• Title/Summary/Keyword: Clusterin

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Effects of Panax ginseng and Ziziphus jujuba on stress-induced apoptosis in rats

  • Kim, Hyung-Chan
    • International Journal of Oral Biology
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    • v.33 no.1
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    • pp.7-12
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    • 2008
  • PG has been well studied about effects of stress resistance. Although ZJ has been known that it had stress resistance effect since ancient times, its pharmacological properties and clinical applications have not been studied and reported until recently. Therefore, the purpose of this study is to determine whether effects of stress hormones, mechanism of stress protein could be induced by PG and ZJ of herb extract ingestion during stress exposure. In addition, this study identified expression of apoptosis factors related to stress. 1) Bcl-2 expression of the stressed rats decreased in comparison with the unstressed rats in heart and stomach. Bcl-2 expression of rats administered to PG was higher than the stressed rats in heart and that of rats administered to ZJ was higher than the stressed rats in stomach. 2) Stressed rats were decreased in p53 protein expression than normal rats. Thus, the results suggest stress-induced apoptosis is p53-independent apoptosis. And these results demonstrated that PG or ZJ administration helped to return from stress state to normal. 3) Clusterin expressed markedly in only salivary gland, but that of expression was no difference among four groups in tissues. Clusterin expression has no relation of stress-induced apoptosis.

Hypoxia Inducible Factor-1α Directly Regulates Nuclear Clusterin Transcription by Interacting with Hypoxia Response Elements in the Clusterin Promoter

  • Park, Jeongsook;Park, So Yun;Shin, Eunkyung;Lee, Sun Hee;Kim, Yoon Sook;Lee, Dong Hoon;Roh, Gu Seob;Kim, Hyun Joon;Kang, Sang Soo;Cho, Gyeong Jae;Jeong, Bo-Young;Kim, Hwajin;Choi, Wan Sung
    • Molecules and Cells
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    • v.37 no.2
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    • pp.178-186
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    • 2014
  • Differential transcription of the clusterin (CLU) gene yields two CLU isoforms, a nuclear form (nCLU) and a secretory form (sCLU), which play crucial roles in prostate tumorigenesis. Pro-apoptotic nCLU and anti-apoptotic sCLU have opposite effects and are differentially expressed in normal and cancer cells; however, their regulatory mechanisms at the transcriptional level are not yet known. Here, we examined the transcriptional regulation of nCLU in response to hypoxia. We identified three putative hypoxia response elements (HREs) in the human CLU promoter between positions -806 and +51 bp. Using a luciferase reporter, electrophoretic gel mobility shift, and chromatin immunoprecipitation assays, we further showed that hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$) bound directly to these sites and activated transcription. Exposure to the hypoxia-mimetic compound $CoCl_2$, incubation under 1% $O_2$ conditions, or overexpression of HIF-$1{\alpha}$ enhanced nCLU expression and induced apoptosis in human prostate cancer PC3M cells. However, LNCaP prostate cancer cells were resistant to hypoxia-induced cell death. Methylation-specific PCR analysis revealed that the CLU promoter in PC3M cells was not methylated; in contrast, the CLU promoter in LNCap cells was methylated. Co-treatment of LNCaP cells with $CoCl_2$ and a demethylating agent promoted apoptotic cell death through the induction of nCLU. We conclude that nCLU expression is regulated by direct binding of HIF-$1{\alpha}$ to HRE sites and is epigenetically controlled by methylation of its promoter region.

Identification of Differentially Expressed Genes by Proto-oncogene Protein DEK using Annealing Control Primers

  • Kim, Dong-Wook;Lee, Jae-Hwi;Seo, Sang-Beom
    • Biomolecules & Therapeutics
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    • v.16 no.3
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    • pp.184-189
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    • 2008
  • The proto-oncogene protein DEK has been implicated in various human disease including cancer. We have shown that DEK induces caspase-dependent apoptosis in Drosophila by regulating histone acetylation. Reverse transcription-polymerase chain reaction (RT-PCR) method based on annealing control primers was used to screen and identify differentially expressed genes (DEGs) in DEK overexpressed HeLa cells. Among the genes identified, clusterin and fibrillarin have major role in apoptosis pathway regulation. TFIIIC and RPS24 are implicated in HAT mediated transcriptional initiation and cololectal cancer, respectively. To further analyze DEK's role in apoptosis, multiplex PCR was performed. Caspase-3, -7, and -10 and proapoptotic gene bid were newly identified as possible target genes regulated by DEK expression.

A Study of Expanded Severity Index of Voltage Sag Using Fuzzy Clusterin (Fuzzy Clustering을 이용한 순간전압강하(Voltage Sag)의 확장된 심각도 지수(Expanded Severity Index) 연구)

  • Oh, Won-Wook;Kim, Yong-Su
    • Proceedings of the Korean Society of Computer Information Conference
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    • 2011.01a
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    • pp.81-84
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    • 2011
  • 본 논문은 전압 이벤트 현상 중 순간전압강하(Sag) 현상에 초점을 맞추었다. Sag 현상의 심각한 정도를 표현하는 심각도(Voltage Sag Severity) 지수는 동일 지속시간에 대한 임계치와의 비로 표현하였다. 제안하는 확장된 심각도(Expanded Severity) 지수는 sag현상의 분포에 따른 일시반복성의 정보를 표현하였다. 기존의 임계치를 표현하는 ITIC curve를 기반으로 된 심각도와 sag 현상이 발생하는 지속시간-전압 그래프의 분포를 fuzzy clustering을 통하여 medoid를 측정하고, medoid의 심각도와 실제 임계치에 근접한 sag 지점의 심각도를 계산하여 비교하였다. 확장된 심각도 지수는 심각도가 높은 현상들과의 연계성을 나타내는 지수로 심각한 정도의 수치 정보 이외에 일시적인 현상인지 지속 반복적인 현상인지를 0과 1사이의 수치로 표현하였고, 실험을 통하여 입증하였다.

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Proteomic analysis of Korean mothers' human milk at different lactation stages; postpartum 1, 3, and 6 weeks (출산 후 경과한 날에 따른 한국인 산모의 모유 단백체 분석)

  • Park, Jong-Moon;lee, Hookeun;Song, Seunghyun;Hahn, Won-Ho;Kim, Mijeong;Lee, Joohyun;Kang, Nam Mi
    • Analytical Science and Technology
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    • v.30 no.6
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    • pp.348-354
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    • 2017
  • In this study, patterns of proteome expression were monitored and specifically expressed proteins in human milk were detected in collected human milk after 1 week, 3 weeks, and 6 weeks from delivery. A quantitative shotgun proteomic approach was used to identify human milk proteins and reveal their relative expression amounts. For each sample, two independent human milk samples from two mothers were pooled, and then three replicated shotgun proteomic analyses were carried out. Casein, which is a highly abundant protein in human milk, was removed, and then trypsin was treated to produce a digested peptide mixture. The peptides were loaded in the home-made reversed-phase C18 fused-silica capillary column, and then the eluted peptides were analyzed by using a linear ion-trap mass spectrometer. The relative quantitation of proteins was performed by the normalized spectral count method. For each sample, 81-109 non-redundant proteins were identified. The identified proteins consisted of glycoproteins, metabolic enzyme, and chaperon enzymes such as lactoferrin, carboxylic ester hydrolase, and clusterin. The comparative analysis for the 63 proteins, which were reproducibly identified in all three replications, revealed that 25 proteins were statically significant differentially expressed. Among the differentially expressed proteins, Ig lambda-7 chain C region and tenascin drastically decreased with the delivery time.

Butyrate-induced differentiation of PC12 cells to chromaffin cells involves cell adhesion and induction of extracellular proteins and cell adhesion proteins

  • Heo, Jee-In;Oh, Soo-Jin;Kho, Yoon-Jung;Kim, Jeong-Hyeon;Kang, Hong-Joon;Park, Seong-Hoon;Kim, Hyun-Seok;Shin, Jong-Yeon;Lee, Sung-Young;Kim, Min-Ju;Min, Bon-Hong;Kim, Sung-Chan;Park, Jae-Bong;Kim, Jae-Bong;Lee, Jae-Yong
    • Animal cells and systems
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    • v.14 no.4
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    • pp.261-266
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    • 2010
  • PC12 cells were differentiated into the cells of chromaffin phenotype by butyrate treatment. Cells were aggregated and formed tight cell adhesion. To investigate the molecular change in this differentiation, we examined expression levels of cell adhesion proteins and extracellular proteins during butyrate induced-differentiation of PC12 cells. Integrin ${\beta}1$, integrin ${\alpha}7$, E cadherin, VCAM, collagen-I, fibronectin, desmoglein and connexin were increased during differentiation. The levels of clusterin and secreted clusterin were also increased. These increased levels of cell adhesion proteins and extracellular proteins appear to induce cell aggregation and tight cell adhesion. The levels of p21, p27 and p16 were increased probably because of differentiation-related growth arrest during differentiation. Prolonged incubation of butyrate up to 1 day was required for differentiation. Signal transduction pathways for this differentiatiom could not be identified since various inhibitors had no effect. The results showed that butyrateinduced differentiation of PC12 cells to chromaffin cells involves tight cell adhesion and induction of extracellular proteins and cell adhesion proteins.

Gene Expression Profile of Lung Cancer Cells Following Photodynamic Therapy (폐암 세포주에서 광역학 치료에 의한 유전자 발현 분석)

  • Sung, Ji Hyun;Lee, Mi-Eun;Han, Seon-Sook;Lee, Seung-Joon;Ha, Kwon-Soo;Kim, Woo Jin
    • Tuberculosis and Respiratory Diseases
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    • v.63 no.1
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    • pp.52-58
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    • 2007
  • Background: Photodynamic therapy is a viable option for lung cancer treatment, and many studies have shown that it is capable of inducing cell death in lung cancer cells. However, the precise mechanism of this cell death has not been fully elucidated. To investigate the early changes in cancer cell transcription, we treated A549 cells with the photosensitizer DH-I-180-3 and then we illuminated the cells. Methods: We investigated the gene expression profiles of the the A549 lung cancer cell line, using a DEG kit, following photodynamic therapy and we evaluated the cell viability by performing flow cytometry. We identified the genes that were significantly changed following photodynamic therapy by performing DNA sequencing. Results: The FACS data showed that the cell death of the lung cancer cells was mainly caused by necrosis. We found nine genes that were significantly changed and we identified eight of these genes. We evaluated the expression of two genes, 3-phosphoglycerate dehydrogenase and ribosomal protein S29. The expressed level of carbonic anhydrase XII, clusterin, MRP3s1 protein, complement 3, membrane cofactor protein and integrin beta 1 were decreased. Conclusion: Many of the gene products are membrane-associated proteins. The main mechanism of photodynamic therapy with using the photosensitizing agent DH-I-180-3 appears to be necrosis and this may be associated with the altered production of membrane proteins.

Parabiosis and Blood Exchange Techniques in Aging Research (개체병렬결합(parabiosis)실험모델과 혈액교환을 이용한 노화(aging)연구 분석)

  • Kyung Tae Chung
    • Journal of Life Science
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    • v.33 no.2
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    • pp.208-215
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    • 2023
  • In recent decades, the field of aging research has progressed from the genetic and cellular levels to in vivo models of blood exchange. Since genes capable of extending the lifespan in C. elegance have been reported, various potential target molecules have been discovered through genomics, proteomics, metabolomics, and transcriptomics. Accordingly, research on the interactions between target molecules has also been increasing. The parabiosis method, in which two experimental animals are surgically combined, was introduced, and a factor that could reverse the aging phenomenon was discovered using this method. The parabiosis method is used to find more accurate and effective aging-reversal factors that could exist in young blood. As more new evidence has been revealed, the parabiosis method has established a new paradigm for aging research. Moreover, a device capable of exchanging blood elaborately in laboratory animals was published in 2022 and presented new results necessary for aging reversal. Since GDF11, was reported, many other anti-aging candidates that are soluble factors in blood, such as β2m, TIMP2, VCAM1, Gpld1, and clusterin, have been discovered. In addition, mcicroglia cells and neuroinflammation have been directly proven to be aging factors. These latest research results were obtained by parabiosis, the newly designed device for plasmapheresis, and injecting young blood or conditioned blood methods. In this review, we discuss the latest research results using the device and young blood administration in old mice.

Gene Expression Analysis of Megakaryocytes Derived from Human Umbilical Cord $CD34^+$ Cells by Thrombopoietin

  • Kim, Jeong-Ah;Kim, Hyung-Lae
    • Genomics & Informatics
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    • v.3 no.1
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    • pp.8-14
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    • 2005
  • Although much is known about the molecular biology of platelets, the megakaryocytes' (MKs) molecular biology was not understood so well because of their rareness. By the cloning and characterization of thrombopoietin (TPO), which is the principal regulator of the growth and development of the MKs, researches on the MKs have been growing rapidly. To understand megakaryocytopoiesis, we investigated the gene expression profile of the MKs using oligonucleotide microarray where 10,108 unique genes were spotted. Comparing the fluorescence intensities of which ratio is $\ge$ ${\mid}2{\mid}$, 372 genes were up-regulated and 541 genes were down-regulated in MKs. For confirmatory expression, RNase protection assay (RPA) establishing abundant apoptotic gene expression was carried out. In MKs, many of the known genes, including several platelet related genes, GATA binding protein were highly expressed. Particularly, TGF beta, clusterin (complement lysis inhibitor), and thymosin beta 4 (actin-sequestering molecules) were expressed highly in MKs. As MKs specific expressed genes may regulate normal and pathologic platelet (and/or MK) functions, the transcript profiling using microarray was useful on molecular understanding of MKs,