Gα₁₂ gep oncogene deregulation of p53-responsive microRNAs promotes epithelial–mesenchymal transition of hepatocellular carcinoma

Yang, Y M,Lee, W H,Lee, C G,An, J,Kim, E-S,Kim, S H,Lee, S-K,Lee, C H,Dhanasekaran, D N,Moon, A,Hwang, S,Lee, S J,Park, J-W,Kim, K M,Kim, S G Macmillan Publishers Limited 2015 Oncogene Vol.34 No.22

Hepatocellular carcinoma (HCC) has a poor prognosis owing to aggressive phenotype. Gα<SUB>12</SUB> gep oncogene product couples to G-protein-coupled receptors, whose ligand levels are frequently increased in tumor microenvironments. Here, we report Gα<SUB>12</SUB> overexpression in human HCC and the resultant induction of zinc-finger E-box-binding homeobox 1 (ZEB1) as mediated by microRNA deregulation. Gα<SUB>12</SUB> expression was higher in HCC than surrounding non-tumorous tissue. Transfection of Huh7 cell with an activated mutant of Gα<SUB>12</SUB> (Gα<SUB>12</SUB>QL) deregulated microRNA (miRNA or miR)-200b/a/429, -194-2/192 and -194-1/215 clusters in the miRNome. cDNA microarray analyses disclosed the targets affected by Gα<SUB>12</SUB> gene knockout. An integrative network of miRNAs and mRNA changes enabled us to predict ZEB1 as a key molecule governed by Gα<SUB>12</SUB>. Decreases of miR-200a/b, -192 and -215 by Gα<SUB>12</SUB> caused ZEB1 induction. The ability of Gα<SUB>12</SUB> to decrease p53 levels, as a result of activating protein-1 (AP-1)/c-Jun-mediated mouse double minute 2 homolog induction, contributed to transcriptional deregulation of the miRNAs. Gα<SUB>12</SUB>QL induced ZEB1 and other epithelial–mesenchymal transition markers with fibroblastoid phenotype change. Consistently, transfection with miR-200b, -192 or -215 mimic prevented the ability of Gα<SUB>12</SUB>QL to increase tumor cell migration/invasion. In xenograft studies, sustained knockdown of Gα<SUB>12</SUB> decreased the overall growth rate and average volume of tumors derived from SK-Hep1 cell (mesenchymal-typed). In HCC patients, miR-192, -215 and/or -200a were deregulated with microvascular invasion or growth advantage. In the HCC samples with higher Gα<SUB>12</SUB> level, a correlation existed in the comparison of relative changes of Gα<SUB>12</SUB> and ZEB1. In conclusion, Gα<SUB>12</SUB> overexpressed in HCC causes ZEB1 induction by deregulating p53-responsive miRNAs, which may facilitate epithelial–mesenchymal transition and growth of liver tumor. These findings highlight the significance of Gα<SUB>12</SUB> upregulation in liver tumor progression, implicating Gα<SUB>12</SUB> as an attractive therapeutic target.

Purification and characterization of a novel fibrinolytic α chymotrypsin like serine metalloprotease from the edible mushroom, Lyophyllum shimeji

Moon, S.M.,Kim, J.S.,Kim, H.J.,Choi, M.S.,Park, B.R.,Kim, S.G.,Ahn, H.,Chun, H.S.,Shin, Y.K.,Kim, J.J.,Kim, D.K.,Lee, S.Y.,Seo, Y.W.,Kim, Y.H.,Kim, C.S. Society for Bioscience and Bioengineering, Japan ; 2014 Journal of bioscience and bioengineering Vol.117 No.5

A novel fibrinolytic enzyme was purified from Lyophyllum shimeji, a popular edible mushroom in Asia. The enzyme was purified using combination of anion exchange chromatography on a Mono Q 5/5 column and size exclusion gel filtration chromatography on Superdex 200 100/300 column. This purification protocol resulted 80.9-fold purification of the enzyme and a final yield of 5.7%. The molecular weight of the purified enzyme was estimated to be 21 kDa by SDS-PAGE and size exclusion gel filtration. The N-terminal amino acid sequence was found to be ITFQSASP, which is dissimilar from that of known fibrinolytic enzymes. The purified enzyme was a neutral protease with an optimal reaction pH and temperature of 8.0 and 37<SUP>o</SUP>C, respectively. Enzymatic activity was inhibited by Cu<SUP>2+</SUP> and Co<SUP>2+</SUP>. It was also significantly inhibited by PMSF and TPCK. Furthermore, it was found to exhibit a higher specificity for S-7388, a well-known chymotrypsin chromogenic substrate, indicating chymotrypsin like serine metalloprotease. The relative fibrinolytic activity of 5 μg purified enzyme have two fold more activity than 1 unit/ml of plasmin on fibrin plate. Furthermore, purified enzyme preferentially hydrolyzed the Aα-chain followed by the Bβ- and γ-chain of fibrinogen, which is precursor of fibrin. Therefore, these data suggests that the fibrinolytic enzyme derived from edible mushroom, L. shimeji, might be useful for thrombolytic therapy and preventing thrombotic disease.

Species and gamete-specific fertilization success of two sea urchins under near future levels of pCO₂

Sung, C.G.,Kim, T.W.,Park, Y.G.,Kang, S.G.,Inaba, K.,Shiba, K.,Choi, T.S.,Moon, S.D.,Litvin, S.,Lee, K.T.,Lee, J.S. Elsevier 2014 Journal of marine systems Vol.137 No.-

Since the Industrial Revolution, rising atmospheric CO<SUB>2</SUB> concentration has driven an increase in the partial pressure of CO<SUB>2</SUB> in seawater (pCO<SUB>2</SUB>), thus lowering ocean pH. We examined the separate effects of exposure of gametes to elevated pCO<SUB>2</SUB> and low pH on fertilization success of the sea urchin Strongylocentrotus nudus. Sperm and eggs were independently exposed to seawater with pCO<SUB>2</SUB> levels ranging from 380 (pH7.96-8.3) to 6000ppmv (pH7.15-7.20). When sperm were exposed, fertilization rate decreased drastically with increased pCO<SUB>2</SUB>, even at a concentration of 450ppmv (pH range: 7.94 to 7.96). Conversely, fertilization of Hemicentrotus pulcherrimus was not significantly changed even when sperm was exposed to pCO2 concentrations as high as 750ppmv. Exposure of S. nudus eggs to seawater with high pCO<SUB>2</SUB> did not affect fertilization success, suggesting that the effect of increased pCO<SUB>2</SUB> on sperm is responsible for reduced fertilization success. Surprisingly, this result was not related to sperm motility, which was insensitive to pCO<SUB>2</SUB>. When seawater was acidified using HCl, leaving pCO<SUB>2</SUB> constant, fertilization success in S. nudus remained high (>80%) until pH decreased to 7.3. While further studies are required to elucidate the physiological mechanism by which elevated pCO<SUB>2</SUB> impairs sperm and reduces S. nudus fertilization, this study suggests that in the foreseeable future, sea urchin survival may be threatened due to lower fertilization success driven by elevated pCO<SUB>2</SUB> rather than by decreased pH in seawater.

Inflammatory lipid sphingosine-1-phosphate upregulates C-reactive protein via C/EBPβ and potentiates breast cancer progression

Kim, E-S,Cha, Y,Ham, M,Jung, J,Kim, S G,Hwang, S,Kleemann, R,Moon, A Macmillan Publishers Limited 2014 Oncogene Vol.33 No.27

A crucial role of the inflammatory lipid sphingosine-1-phosphate (S1P) in breast cancer aggressiveness has been reported. Recent clinical studies have suggested that C-reactive protein (CRP) has a role in breast cancer development. However, limited information is available on the molecular basis for the expression of CRP and its functional significance in breast cell invasion. The present study aimed to elucidate the molecular link between S1P and CRP during the invasive process of breast epithelial cells. This is the first report showing that transcription of CRP was markedly activated by S1P in breast cells. Our data suggest that not only S1P treatment but also the endogenously produced S1P may upregulate CRP in breast carcinoma cells. Transcription factors CCAAT/enhancer-binding protein beta and c-fos were required for S1P-induced CRP expression. Coupling of S1P<SUB>3</SUB> to heterotrimeric G<SUB>αq</SUB> triggered the expression of CRP, utilizing signaling pathways involving reactive oxygen species (ROS), Ca<SUP>2+</SUP> and extracellular signal-related kinases (ERKs). S1P-induced CRP expression was crucial for the transcriptional activation of matrix metalloproteinase-9 through ERKs, ROS and c-fos, leading to breast cell invasion. Using a xenograft mice tumor model, we demonstrated that S1P induced CRP expression both in vitro and in vivo. Taken together, our findings have revealed a molecular basis for S1P-induced transcriptional activation of CRP and its functional significance in the acquisition of the invasive phenotype of human breast epithelial cells under inflammatory conditions. Our findings may provide useful information on the identification of useful therapeutic targets for inflammatory breast cancer.

Quality characteristics of common wheat fresh noodle with insoluble dietary fiber from kimchi by-product

Kim, B.R.,Kim, S.,Bae, G.S.,Chang, M.B.,Moon, B. ACADEMIC PRESS - JOURNALS DEPARTMENT 2017 FOOD SCIENCE AND TECHNOLOGY -ZURICH- Vol.85 No.PA

<P>This study aimed to investigate the quality of fresh noodles prepared using insoluble dietary fiber-enriched fractions (IEF) recovered from kimchi by-products (KBP) and common wheat flour, instead of semolina. Four samples (NICO, NIC2, NIC4, and NIC6) were prepared using common wheat flour mixed with different ratios of IEF (0 g/100 g; 2 g/100 g; 4 g/100 g; 6 g/100 g, respectively); Semolina pasta (SMP) was prepared as control. With increasing IEF ratios, cooking loss and swelling indexes increased significantly (p < 0.05). SEM images showed continuous and clear air pockets that were observed in NIC2 and SMP, whereas a loose protein network and big air pockets were observed in NIC4 and NIC6. After addition of IEF, L*, a*, and b* of samples decreased in all samples. Hardness of all samples in TPA was not statistically different regardless of the addition of IEF to common wheat flour. NIC2 showed the highest overall acceptability in sensory evaluation, while hardness, texture, color, taste, and flavor decreased in higher than 2 g/100 g (p < 0.05) IEF. Based on these results, fresh noodle using 2 g/100 g IEF from KBP and common wheat flour showed a great possibility to replace semolina pasta. (C) 2017 Elsevier Ltd. All rights reserved.</P>

문학반응이론에 의한 보건의학기술계열 국가시험개선방안에 관한 연구

이창규,이승관,조경진,박종성,정수경,유병서,박상숙,윤효숙,황선철,문경환,김정민,함용운,김지환,임국환,김영순,윤경희,황성준 高麗大學校 倂設 保健大學 保健科學硏究所 1998 保健科學論集 Vol.24 No.1

The nationally-governed examinations for certification of allied health professions in Korea have been continued for thirty three years. During that time, there were a lot of managerial improvements in carrying out the examinations, for example, the looking-over the papers converted from manual method to computerized one. Nevertheless, the overall aspects of item management in the national examinations are still remained as obsolete style. In some developed countries they have already tried or adopted computerized system in making questions, executing item analysis, developing item banking, and in overall management of examinations, looking over papers for their national licensure examinations, and have established good reputations. Since the National Health Personnel Licensing Examination Board was established in 1998, now we can expect there would be a lots of improvements in the managerial systems and organizational structures associated with the national licensing examinations suggesting followings. 1. The contents and scope of the licensing examinations based on job analysis should be announced publicaly. 2. Items should be developed based on the scopes and contents of job characteristics. 3. All the developed items for the licensing examinations should be tested quantitatively prior to banking items. 4. All the parameters of the stored items should be fully estimated through item response theory. 5. All the efforts should be given in order to execute the examinations in individual residential areas on behalf of every examinee. 6. To the licensing examination system the Computerized Adaptive Testing system should be introduced in order to enhance the efficiencies. 7. Security enhancement on the stored items should be reminded in order to prevent leaking out the banking items. 8. Much more improvements should be made for the proper job evaluations. 9. Every efforts should be given to prevent cheatings encountable during the examinations. 10. Make the best of professional volunteers from the various fields in the execution of licensing examinations. 11. Consider a new system in that examinations can be executed twice or more in a year. 12. The current methods of presiding over the examinations should be replaced by a more reasonable one. 13. The results of the examinations should be announced as soon as possible in order not to prolong the examinees' unemployment periods. 14. The National Health Personnel Licensing Examination Board should try to rationalize the management keeping step with the information-oriented society.

The E3 ubiquitin ligase CHIP selectively regulates mutant epidermal growth factor receptor by ubiquitination and degradation

Chung, C.,Yoo, G.,Kim, T.,Lee, D.,Lee, C.S.,Cha, H.R.,Park, Y.H.,Moon, J.Y.,Jung, S.S.,Kim, J.O.,Lee, J.C.,Kim, S.Y.,Park, H.S.,Park, M.,Park, D.I.,Lim, D.S.,Jang, K.W.,Lee, J.E. Academic Press 2016 Biochemical and biophysical research communication Vol.479 No.2

Somatic mutation in the tyrosine kinase domain of epidermal growth factor receptor (EGFR) is a decisive factor for the therapeutic response to EGFR tyrosine kinase inhibitors (EGFR-TKIs) in lung adenocarcinoma. The stability of mutant EGFR is maintained by various regulators, including heat shock protein 90 (Hsp90). The C terminus of Hsc70-interacting protein (CHIP) is a Hsp70/Hsp90 co-chaperone and exhibits E3 ubiquitin ligase activity. The high-affinity Hsp90-CHIP complex recognizes and selectively regulates their client proteins. CHIP also works with its own E3 ligase activity independently of Hsp70/Hsp90. Here, we investigated the role of CHIP in regulating EGFR in lung adenocarcinoma and also evaluated the specificity of CHIP's effects on mutant EGFR. In HEK 293T cells transfected with either WT EGFR or EGFR mutants, the overexpression of CHIP selectively decreased the expression of certain EGFR mutants (G719S, L747_E749del A750P and L858R) but not WT EGFR. In a pull-down assay, CHIP selectively interacted with EGFR mutants and simultaneously induced their ubiquitination and proteasomal degradation. The expressions of mutant EGFR in PC9 and H1975 were diminished by CHIP, while the expression of WT EGFR in A549 was nearly not affected. In addition, CHIP overexpression inhibited cell proliferation and xenograft's tumor growth of EGFR mutant cell lines, but not WT EGFR cell lines. EGFR mutant specific ubiquitination by CHIP may provide a crucial regulating mechanism for EGFR in lung adenocarcinoma. Our results suggest that CHIP can be novel therapeutic target for overcoming the EGFR TKI resistance.

Inhibitory effects of sulfur compounds on methane oxidation by a methane-oxidizing consortium

Lee, E.H.,Moon, K.E.,Kim, T.G.,Lee, S.D.,Cho, K.S. Society for Bioscience and Bioengineering, Japan ; 2015 Journal of bioscience and bioengineering Vol.120 No.6

Kinetic and enzymatic inhibition experiments were performed to investigate the effects of methanethiol (MT) and hydrogen sulfide (H<SUB>2</SUB>S) on methane oxidation by a methane-oxidizing consortium. In the coexistence of MT and H<SUB>2</SUB>S, the oxidation of methane was delayed until MT and H<SUB>2</SUB>S were completely degraded. MT and H<SUB>2</SUB>S could be degraded, both with and without methane. The kinetic analysis revealed that the methane-oxidizing consortium showed a maximum methane oxidation rate (V<SUB>max</SUB>) of 3.7 mmol g-dry cell weight (DCW)<SUP>-1</SUP> h<SUP>-1</SUP> and a saturation constant (K<SUB>m</SUB>) of 184.1 μM. MT and H<SUB>2</SUB>S show competitive inhibition on methane oxidation, with inhibition values (K<SUB>i</SUB>) of 1504.8 and 359.8 μM, respectively. MT was primary removed by particulate methane monooxygenases (pMMO) of the consortium, while H<SUB>2</SUB>S was degraded by the other microorganisms or enzymes in the consortium. DNA and mRNA transcript levels of the pmoA gene expressions were decreased to ~10<SUP>6</SUP> and 10<SUP>3</SUP>pmoA gene copy number g-DCW<SUP>-1</SUP> after MT and H<SUB>2</SUB>S degradation, respectively; however, both the amount of the DNA and mRNA transcript recovered their initial levels of ~10<SUP>7</SUP> and 10<SUP>5</SUP>pmoA gene copy number g-DCW<SUP>-1</SUP> after methane oxidation, respectively. The gene expression results indicate that the pmoA gene could be rapidly reproducible after methane oxidation. This study provides comprehensive information of kinetic interactions between methane and sulfur compounds.

New emergence pattern with variant porcine epidemic diarrhea viruses, South Korea, 2012-2015

Chung, H.C.,Lee, J.H.,Nguyen, V.G.,Huynh, T.M.L.,Lee, G.E.,Moon, H.J.,Park, S.J.,Kim, H.K.,Park, B.K. ELSEVIER 2016 VIRUS RESEARCH Vol.226 No.-

Since outbreaks of porcine epidemic diarrhea virus (PEDV) in the United States in 2013, explosive outbreaks of PED in South Korea have infected all age groups of pigs in 2014-2015year. This study analyzed a large collection of the Spike protein coding gene to infer the spatial-temporal diffusion history of PEDV. The studying results suggested that PEDVs in Korea belonged to different genogroups. While classical G1 was continuingly circulating between provinces of Korea, the pandemic G2a were recently introduced from China and USA. By the application of Bayesian phylogeographical analysis, this study demonstrated the spatial-temporal transmission of PEDVs within Korea. Of the recent emerged G2a viruses, J3142 strains showed potential recombination breakpoint (376-2,143nt) of S1 gene between KNU1303_Korea strain_G2a (KJ451046) and 45RWVCF0712_Thailand strain_G2b (KF724935). The pandemic G2a virus was partial neutralized by the antibodies invoked by the G1- based PED vaccine virus.

Linear accelertor를 이용한 뇌정위방사선수술(radiosurgery)

문재곤,정상섭,윤도흠,김선호,오성훈 한국의학사 1988 診斷과治療 Vol.8 No.12