh2 significantly inhibited proliferation, migration, and invasion, and induced apoptosis and G₀/G₁ phase cell cycle arrest in CRC cell lines. G-Rh2 directly binds to Axl and inhibits the Axl signaling pathway in CRC cells. Knockdown of Axl suppressed the growth, migration and invasion ability of CRC cells in vitro and xenograft tumor growth in vivo, whereas overexpression of Axl promoted the growth, migration, and invasion ability of CRC cells. Moreover, G-Rh2 significantly suppressed CRC xenograft tumor growth by inhibiting Axl signaling with no obvious toxicity to nude mice. Conclusion: Our results indicate that G-Rh2 exerts anticancer activity in vitro and in vivo by suppressing the Axl signaling pathway. G-Rh2 is a promising candidate for CRC prevention and treatment.

• Asia-Pacific Journal of Business Venturing and Entrepreneurship
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• v.18 no.2
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• pp.67-79
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• 2023

The bounce rate is the rate at which a user leaves immediately after visiting, and this study aimed to find out what attributes of a website affect the bounce rate. Web site evaluation items were defined as a total of 4 items and 27 evaluation attributes, including usability, information, service interaction, and technology, so that they can be commonly applied to venture companies in various industries through prior research. As a result of the study, 6 website attributes that affect the bounce rate were verified to be significant by discriminant analysis and decision tree analysis. Suggestions to reduce the bounce rate of venture business websites through this study are as follows. First, the path name of the website is displayed as mandatory and a pull-down menu function is added to facilitate movement to other pages. Second, it is good to expose core content that can attract users' attention in the form of a banner, and place internal link banners in the right place on sub-pages. Third, external links should be linked to a new window so that they do not leave the current page immediately so that they can be re-entered. Lastly, it is recommended to expose the contact information of the person in charge and consultation function as direct information for communication with customers, but if individual response is difficult, at least the consultation function must be added. These suggestions are expected to be of practical help in various fields such as website development, operation, and marketing. However, in special cases, a high bounce rate may be normal, so it should be considered according to the situation.