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WWC1 and NF2 Prevent the Development of Intrahepatic Cholangiocarcinoma by Regulating YAP/TAZ Activity through LATS in Mice

  • Park, Jaeoh;Kim, Jeong Sik;Nahm, Ji Hae;Kim, Sang-Kyum;Lee, Da-Hye;Lim, Dae-Sik
    • Molecules and Cells
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    • v.43 no.5
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    • pp.491-499
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    • 2020
  • Hippo signaling acts as a tumor suppressor pathway by inhibiting the proliferation of adult stem cells and progenitor cells in various organs. Liver-specific deletion of Hippo pathway components in mice induces liver cancer development through activation of the transcriptional coactivators, YAP and TAZ, which exhibit nuclear enrichment and are activated in numerous types of cancer. The upstream-most regulators of Warts, the Drosophila ortholog of mammalian LATS1/2, are Kibra, Expanded, and Merlin. However, the roles of the corresponding mammalian orthologs, WWC1, FRMD6 and NF2, in the regulation of LATS1/2 activity and liver tumorigenesis in vivo are not fully understood. Here, we show that deletion of both Wwc1 and Nf2 in the liver accelerates intrahepatic cholangiocarcinoma (iCCA) development through activation of YAP/TAZ. Additionally, biliary epithelial cell-specific deletion of both Lats1 and Lats2 using a Sox9-CreERT2 system resulted in iCCA development through hyperactivation of YAP/TAZ. These findings suggest that WWC1 and NF2 cooperate to promote suppression of cholangiocarcinoma development by inhibiting the oncogenic activity of YAP/TAZ via LATS1/2.

Developmental Characteristics of Cloned Embryos Reconstructed with Induced Pluripotent Stem Cells in Pigs (돼지 유도만능줄기세포 유래 복제란의 특성 분석)

  • Kwon, Dae-Jin;Oh, Jae-Don;Park, Mi-Ryung;Hwang, In-Sul;Park, Eung Woo;Hwang, Seongsoo
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.3
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    • pp.232-239
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    • 2019
  • In general, cloned pigs have been produced using the somatic cell nuclear transfer (SCNT) technique with various types of somatic cells; however, the SCNT technique has disadvantages not only in its low efficiency but also in the development of abnormal clones. This study aimed to compare early embryonic development and quality of SCNT embryos with those of induced pluripotent stem cells (iPSCs) NT embryos (iPSC-NTs). Ear fibroblast cells were used as donor cells and iPSCs were generated from these cells by lentiviral transduction with human six factors (Oct4, Sox2, c-Myc, Nanog, Klf4 and Lin28). Blastocyst formation rate in iPSC-NT (23/258, 8.9%) was significantly lower than that in SCNT (46/175, 26.3%; p < 0.05). Total cell number in blastocysts was similar between two groups, but blastocysts in iPSC-NT had a lower number of apoptotic cells than in SCNT (2.0 ± 0.6 vs. 9.8 ± 2.9, p < 0.05). Quantitative PCR data showed that apoptosis-related genes (bax, caspase-3, and caspase-9) were highly expressed in SCNT than iPSC-NT (p < 0.05). Although an early development rate was low in iPSC-NT, the quality of cloned embryos from porcine iPSC was higher than that of embryos from somatic cells. Therefore, porcine iPSCs could be used as a preferable cell source to create a clone or transgenic animals by using the NT technique.

The Effect to Agricultural Crop Yield and Sulfur Content in Leaves by Sulfur Dioxide Gas Emission from Onsan Nonferric Metal Industrial Complex (온산(溫山) 공단지역(工團地域)의 배출(排出)가스 (아황산(亞黃酸))가 농작물(農作物) 수량(收量) 및 식물체내(植物體內) 유황함량(硫黃含量)에 미치는 영향(影響))

  • Shin, Nam-Chul
    • Korean Journal of Environmental Agriculture
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    • v.4 no.1
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    • pp.52-56
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    • 1985
  • This study was undertaken to investigate the crop yield of pear and rice cultivated around Onsan Industrial complex, the sulfur content in their leaves, and the crop loss. The correlation between them is as follows: 1) The sulfur content in pear and rice leaves in studied area is higher than that in unpolluted area, and there is crop loss in studied area. 2) There is a good correlation between sulfur content and crop loss of pear and rice. The air pollution in studied area is considered to be the reason of the crop loss. 3) The regression equation between the sulfur content in crop leaves and the crop loss seems to be useful for the crop loss rate by sulfur dioxide in the damaged area and that seems to be a good reference for the compensation of pear and rice loss by sulfur dioxide gas emission from Onsan Industral Complex.

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Expression of C6orf62 in Human Embryonic Stem Cells and Cancer Cells (인간 배아 줄기세포와 암 세포에서의 C6orf62의 발현 패턴)

  • Yoo, Han-Na;Yoo, Jung-Ki;Choi, Seoung-Jun;Kim, Jin-Kyeoung
    • Reproductive and Developmental Biology
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    • v.34 no.3
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    • pp.229-233
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    • 2010
  • Pluripotency and self-renewal capacity of human embryonic stem cells (hESCs) are retained by hESCs related genes as OCT4, SOX2 and NANOG. These genes are shown high expression level in diverse cancer cells and have potential role in the carcinogenesis. On the contrary to this, several genes which are up-regulated in the differentiated hESCs are involved to suppress the carcinogenesis or proliferation of cells. We discovered several genes in immortalized lung fibroblast (WI-38 VA13) by suppression subtractive hybridization. Among them, we focused chromosome 6 open reading frame 62 (C6orf62) which is uncharacterized, mapped to 6p22.3 and generated to Hepatitis B virus X-transactivated proteins (HBVx-transactivated proteins, XTP). Aim of this study was to characterize C6orf62 through analyzing of expression pattern in various cell lines. Expression of C6orf62 was significantly upregulated in diverse normal cell lines than cancer cell lines. And C6orf62 was up-regulated in differentiated hESCs (endothelial cells, neural cells) compared to those of undifferentiated hESCs. Also, C6orf62 in WI-38 cells was highly up-regulated during G1/S transition of the cell cycle. Taken together, C6orf62 is shown expression pattern similar to differentiated hESCs-associated genes which down-regulated in cancer cells. Therefore, we assume that C6orf62 may participate to suppress the proliferation and to induce differentiation through regulating the cell cycle.

Pyrolysis oil refining by Fly-ash absorption (Fly-ash 흡착기법을 이용한 열분해유 정제)

  • Im, EunJung;Kim, SungHyun;Chun, ByungHee;SunWoo, Hwan;Jeong, IckCheol
    • 한국신재생에너지학회:학술대회논문집
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    • 2011.05a
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    • pp.222-222
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    • 2011
  • Plastic product is increasing by the growth of its demand and most of refused plastics are incinerated or reclaimed. However, the refused plastic is not easily decomposed and has the environmental problem with its various toxic gas in case of incineration. Therefore, many countries such as USA, Japan, Germany and other developed industrial countries as well as Korea are interested in studying the recyclable resource of refused plastic. The macromolecular waste pyrolysis has the advantage of collecting of raw materials in high price and can at least get fuel gas or oil with high heat capacity. It also discharges low waste gas and low toxic gas including SOx, NOx and HCl heavy metals. However, pyrolyzed oil includes enough excess unsaturated hydrocarbons to form tar, which can cause the nozzle of engines to plug when pyrolyzed oil is used as fuel. Activated carbon was proven to have prominent adsorption capability among the other adsorbents that were mainly composed of carbon. This study examined the possibility of application in activated charcoal of its solid formation by analysing the feature of pyrolysis which is one of the chemical recycling methods and getting chemical analysis of the product and activated energy. Analyze the element of the oil produced by pyrolysis using GC-MS. The experiment of tar adsorption using fly-ash showed that fly-ash improved the optical intensity of pyrolyzed oil and decreased oxygen compounds in the pyrolyzed oil.

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Embryo Aggregation Promotes Derivation Efficiency of Outgrowths from Porcine Blastocysts

  • Lee, Sang-Goo;Park, Jin-Kyu;Choi, Kwang-Hwan;Son, Hye-Young;Lee, Chang-Kyu
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.11
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    • pp.1565-1572
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    • 2015
  • Porcine embryonic stem cells (pESCs) have become an advantageous experimental tool for developing therapeutic applications and producing transgenic animals. However, despite numerous reports of putative pESC lines, deriving validated pESC lines from embryos produced in vitro remains difficult. Here, we report that embryo aggregation was useful for deriving pESCs from in vitro-produced embryos. Blastocysts derived from embryo aggregation formed a larger number of colonies and maintained cell culture stability. Our derived cell lines demonstrated expression of pluripotent markers (alkaline phosphatase, Oct4, Sox2, and Nanog), an ability to form embryoid bodies, and the capacity to differentiate into the three germ layers. A cytogenetic analysis of these cells revealed that all lines derived from aggregated blastocysts had normal female and male karyotypes. These results demonstrate that embryo aggregation could be a useful technique to improve the efficiency of deriving ESCs from in vitro-fertilized pig embryos, studying early development, and deriving pluripotent ESCs in vitro in other mammals.

Porcine somatic cell nuclear transfer using telomerase reverse transcriptase-transfected mesenchymal stem cells reduces apoptosis induced by replicative senescence

  • Jeon, Ryounghoon;Rho, Gyu-Jin
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.3
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    • pp.215-222
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    • 2020
  • Mesenchymal stem cells (MSCs) have been widely used as donor cells for somatic cell nuclear transfer (SCNT) to increase the efficiency of embryo cloning. Since replicative senescence reduces the efficiency of embryo cloning in MSCs during in vitro expansion, transfection of telomerase reverse transcriptase (TERT) into MSCs has been used to suppress the replicative senescence. Here, TERT-transfected MSCs in comparison with early passage MSCs (eMSCs) and sham-transfected MSCs (sMSCs) were used to evaluate the effects of embryo cloning with SCNT in a porcine model. Cloned embryos from tMSC, eMSC, and sMSC groups were indistinguishable in their fusion rate, cleavage rate, total cell number, and gene expression levels of OCT4, SOX2 and NANOG during the blastocyst stage. The blastocyst formation rates of tMSC and sMSC groups were comparable but significantly lower than that of the eMSC group (p < 0.05). In contrast, tMSC and eMSC groups demonstrated significantly reduced apoptotic incidence (p < 0.05), and decreased BAX but increased BCL2 expression in the blastocyst stage compared to the sMSC group (p < 0.05). Therefore, MSCs transfected with telomerase reverse transcriptase do not affect the overall development of the cloned embryos in porcine SCNT, but enables to maintain embryo quality, similar to apoptotic events in SCNT embryos typically achieved by an early passage MSC. This finding offers a bioengineering strategy in improving the porcine cloned embryo quality.

Identification of Candidate Porcine miRNA-302/367 Cluster and Its Function in Somatic Cell Reprogramming

  • Son, Dong-Chan;Hwang, Jae Yeon;Lee, Chang-Kyu
    • Reproductive and Developmental Biology
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    • v.38 no.2
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    • pp.79-84
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    • 2014
  • MicroRNAs (miRNAs) are approximately 22 nucleotides of small noncoding RNAs that control gene expression at the posttranscriptional level through translational inhibition and destabilization of their target mRNAs. The miRNAs are phylogenetically conserved and have been shown to be instrumental in a wide variety of key biological processes including cell cycle regulation, apoptosis, metabolism, imprinting, and differentiation. Recently, a paper has shown that expression of the miRNA-302/367 cluster expressed abundantly in mouse and human embryonic stem cells (ESCs) can directly reprogram mouse and human somatic cells to induced pluripotent stem cells (iPSCs) efficiently in the absence of any of the four factors, Oct4, Sox2, c-Myc, and Klf4. To apply this efficient method to porcine, we analyzed porcine genomic sequence containing predicted porcine miRNA-302/367 cluster through ENSEMBL database, generated a non-replicative episomal vector system including miRNA-302/367 cluster originated from porcine embryonic fibroblasts (PEF), and tried to make porcine iPSCs by transfection of the miRNA-302/367 cluster. Colonies expressing EGFP and forming compact shape were found, but they were not established as iPSC lines. Our data in this study show that pig miRNA-302/367 cluster could not satisfy requirement of PEF reprogramming conditions for pluripotency. To make pig iPSC lines by miRNA, further studies on the role of miRNAs in pluripotency and new trials of transfection with conventional reprogramming factors are needed.

Identification of genes related to intramuscular fat content of pigs using genome-wide association study

  • Won, Sohyoung;Jung, Jaehoon;Park, Eungwoo;Kim, Heebal
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.2
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    • pp.157-162
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    • 2018
  • Objective: The aim of this study is to identify single nucleotide polymorphisms (SNPs) and genes related to pig IMF and estimate the heritability of intramuscular fat content (IMF). Methods: Genome-wide association study (GWAS) on 704 inbred Berkshires was performed for IMF. To consider the inbreeding among samples, associations of the SNPs with IMF were tested as random effects in a mixed linear model using the genetic relationship matrix by GEMMA. Significant genes were compared with reported pig IMF quantitative trait loci (QTL) regions and functional classification of the identified genes were also performed. Heritability of IMF was estimated by GCTA tool. Results: Total 365 SNPs were found to be significant from a cutoff of p-value <0.01 and the 365 significant SNPs were annotated across 120 genes. Twenty five genes were on pig IMF QTL regions. Bone morphogenetic protein-binding endothelial cell precursor-derived regulator, forkhead box protein O1, ectodysplasin A receptor, ring finger protein 149, cluster of differentiation, tyrosine-protein phosphatase non-receptor type 1, SRY (sex determining region Y)-box 9 (SOX9), MYC proto-oncogene, and macrophage migration inhibitory factor were related to mitogen-activated protein kinase pathway, which regulates the differentiation to adipocytes. These genes and the genes mapped on QTLs could be the candidate genes affecting IMF. Heritability of IMF was estimated as 0.52, which was relatively high, suggesting that a considerable portion of the total variance of IMF is explained by the SNP information. Conclusion: Our results can contribute to breeding pigs with better IMF and therefore, producing pork with better sensory qualities.

Loss of Heterozygosity at the Calcium Regulation Gene Locus on Chromosome 10q in Human Pancreatic Cancer

  • Long, Jin;Zhang, Zhong-Bo;Liu, Zhe;Xu, Yuan-Hong;Ge, Chun-Lin
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2489-2493
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    • 2015
  • Background: Loss of heterozygosity (LOH) on chromosomal regions is crucial in tumor progression and this study aimed to identify genome-wide LOH in pancreatic cancer. Materials and Methods: Single-nucleotide polymorphism (SNP) profiling data GSE32682 of human pancreatic samples snap-frozen during surgery were downloaded from Gene Expression Omnibus database. Genotype console software was used to perform data processing. Candidate genes with LOH were screened based on the genotype calls, SNP loci of LOH and dbSNP database. Gene annotation was performed to identify the functions of candidate genes using NCBI (the National Center for Biotechnology Information) database, followed by Gene Ontology, INTERPRO, PFAM and SMART annotation and UCSC Genome Browser track to the unannotated genes using DAVID (the Database for Annotation, Visualization and Integration Discovery). Results: The candidate genes with LOH identified in this study were MCU, MICU1 and OIT3 on chromosome 10. MCU was found to encode a calcium transporter and MICU1 could encode an essential regulator of mitochondrial $Ca^{2+}$ uptake. OIT3 possibly correlated with calcium binding revealed by the annotation analyses and was regulated by a large number of transcription factors including STAT, SOX9, CREB, NF-kB, PPARG and p53. Conclusions: Global genomic analysis of SNPs identified MICU1, MCU and OIT3 with LOH on chromosome 10, implying involvement of these genes in progression of pancreatic cancer.