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.

Effect of Genetic and Non-Genetic Factors Other Then Disease on Kid Survivability in Goat

Miah, S. G.,Husain, M. S.,Hoque, D. A.,Baik, D. H. 한국동물자원과학회 2002 한국축산학회지 Vol.44 No.3

이 연구는 벵갈 종과 그 교잡종 산양의 번식에 있어서 생시부터 90일령 까지의 생존율에 영향을 미치는 유전적 및 비 유전적인 요인을 규명하기 위하여 수행되었다. 분석에 이용된 모델은 지역, 교배조합, 자양의 성, 출생시 산자형태(1두, 2두, 3두 및 4두), 출생계절(여름 3월∼6월; 우기 7월∼10월; 및 겨울 11월∼2월), 출생연도와 2요인 상호작용의 효과를 포함하였다. 교잡종에 비하여 순종의 생존율은 31일령과 90일령 사이에 현저히 낮은 것으로 유의성(p<0.05)을 나타냈다. 생존율에 대한 지역의 효과는 16∼60일령 기간을 제외하고는 통계적으로 유의적인 차이를 보이지 않았다. 생존율에 대한 성의 효과는 초기 0∼7일령의 기간을 제외하고는 모두 통계적 유의성이 인정되었다. 생존율은 여름에 출생한 것이 가장 높았고 겨울과 우기(rainy season)의 순서로 나타났다. 16일령부터 60일령 사이의 자양의 생존율은 어미의 비유량에 의하여 영향을 받는 것으로 (p<0.05) 나타났고, 400∼600g/day 의 비유량인 경우에 생존율이 가장 높았던 반면 비유량이 80∼200g/day인 어미의 자양이 가장 낮은 생존율을 보였다. 생시체중은 90일령 까지의 모든 성장단계에서 자양의 생존율에 영향을 미치는 주요 요인이었다. 생존율은 모양의 비유량과 그리고 자양의 생시체중과 정의 상관관계를 보였다. 성과 생산지역간 (p<0.01) 또는 성과 출생시 산자형태간(p<0.05)의 상호작용은 통계적 유의성이 있었던 반면에 성과 유전적 그룹간의 상호작용은 유의성이 없었다. This experiment was conducted to investigate the genetic and non-genetic factors affecting kid survivability in goats from birth to 90 days of age. The purebreds had lower survivability than the crossbreds, with significant (p<0.05) difference amongst themselvess for the age of 31 to 90 days. The locational effect on survivability was insignificant for all the periods except 16 to 60 days (p<0.05) of age. The sex and birth type also had significant (p<0.05) effect on survivability for all the periods except 0 to 7 days of age. The survivability was found to be significantly (p<0.05) higher for kids born in summer season followed by those born in winter and rainy season. Milk produced by the does were significantly (p<0.05) affected on the survivability of kids during the period from 16 to 60 days of age. The survivability of kids were highest and lowest having milk yield of doe found to be 400∼600 g/day and 80∼200 g/day, respectively. Birth weight had significant (p<0.05) effect on survivability for all the stages of growth up to 90 days of age. Survivability was positively correlated with does' milk yield as well as kids birth weight. Interactions of sex with location or birth type were significant (p<0.01 and p<0.05, respectively) though interaction between sex and genetic group was insignificant.

Biochemical analysis of recombinant CYP4A11 allelic variant enzymes: W126R, K276T and S353G

Han, S.,Cha, G.S.,Chun, Y.J.,Lee, C.H.,Kim, D.,Yun, C.H. JAPANESE SOCIETY FOR THE STUDY OF XENOBIOTICS 2016 DRUG METABOLISM AND PHARMACOKINETICS Vol.31 No.6

Human CYP4A11 is the major ω-hydroxylase of fatty acids in the liver and kidneys. It produces 20-hydroxyeicosatetraenoic acid as well as hydroxylates fatty acids. In this study, we investigated the biochemical properties of three alleles of CYP4A11: W126R, K276T, and S353G. Site-directed mutagenesis of the wild type CYP4A11 was performed, to construct the W126R, K276T, and S353G variant clones. The CYP4A11 wild type and variant constructs were heterologously expressed in Escherichia coli. CO-binding spectra showed the expression of the wild type, K276T and S353G variants, indicating the functional P450 holoenzyme. The W126R variant was not expressed in E. coli. Binding affinities of lauric acid in K276T and S353G variants were stronger than that of wild type. Steady-state kinetics in the hydroxylation reaction of fatty acids were studied. The catalytic efficiencies (k<SUB>cat</SUB>/K<SUB>m</SUB>) of K276T and S353G variants in the reactions without cytochrome b<SUB>5</SUB> were approximately 2- and 4-fold higher, respectively, than that of wild type, and in the reactions with cytochrome b<SUB>5</SUB> they were approximately 2- and 3-fold higher, respectively. These results suggest that individuals carrying the alleles, K276T and S353G, might exhibit higher catalysis of CYP4A11, which may affect the endogenous metabolic products associated with regulation of blood pressure.

The gep oncogenes, Gα₁₂ and Gα₁₃, upregulate the transforming growth factor-β1 gene

Lee, S J,Yang, J W,Cho, I J,Kim, W D,Cho, M K,Lee, C H,Kim, S G Macmillan Publishers Limited 2009 Oncogene Vol.28 No.9

Transforming growth factor-β1 (TGFβ1) plays a role in neoplastic transformation and transdifferentiation. Gα<SUB>12</SUB> and Gα<SUB>13</SUB>, referred to as the gep oncogenes, stimulate mitogenic pathways. Nonetheless, no information is available regarding their roles in the regulation of the TGFβ1 gene and the molecules linking them to gene transcription. Knockdown or knockout experiments using murine embryonic fibroblasts and hepatic stellate cells indicated that a Gα<SUB>12</SUB> and Gα<SUB>13</SUB> deficiency reduced constitutive, auto-stimulatory or thrombin-inducible TGFβ1 gene expression. In contrast, transfection of activated mutants of Gα<SUB>12</SUB> and Gα<SUB>13</SUB> enabled the knockout cells to promote TGFβ1 induction. A promoter deletion analysis suggested that activating protein 1 (AP-1) plays a role in TGFβ1 gene transactivation, which was corroborated by the observation that a deficiency of the G-proteins decreased the AP-1 activity, whereas their activation enhanced it. Moreover, mutation of the AP-1-binding site abrogated the ability of Gα<SUB>12</SUB> and Gα<SUB>13</SUB> to induce the TGFβ1 gene. Transfection of a dominant-negative mutant of Rho or Rac, but not Cdc42, prevented gene transactivation and decreased AP-1 activity downstream of Gα<SUB>12</SUB> and Gα<SUB>13</SUB>. In summary, Gα<SUB>12</SUB> and Gα<SUB>13</SUB> regulate the expression of the TGFβ1 gene through an increase in Rho/Rac-dependent AP-1 activity, implying that the G-protein-coupled receptor (GPCR)-Gα<SUB>12</SUB> pathway is involved in the TGFβ1-mediated transdifferentiation process.Oncogene (2009) 28, 1230–1240; doi:10.1038/onc.2008.488; published online 19 January 2009

콩 탈곡기를 활용한 들깨 탈곡 기계화 연구

김형곤 ( H. G. Kim ),전현종 ( H. J. Jun ),김진구 ( J. G. Kim ),유승화 ( S. H. Yu ),김영근 ( Y. K. Kim ),강태경 ( T. G. Kang ),최일수 ( I. S. Choi ),이상희 ( S. H. Lee ),최용 ( Y. Choi ) 한국농업기계학회 2017 한국농업기계학회 학술발표논문집 Vol.22 No.2

들깨 생산을 위한 농작업에서 경운· 정지, 피복, 방제 등은 기계화 되었으나 파종, 정식, 예취· 탈곡은 기계화가 미흡한 실정이다. 본 연구에서는 기계화가 미흡한 들깨의 탈곡작업 기계화 촉진 및 범용화를 통한 들깨 기계화기술을 향상 하고자 수행하였다. 첫 번째 수행내용은 기존 콩 탈곡기 송풍 팬에 의해 손실되는 들깨를 줄이고자 송풍 팬 속도(450~1350 RPM)에 따른 풍구속도(m/s) 및 손실량(g)을 측정하기 위해 탈곡된 들깨 500 g을 탈곡기에 투입하여 실험하였다. 두 번째 수행내용은 투입된 작물의 줄기가 탈곡 시 배출이 용이하도록 탈곡기 덮개 안쪽에 가이드를 설치하여 탈곡에 미치는 영향을 실험하였다. 탈곡시험은 가이드를 설치하지 않은 경우와 설치한 경우로 분리하여 비교시험을 하였다. 가이드는 급동 회전방향과 10°, 20°, 30°를 유지하면서 배출구까지 도달하도록 설치하였고, 각도에 따른 잔물배출시간(s), 잔물배출무게(g) 및 잔물 파쇄 길이(cm)을 측정하였다. 시험결과 들깨의 손실률이 발생되는 구간은 1050 RPM, 풍구속도 5.27 m/s였으며 송풍 팬 회전속도가 증가할수록 손실률은 급격하게 증가하였다. 이는 실험 탈곡기 구조의 들깨 종말속도가 대략 5m/s로 송풍팬 회전속도를 1000 RPM, 풍구속도를 5m/s 이하로 설정해야 할 것으로 판단된다. 가이드 각도에 따른 잔물배출시간은 각도가 클수록 잔물배출시간이 단축되었고 특히 10°(18.90 s)는 무 설치(29.23 s) 보다 약 10초 단축되어 가이드 설치가 잔물배출에 효과적으로 나타났다. 잔물배출무게는 가이드 각도에 영향을 받지 않은 반면 잔물 파쇄 길이는 가이드 각도가 클수록 길게 나타났다. 결과적으로 실험 콩 탈곡기 적용 들깨 탈곡 시 5 m/s 이하의 풍속에서 잔물배출시간과 파쇄정도에 따른 들깨 손실 등을 고려한 복합적인 연구가 필요할 것으로 판단되었다.

New Evidence of Alleles (V199I and G52S) at the PRKAG3 (RN) Locus Affecting Pork Meat Quality

Chen, J.F.,Dai, L.H.,Peng, J.,Li, J.L.,Zheng, R.,Zuo, B.,Li, F.E.,Liu, M.,Yue, K.,Lei, M.G.,Xiong, Y.Z.,Deng, C.Y.,Jiang, S.W. Asian Australasian Association of Animal Productio 2008 Animal Bioscience Vol.21 No.4

The porcine PRKAG3 (RN) gene encodes the regulatory gamma subunit of adenosine monophosphate-activated protein kinase (AMPK), which is a good candidate gene affecting meat quality. In this study, the effects of two missense mutations A595G (Ile199Val) and G154A (Gly52Ser) in porcine PRKAG3 gene on meat quality traits were studied in M. Longissimus dorsi (LD), M. Semispinalis capitis (SC) and M. Biceps femoris (BF) from different populations of 326 pigs. The PRKAG3 alleles 199I, 199IV, 52S and 52G were identified with PCR-RFLPs and all genotypes - 199I/199I, 199I/199V, 199V/199V, 52S/52S, 52S/52G and 52G/52G - were found. The frequency of V allele was larger than that of I allele in all populations. I allele frequency was zero in Chinese Meishan pigs (population D) especially. G allele frequency was larger than that of S allele in all populations except Large White (population A). Both variations at the PRKAG3 locus significantly affected these meat quality traits. The pork meat quality has not previously been established in Meishan or crosses thereof. The results suggested that generally pH of LD, SC and BF was higher in Meishan pigs than that in other populations. Moreover, Meishan pigs showed higher water-holding capacity and intramuscular fat (IMF), lower water content and water loss percentage compared to other populations in terms of the two variations. The results present here supply new evidence that alleles V199I and G52S at the PRKAG3 locus affect pork meat quality and provide useful information on pork production.

HCV : PE-134 ; Hemoglobin decline during peginterferon Alfa-2B (PEG-2B)/ribavirin (RBV) treatment in real-life is associated with favorable SVR rates in difficult-to-treat patients with HCV genotype 1 (G1) infection

( G Teuber ),( S Mauss ),( D Huppe ),( E Zehnter ),( M P Manns ),( T Dahhan ),( U Meyer ),( T Witthoft ),( B Moller ),( N Dikopoulos ),( J Brack ),( B Stade ),( M Bilzer ),( Bng Hepatitis Study Group 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.1

Background and Aims: Recently, it has been shown for the overall G1 population that anemia as well as the maximal hemoglobin (Hb) decline during peginterferon/RBV treatment is associated with higher SVR rates. We here investigated whether the maximal Hb decline influences SVR rates in difficult-to-treat patients undergoing Peg2b/RBV therapy for HCV G1 infection in real-life. Methods: Data of patients treated for G1 infection within the German Peg2b/RBV observational study were retrospectively analyzed. In this real-life cohort study G1 infection was treated with Peg2b 1.5 μg/kg/wk + weight-based RBV (800-1200 mg/day) for up to 48 wks at 285 sites. Subjects who discontinued for non-response or for any other reasons were included in the analysis. SVR was defined as undetectable serum HCV-RNA 24 wks after EOT response. Only one patient received erythropoietin treatment for anemia. Results: 1851 patients had baseline and at least one Hb measurement during therapy. Overall SVR rate was 42.6% (789/1851). SVR rates were only slightly higher for subjects with an absolute Hb decline >3 g/dL (44.3%, 493/1114) compared to those with maximum Hb declines <3 g/dL (40.2%, 296/737) (p=0.08). In contrast, a significant (p=0.0004) difference in SVR rates was obtained by comparing subjects with Hb declines >2 g/dL (44.6%, 673/1510) with those who experienced Hb declines <2 g/dL (34.0%, 116/341). Similar SVR rates of 46.1% (164/356) and 44.1% (509/1154) in patients with Hb declines >2 g/dL even if they did/did not become anemic (Hb<10 g/dL) strongly support Hb decline, and not anemia, as primary beneficial mechanism improving SVR. As summarized in the table, Hb declines >2 g/dl were significantly associated with higher SVR rates in difficult-to-treat patients, such as subjects elder than 50 years or subjects with high baseline viral load >600.000 IU/ml. Interestingly no beneficial effect was observed in patients with low platelet count (<150/nL), an indicator of advanced fibrosis/cirrhosis. Patients who first developed a Hb decline >2 g/dL during weeks 0-4 were likely to achieve similar SVR (41.3%, 365/883) than those who developed a Hb decline <2 g/dL (44.9%, 386/859). In contrast, a Hb decline >2 g/dL compared to <2 g/dL during weeks 0-4 was associated with a 2-3 fold higher risk of anemia in female (16.6% vs 40.5%) and male patients (7.3% vs 19.0%) when compared with a Hb decline <2 g/dL. Conclusions: Patients with HCV genotype 1 infection and in particular the subgroup of difficult-to-treat patients elder than 50 years or with HVL, achieve up to 15% higher SVR rates when they develop a Hb decline >2 g/dL during Peg2b/RBV therapy. However, patients with low platelet count <150/nL do not achieve this beneficial virologic effect.

Involvement of S6K1 in mitochondria function and structure in HeLa cells

Park, J.,Tran, Q.,Mun, K.,Masuda, K.,Kwon, S.H.,Kim, S.H.,Kim, D.H.,Thomas, G.,Park, J. Pergamon Press ; Elsevier Science Ltd 2016 Cellular signalling Vol.28 No.12

The major biological function of mitochondria is to generate cellular energy through oxidative phosphorylation. Apart from cellular respiration, mitochondria also play a key role in signaling processes, including aging and cancer metabolism. It has been shown that S6K1-knockout mice are resistant to obesity due to enhanced beta-oxidation, with an increased number of large mitochondria. Therefore, in this report, the possible involvement of S6K1 in regulating mitochondria dynamics and function has been investigated in stable lenti-shS6K1-HeLa cells. Interestingly, S6K1-stably depleted HeLa cells showed phenotypical changes in mitochondria morphology. This observation was further confirmed by detailed image analysis of mitochondria shape. Corresponding molecular changes were also observed in these cells, such as the induction of mitochondrial fission proteins (Drp1 and Fis1). Oxygen consumption is elevated in S6K1-depeleted HeLa cells and FL5.12 cells. In addition, S6K1 depletion leads to enhancement of ATP production in cytoplasm and mitochondria. However, the relative ratio of mitochondrial ATP to cytoplasmic ATP is actually decreased in lenti-shS6K1-HeLa cells compared to control cells. Lastly, induction of mitophagy was found in lenti-shS6K1-HeLa cells with corresponding changes of mitochondria shape on electron microscope analysis. Taken together, our results indicate that S6K1 is involved in the regulation of mitochondria morphology and function in HeLa cells. This study will provide novel insights into S6K1 function in mitochondria-mediated cellular signaling.

S6K1 Phosphorylation of H2B Mediates EZH2 Trimethylation of H3: A Determinant of Early Adipogenesis

Yi, S.,Um, S.,Lee, J.,Yoo, J.,Bang, S.,Park, E.,Lee, M.,Nam, K.,Jeon, Y.,Park, J.,You, J.,Lee, S.J.,Bae, G.U.,Rhie, J.,Kozma, Sara C.,Thomas, G.,Han, J.W. Cell Press 2016 Molecular Cell Vol.62 No.3

S6K1 has been implicated in a number of key metabolic responses, which contribute to obesity. Critical among these is the control of a transcriptional program required for the commitment of mesenchymal stem cells to the adipocytic lineage. However, in contrast to its role in the cytosol, the functions and targets of nuclear S6K1 are unknown. Here, we show that adipogenic stimuli trigger nuclear translocation of S6K1, leading to H2BS36 phosphorylation and recruitment of EZH2 to H3, which mediates H3K27 trimethylation. This blocks Wnt gene expression, inducing the upregulation of PPARγ and Cebpa and driving increased adipogenesis. Consistent with this finding, white adipose tissue from S6K1-deficient mice exhibits no detectable H2BS36 phosphorylation or H3K27 trimethylation, whereas both responses are highly elevated in obese humans or in mice fed a high-fat diet. These findings define an S6K1-dependent mechanism in early adipogenesis, contributing to the promotion of obesity.

HCV, Alcoholic : PE-134 ; Hemoglobin decline during peginterferon Alfa-2B (PEG-2B)/ribavirin (RBV) treatment in real-Life is associated with favorable SVR rates in difficult-to-treat patients with HCV genotype 1 (G1) infection

( G Teuber ),( S Mauss ),( D Huppe ),( E Zehnter ),( M P Manns ),( T Dahhan6 ),( U Meyer ),( T Witthoft ),( B Moller9,),( N Dikopoulos ),( J Brack ),( B Stade ),( M Bilzer ),( The Bng Hepatitis Study 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.-

Background and Aims: Recently, it has been shown for the overall G1 population that anemia as well as the maximal hemoglobin (Hb) decline during peginterferon/RBV treatment is associated with higher SVR rates. We here investigated whether the maximal Hb decline influences SVR rates in difficult-to-treat patients undergoing Peg2b/RBV therapy for HCV G1 infection in real-life. Methods: Data of patients treated for G1 infection within the German Peg2b/RBV observational study were retrospectively analyzed. In this real-life cohort study G1 infection was treated with Peg2b 1.5 μg/kg/wk + weight-based RBV (800-1200 mg/day) for up to 48 wks at 285 sites. Subjects who discontinued for non-response or for any other reasons were included in the analysis. SVR was defined as undetectable serum HCV-RNA 24 wks after EOT response. Only one patient received erythropoietin treatment for anemia. Results: 1851 patients had baseline and at least one Hb measurement during therapy. Overall SVR rate was 42.6% (789/1851). SVR rates were only slightly higher for subjects with an absolute Hb decline >3 g/dL (44.3%, 493/1114) compared to those with maximum Hb declines <3 g/dL (40.2%, 296/737) (p=0.08). In contrast, a significant (p=0.0004) difference in SVR rates was obtained by comparing subjects with Hb declines >2 g/dL (44.6%, 673/1510) with those who experienced Hb declines <2 g/dL (34.0%, 116/341). Similar SVR rates of 46.1% (164/356) and 44.1% (509/1154) in patients with Hb declines >2 g/dL even if they did/did not become anemic (Hb<10 g/dL) strongly support Hb decline, and not anemia, as primary beneficial mechanism improving SVR. As summarized in the table, Hb declines >2 g/dl were significantly associated with higher SVR rates in difficult-to-treat patients, such as subjects elder than 50 years or subjects with high baseline viral load >600.000 IU/ml. Interestingly no beneficial effect was observed in patients with low platelet count (<150/nL), an indicator of advanced fibrosis/cirrhosis. Patients who first developed a Hb decline >2 g/dL during weeks 0-4 were likely to achieve similar SVR (41.3%, 365/883) than those who developed a Hb decline <2 g/dL (44.9%, 386/859). In contrast, a Hb decline >2 g/dL compared to <2 g/dL during weeks 0-4 was associated with a 2-3 fold higher risk of anemia in female (16.6% vs 40.5%) and male patients (7.3% vs 19.0%) when compared with a Hb decline <2 g/dL. Conclusions: Patients with HCV genotype 1 infection and in particular the subgroup of difficult-to-treat patients elder than 50 years or with HVL, achieve up to 15% higher SVR rates when they develop a Hb decline >2 g/dL during Peg2b/RBV therapy. However, patients with low platelet count <150/nL do not achieve this beneficial virologic effect.