• Title/Summary/Keyword: Biological cell

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Ubiquitination of p53 is Involved in Troglitazone Induced Apoptosis in Cervical Cancer Cells

  • Chen, Hui-Min;Zhang, Ding-Guo;Wu, Jin-Xiz;Pei, Dong-Sheng;Zheng, Jun-Nian
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.5
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    • pp.2313-2318
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    • 2014
  • Peroxisome proliferator-activated receptor gamma (PPAR-${\gamma}$), a ligand-dependent nuclear transcription factor, has been found to widely exist in tumor tissues and plays an important role in affecting tumor cell growth. In this study, we investigated the effect of PPAR-${\gamma}$ on aspects of the cervical cancer malignant phenotype, such as cell proliferation and apoptosis. Cell growth assay, Western blotting, Annexin V and flow cytometry analysis consistently showed that treatment with troglitazone (TGZ, a PPAR-${\gamma}$ agonist) led to dose-dependent inhibition of cervical cancer cell growth through apoptosis, whereas T0070907 (another PPAR-${\gamma}$ antagonist) had no effect on Hela cell proliferation and apoptosis. Furthermore, we also detected the protein expression of p53, p21 and Mdm2 to explain the underlying mechanism of PPAR-${\gamma}$ on cellular apoptosis. Our work, finally, demonstrated the existence of the TGZ-PPAR-${\gamma}$-p53 signaling pathway to be a critical regulator of cell apoptosis. These results suggested that PPAR-${\gamma}$ may be a potential therapeutic target for cervical cancer.

Zeolite-Mediated Cation Exchange Enhances the Stability of mRNA during Cell-Free Protein Synthesis

  • Kim, You-Eil;Kim, Dong-Myung;Choi, Cha-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.3
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    • pp.258-261
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    • 2006
  • The addition of zeolite particles enhances the stability of mRNA molecules in a cell-free protein synthesis system. When $20{\mu}g/{\mu}L$ of zeolite (Y5.4) is added to a reaction mixture of cell-free protein synthesis, a substantial increase in protein synthesis is observed. The stabilizing effect of zeolite is most dearly observed in an in vitro translation reaction directed by purified mRNA, as opposed to a coupled transcription and translation reaction. Upon the addition of zeolite in the in vitro translation reaction, the life span of the mRNA molecules is substantially extended, leading to an 80% increase in protein synthesis. The effect of zeolite upon the mRNA stability appears be strongly related to the cation exchange (potassium to sodium) reaction. Our results demonstrate the possibility of modifying this biological process using heterogeneous, non-biological substances in a cell-free protein synthesis system.

Histochemical and Ultrastructural Studies on the Salivary Gland of a Land Snail, Nesiohelix samarangae (동양달팽이 (Nesiohelix samarangae) 타액선의 조직화학적 및 미세구조적 연구)

  • Lee, Yong-Seok;Kang, Bo-Ra;Shin, Hui-Jin;Jeong, Kye-Heon
    • The Korean Journal of Malacology
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    • v.20 no.1
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    • pp.7-16
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    • 2004
  • Histochemical and ultrastructural studies on the salivary gland and salivary duct of a land snail Nesiohelix samarangae were conducted to observe structural characteristics and function. The salivary gland consisted of one type of epithelial cell, one type of supporting cell, and six types of gland cells. Four out of six gland cell types were histochemically identified on these secretions. The one secreted acid mucopolysaccharide and the other three secreted neutral mucopolysaccharide. The salivary duct epithelium had only one type of columnar cell with microvilli on its luminal surface. The basal protoplasmic membranes of the epithelial cells were deeply infolded so many times all along the cell bases.

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CAR T Cell Immunotherapy Beyond Haematological Malignancy

  • Cedric Hupperetz;Sangjoon Lah;Hyojin Kim;Chan Hyuk Kim
    • IMMUNE NETWORK
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    • v.22 no.1
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    • pp.6.1-6.19
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    • 2022
  • Chimeric antigen receptor (CAR) T cells, which express a synthetic receptor engineered to target specific antigens, have demonstrated remarkable potential to treat haematological malignancies. However, their transition beyond haematological malignancy has so far been unsatisfactory. Here, we discuss recent challenges and improvements for CAR T cell therapy against solid tumors: Antigen heterogeneity which provides an effective escape mechanism against conventional mono-antigen-specific CAR T cells; and the immunosuppressive tumor microenvironment which provides physical and molecular barriers that respectively prevent T cell infiltration and drive T cell dysfunction and hypoproliferation. Further, we discuss the application of CAR T cells in infectious disease and autoimmunity.

Cell type-specific gene expression profiling in brain tissue: comparison between TRAP, LCM and RNA-seq

  • Kim, TaeHyun;Lim, Chae-Seok;Kaang, Bong-Kiun
    • BMB Reports
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    • v.48 no.7
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    • pp.388-394
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    • 2015
  • The brain is an organ that consists of various cell types. As our knowledge of the structure and function of the brain progresses, cell type-specific research is gaining importance. Together with advances in sequencing technology and bioinformatics, cell type-specific transcriptome studies are providing important insights into brain cell function. In this review, we discuss 3 different cell type-specific transcriptome analyses i.e., Laser Capture Microdissection (LCM), Translating Ribosome Affinity Purification (TRAP)/RiboTag, and single cell RNA-Seq, that are widely used in the field of neuroscience. [BMB Reports 2015; 48(7): 388-394]

Establishment of Baculovirus Infected Insect Cell Line Expressing Porcine Salivary Lipocalin(SAL1) Protein

  • Seo, Hee-Won;Park, Da-Young;Kim, Min-Goo;Ahn, Mi-Hyun;Ko, Ki-Narm;Ko, Ki-Sung;Ka, Hak-Hyun
    • Reproductive and Developmental Biology
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    • v.33 no.2
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    • pp.71-76
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    • 2009
  • Salivary lipocalin (SAL1) is a member of the lipocalin protein family that has a property to associate with many lipophilic molecules. The importance of SAL1 during pregnancy in pigs has been suggested by our previous study which has shown that SAL1 is expressed in the uterine endometrium in a cell type- and implantation stage-specific manner and secreted into the uterine lumen. However, function of SAL1 in the uterus during pregnancy in pigs is not known. To understand SAL1 function in the uterus during pregnancy, we generated recombinant porcine SAL1 protein in an insect cell line. Porcine SAL1 cDNA was cloned into a baculovirus expression vector using RT-PCR and total RNA from uterine endometrium on day 12 of pregnancy, and the expression vector was used to generate recombinant Bacmid containing the SAL1 gene. The recombinant Bacmid was then transfected Sf9 cell to produce recombinant baculovirus. By infecting Sf9 cell with recombinant baculovirus, we established a SAL1-expressing insect cell expression system. Immunoblot analysis confirmed SAL1 expression in the infected cells. Recombinant SAL1 produced by the Sf9 cell line will be useful for understanding physiological function of SAL1 during pregnancy in pigs.

In situ Recovery of hCTLA4Ig from Suspension Cell Cultures of Oryza sativa (형질전환 벼 현탁세포 배양에서 hCTLA4Ig의 in situ 회수)

  • Choi, Hong-Yeol;Cheon, Su-Hwan;Kwon, Jun-Young;Yun, Boreum;Hong, Seok-Mi;Kim, Sun-Dal;Kim, Dong-Il
    • KSBB Journal
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    • v.31 no.4
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    • pp.284-290
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    • 2016
  • In this research, recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced by transgenic rice cells. RAmy3D promoter was used for overcome the limitation of low expression level in transgenic plant cells, and the secretion of target protein was accomplished by signal peptide. However, the RAmy3D promoter system which can be induced only by sugar starvation causes the decrease of cell viability. As a result, cell death promotes the release of protease which degrades the target proteins. The protein stability and productivity can be significantly influenced by proteolysis activity. Therefore, development of new strategies are necessary for the in situ recovery of target proteins from cell culture media. In this study, in situ recovery was performed by various strategies. Direct addition of Protein A resin with nylon bag leads to cell death by increased shear stress and decrease in production of hCTLA4Ig by protease. Medium exchange through modified flask could recover hCTLA4Ig with high cell viability and low protease activity, on the other hand, the productivity was lower than that of control. When in situ recovery was conducted at day 7 after induction in air-lift bioreactor, 1.94-fold of hCTLA4Ig could be recovered compared to control culture without in situ recovery. Consequently, in situ recovery of hCTLA4Ig from transgenic rice cell culture could enhance productivity significantly and prevent degradation of target proteins effectively.

Enhanced Production of hCTLA4Ig by Suppressing Cell Death in Transgenic Rice Cell Suspension Cultures (형질전환 벼 현탁세포 배양에서 세포 사멸 억제를 통한 hCTLA4Ig 생산성 증대)

  • Kim, Myong-Sik;Nam, Hyung-Jin;Kim, Min-Sub;Kwon, Jun-Young;Kim, Dong-Il
    • KSBB Journal
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    • v.28 no.4
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    • pp.260-268
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    • 2013
  • Transgenic plant cell cultures are an attractive expression system for the production of industrial and pharmaceutical proteins because of their advantages in safety and low production cost. Human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced and secreted when sugar was depleted in culture medium by transgenic rice cell lines (Oryza sativa L.) using RAmy3D promoter. Due to the production of the target protein by sugar depletion, concomitant occurrence of cell death is inevitable. For that reason, inhibition of cell death for enhancing productivity was necessary for the production period without energy sources. Supplementation of 0.1 mM sodium nitroprusside improved cell viability by 1.4-fold and maximum hCTLA4Ig production by 1.3-fold compared to those of control. Addition of 1 and 10 mM glutathione, N-acetylcysteine (NAC), and nicotinamide inhibited apoptotic-like programmed cell death by decreasing the activity of reactive oxygen species. Production hCTLA4Ig was enhanced 1.4-, 1.25-, and 1.15-fold with 10 mM NAC, 1 mM NAC, and 1 mM glutathione, respectively. In addition, it was found that the supplementation of NAC enhanced the cell viability.

Partial Characterization of the Pathogenic Factors Related to Chlamydia trachomatis Invasion of the McCoy Cell Membrane

  • Yeo, Myeng-Gu;Kim, Young-Ju;Park, Yeal
    • Journal of Microbiology
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    • v.41 no.2
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    • pp.137-143
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    • 2003
  • The present study was performed to identify pathogenic factors of Chlamydia trachomatis, which invade the host cell membrane. We prepared monoclonal antibody against C. trachomatis and searched for pathogenic factors using this antibody, and subsequently identified the surface components of the elementary body of C. trachomatis, i.e., major outer membrane protein (MOMP), lipopolysaccharide (LPS), and two other surface exposure proteins. These proteins are believed to be important in the pathogenesis of host cell chlamydial infection. Additionally, to identify factors related to the host cell and C. trachomatis, we prepared C. trachomatis infected and non-infected McCoy cell extracts, and reacted these with anti-chlamydial LPS monoclonal antibody. We found that anti-chlamydial LPS monoclonal antibody reacted with a 116 kDa proteinaceous McCoy cell membrane component.

Functional annotation of lung cancer-associated genetic variants by cell type-specific epigenome and long-range chromatin interactome

  • Lee, Andrew J.;Jung, Inkyung
    • Genomics & Informatics
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    • v.19 no.1
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    • pp.3.1-3.12
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    • 2021
  • Functional interpretation of noncoding genetic variants associated with complex human diseases and traits remains a challenge. In an effort to enhance our understanding of common germline variants associated with lung cancer, we categorize regulatory elements based on eight major cell types of human lung tissue. Our results show that 21.68% of lung cancer-associated risk variants are linked to noncoding regulatory elements, nearly half of which are cell type-specific. Integrative analysis of high-resolution long-range chromatin interactome maps and single-cell RNA-sequencing data of lung tumors uncovers number of putative target genes of these variants and functionally relevant cell types, which display a potential biological link to cancer susceptibility. The present study greatly expands the scope of functional annotation of lung cancer-associated genetic risk factors and dictates probable cell types involved in lung carcinogenesis.