• Asian Pacific Journal of Cancer Prevention
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• 제17권3호
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• pp.1023-1035
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• 2016

The purpose of this study was to investigate the role of colon cancer stem cells (CSCs) during chemically-induced rat multi-step colon carcinogenesis with or without the treatment with a specific cyclooxygenase-2 inhibitor drug (celecoxib). Two experiments were performed, the first, a short term 12 week colon carcinogenesis bioassay in which only surrogate markers for colon cancer, aberrant crypt foci (ACF) lesions, were formed. The other experiment was a medium term colon cancer rat assay in which tumors had developed after 32 weeks. Treatment with celecoxib lowered the numbers of ACF, as well as the tumor volumes and multiplicities after 32 weeks. Immunohistochemical proliferating cell nuclear antigen (PCNA) labeling indexes LI (%) were downregulated after treatment by celecoxib. Also different cell surface antigens known to associate with CSCs such as the epithelial cell adhesion molecule (EpCAM), CD44 and CD133 were compared between the two experiments and showed differential expression patterns depending on the stage of carcinogenesis and treatment with celecoxib. Flow cytometric analysis demonstrated that the numbers of CD133 cells were increased in the colonic epithelium after 12 weeks while those of CD44 but not CD133 cells were increased after 32 weeks. Moreover, aldehyde dehydrogenase-1 activity levels in the colonic epithelium (a known CSC marker) detected by ELISA assay were found down-regulated after 12 weeks, but were up-regulated after 32 weeks. The data have also shown that the protective effect of celecoxib on these specific markers and populations of CSCs and on other molecular processes such as apoptosis targeted by this drug may vary depending on the genetic and phenotypic stages of carcinogenesis. Therefore, uncovering these distinction roles of CSCs during different phases of carcinogenesis and during specific treatment could be useful for targeted therapy.

• Proceedings of the Korean Society of Crop Science Conference
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.131-131
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• 2017

Though the Platycodon grandiflorum, has a broad range of pharmacologic properties, but the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two-dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (${\geq}2-fold$) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose-1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, the frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). Taken together, the protein profile may provide insight clues for better understanding the characteristics of proteins and its metabolic activities in various explants of this essential medicinal plant P. grandiflorum.

• Proceedings of the Korean Society of Crop Science Conference
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.125-125
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• 2017

The roots of Platycodon grandiflorum are commonly used for treating bronchitis, asthma, tuberculosis, diabetes, and other inflammatory diseases. Since the molecular mechanism underlying the roots of the plant is unclear. Therefore, the present study was conducted to profile proteins from liquid cultured tetraploid roots of Platycodon grandi orum fl using high throughput proteome approach. Two-dimensional gels stained with CBB, a total of 659 differentially expressed proteins were identified from the liquid medium cultured tetraploid roots of which 32 proteins spots (${\geq}1.5-fold$) were sorted for mass spectrometry analysis. Out of these 32 proteins, a total of 15 proteins were up-regulated such as Serine carboxypeptidase-like 27, Transcription factor bHLH150, 60 kDa jasmonate-induced protein, Cytosolic Fe-S cluster assembly factor NBP35, Regulatory associated protein of TOR 2 and a total of 17 proteins were down-regulated such as Protein G1-like2, Phenylalanine ammonia-lyase, Fructokinase-2, Trihelix transcription factor GT-3a, Guanine nucleotide-binding protein alpha-1 subunit. However, the frequency distribution of identified proteins was carried out within functional categories based on molecular functions, cellular components, and biological processes. Functional categorization revealed that the most of the identified proteins from the explants were mainly associated with the nucleic acid binding, oxidoreductase, transferase activity, protein binding and hydrolase activity. In addition, the proteomic feedback of tetraploid roots of P. grandiflorum may potentially be used to understand the characteristics of proteins and their functions.

• Journal of Plant Biotechnology
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• 제40권1호
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• pp.9-17
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• 2013

Ran is a small GTP-binding protein that binds and subsequently hydrolyzes GTP. The functions of Ran in nuclear transport and mitotic progression are well conserved in plants and animals. In animal cells, stress treatments cause Ran relocalization and slowing of nuclear transport, but the role of Ran proteins in plant cells exposed to stress is still unclear. We have therefore compared Ran genes from three EST libraries construed from different cell types of sweetpotato and the distribution pattern of Ran ESTs differed according to cell type. We further characterized two IbRan genes. IbRan1 is a specific EST to the suspension cells and leaf libraries, and IbRan2 is specific EST to the root library. IbRan1 showed 94.6 % identity with IbRan2 at the amino acid level, but the C-terminal region of IbRan1 differed from that of IbRan2. These two genes showed tissue-specific differential regulation in wounded tissues. Chilling stress induced a similar expression pattern in both IbRan genes in the leaves and petioles, but they were differently regulated in the roots. Hydrogen peroxide treatment highly stimulated IbRan2 mRNA expression in the leaves and petioles, but had no significant effect on IbRan1 gene expression. These results showed that the transcription of these two IbRan genes responds differentially to abiotic stresses and that they are subjected to tissue-specific regulation. Plant Ran-type small G-proteins are a multigenic family, and the characterization of each Ran genes under various environmental stresses will contribute toward our understanding of the distinctive function of each plant Ran isoform.

• Asian Pacific Journal of Cancer Prevention
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• 제14권10호
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• pp.5663-5669
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• 2013

The histone methyltransferase EZH2 (enhancer of zeste homolog 2) plays critical roles in prostate cancer (PCa) development and is a potential target for PCa treatment. Triptolide possesses anti-tumor activity, but it is unknown whether its therapeutic effect relates with EZH2 in PCa. Here we described EZH2 as a target for Triptolide in PCa cells. Our data showed that Triptolide suppressed PCa cell growth and reduced the expression of EZH2. Overexpression of EZH2 attenuated the Triptolide induced cell growth inhibition. Moreover, Triptolide treatment of PC-3 cells resulted in elevated mRNA levels of target genes (ADRB2, CDH1, CDKN2A and DAB2IP) negatively regulated by EZH2 as well as reduced mRNA levelsan of EZH2 positively regulated gene (cyclin D1). Our findings suggest the PCa cell growth inhibition mediated by Triptolide might be associated with downregulation of EZH2 expression and the subsequent modulation of target genes.

• Asian-Australasian Journal of Animal Sciences
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• 제25권6호
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• pp.789-793
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• 2012

Fertilization of the oocyte commences embryogenesis during which maternally inherited mRNAs are degraded and the embryonic genome is activated. Transcription of embryonic mRNA is initiated by embryonic genome activation (EGA). RNA polymerase II (RNA Pol II) is responsible for the synthesis of mRNAs and most small nuclear RNAs, and consists of 12 subunits, the largest of which characteristically harbors a unique C-terminal domain (CTD). Transcriptional activity of RNA Pol II is highly regulated, in particular, by phosphorylation of serine residues in the CTD. Here, we have shown the presence of RNA Pol II CTD phosphoisoforms in porcine oocytes and preimplantation embryos. The distribution pattern as well as phosphorylation dynamics in germinal vesicles and during embryogenesis differed in developmental stages with these isoforms, indicating a role of RNA Pol II CTD phosphorylation at the serine residue in transcriptional activation during both oocyte growth and embryonic genome activation. We additionally examined the effects of the RNA Pol II inhibitor, ${\alpha}$-amanitin, on embryo development. Our results show that inhibition of polymerase, even at very early stages and for a short period of time, dramatically impaired blastocyst formation. These findings collectively suggest that the functionality of maternal RNA Pol II, and consequently, expression of early genes regulated by this enzyme are essential for proper embryo development.

• Proceedings of the Microbiological Society of Korea Conference
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• 한국미생물학회 2007년도 International Meeting of the Microbiological Society of Korea
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• pp.83-85
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• 2007

Deep-sea bacteria are adapted to extreme environments, such as high pressures and cold temperatures. We have isolated many piezophiles which grow well even under high pressures from deep-sea sediment. Shewanella violacea DSS12 and Moritella japonica DSK1 have the ability to grow at up to 70 MPa, and those bacteria have unique mechanisms of gene expression in response to high pressure conditions. The combination of gene expression systems in piezophiles, like the high pressure-dependent promoters and GFP reporter gene, may reveal highly fluorescent cells when exposed to high hydrostatic pressure conditions. It is predicted that a novel bio-sensing system can be made to probe high pressure environments using living bacteria. First, gene transformation into our piezophiles, strains DSS12 and DSK1, were examined. Eschericha coli S17-1 was used for bacterial conjugation with those piezophiles. As a result, the broad host range vector, pKT231, and the shuttle vector, pTH10, were successfully introduced to DSS12 and DSK1, respectively. Next, The pressure regulated promoters from DSS12 and DSK1 were cloned into proper vectors and combined with GFP as a reporter gene downstream of each promoter. The transformants of DSK1 and DSS12 with the recombinant pTH10 and pKT231 plasmid, which has cadA and glnA promoters (each of them is a pressure regulated promoter from DSK1 and DSS12, respectively) and GFP, were grown under high pressure and gene expression of GFP promoted by 50 MPa pressure was confirmed. This is a critical point to create a pressure-sensing bacteria, as the "High Pressure Glow Cells", which will indicate the level of environmental pressure using fluorescence of GFP as a reporter gene.

• Journal of Microbiology and Biotechnology
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• 제18권1호
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• pp.80-87
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• 2008

The 3T3-L1 cell line is a well-established and commonly used in vitro model to assess adipocyte differentiation. Over the course of several days, confluent 3T3-L1 cells can be converted to adipocytes in the presence of an adipogenic cocktail. In this study, the effects of chitosan oligosaccharides (CO) on adipocyte differentiation of 3T3-L1 cells were studied. The CO significantly decreased lipid accumulation, a marker of adipogenesis, in a dose-dependent manner. The low molecular mass CO (1-3 kDa) were the most effective at inhibiting adipocyte differentiation. Moreover, mRNA expression levels of both CCAAT/enhancer-binding protein (C/EBP) ${\alpha}$ and peroxisome proliferator-activated receptor (PPAR) ${\gamma}$, the key adipogenic transcription factors, were markedly decreased by CO treatments. CO also significantly down regulated adipogenic marker proteins such as leptin, adiponectin, and resistin. Our results suggest a role for CO as antiobesity agents by inhibiting adipocyte differentiation mediated through the down regulated expression of adipogenic transcription factors and other specific genes.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 한국환경독성학회 2002년도 추계국제학술대회
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• pp.156-156
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• 2002

The goal of this study is to develop a biomarker used in monitoring abnormal behaviors of Japanese medaka (Oryzias latipes) as a model organism caused by hazardous chemicals that are toxic and persistent in the ecosystem. A widely used insecticide, diazinon (O, O-diethyl O- (2-isopropyl-4-methyl-6-pyrimidinyl) phosphorothioate), is highly neurotoxic to fish, and it is also well known that it causes vertebral malformation and behavioral changes of fish at relatively low concentrations. The fish behaviors were observed on a real time basis using an image processing and automatic data acquisition system. The genes potentially involved in the abnormal behaviors were cloned using suppression subtractive hybridization (SSH) technique. The untreated individuals showed common behavioral characteristics. When the test fish was affected by diazinon at a concentration of 0.1 and 1 ppm, some specific patterns were observed in its behavioral activity and locomotive tracks. The typical patterns were enhanced surfacing activity, opercular movement, erratic movement, tremors and convulsions as reported previously. The number of genes up-regulated tty diazinon treatment were 97 which includes 27 of unknown genes. The number of down-regulated genes were 99 including 60 of unknown genes. These gene expression patterns will be analyzed by the artificial neural networks such as self organization map (SOM) and multilayer perceptron (MLP), revealing the role of genes responsible for the behaviors. These results may provide molecular biological and neurobehavioral bases of a biomonitoring system for diazinon using a model organism such as fish.

• KOREAN JOURNAL OF CROP SCIENCE
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• 제60권1호
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• pp.97-106
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• 2015

Platycodon grandiflorum, commonly known as Doraji in Korea, has a wide range of pharmacologic properties, such as reducing adiposity and hyperlipidemia, and antiatherosclerotic effects. However, the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (${\geq}$ 2-fold) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose-1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). In that way, the exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.