scholarly journals Mutation of Herpesvirus Saimiri ORF51 Glycoprotein Specifically Targets Infectivity to Hepatocellular Carcinoma Cell Lines

2011 ◽  
Vol 2011 ◽  
pp. 1-14 ◽  
Author(s):  
Susan J. Turrell ◽  
Adrian Whitehouse
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Lines ◽  
Human Cell ◽  
Heparan Sulphate ◽  
Somatostatin Receptors ◽  
Cell Types ◽  
Peptide Sequence ◽  
Herpesvirus Saimiri ◽  
Wild Type Virus ◽  
Virion Protein

Herpesvirus saimiri (HVS) is a gamma herpesvirus with several properties that make it an amenable gene therapy vector; namely its large packaging capacity, its ability to persist as a nonintegrated episome, and its ability to infect numerous human cell types. We used RecA-mediated recombination to develop an HVS vector with a mutated virion protein. The heparan sulphate-binding region of HVS ORF51 was substituted for a peptide sequence which interacts with somatostatin receptors (SSTRs), overexpressed on hepatocellular carcinoma (HCC) cells. HVS mORF51 showed reduced infectivity in non-HCC human cell lines compared to wild-type virus. Strikingly, HVS mORF51 retained its ability to infect HCC cell lines efficiently. However, neutralisation assays suggest that HVS mORF51 has no enhanced binding to SSTRs. Therefore, mutation of the ORF51 glycoprotein has specifically targeted HVS to HCC cell lines by reducing the infectivity of other cell types; however, the mechanism for this targeting is unknown.

scholarly journals Expression of extracellular superoxide dismutase by human cell lines

1990 ◽  
Vol 266 (1) ◽  
pp. 213-219 ◽  
Author(s):  
S L Marklund
Keyword(s):  
Superoxide Dismutase ◽  
Cell Lines ◽  
Human Cell ◽  
Heparan Sulphate ◽  
Cell Types ◽  
Tissue Cell ◽  
Extracellular Superoxide Dismutase ◽  
Human Cell Lines ◽  
Growing Cell ◽  
Dependent Cell

Extracellular superoxide dismutase (EC-SOD) is the major SOD isoenzyme in extracellular fluids, but occurs also in tissues. The sites and characteristics of the synthesis of the enzyme are unknown. The occurrence of EC-SOD in cultures of a large panel of human cell lines was assayed by means of an e.l.i.s.a. Unlike the situation for the intracellular isoenzymes CuZn-SOD and Mn-SOD, expression of EC-SOD occurs in only a few cell types. None of the ten investigated suspension-growing cell lines produced EC-SOD. Among normal diploid anchorage-dependent cell lines, expression was found in all 25 investigated fibroblast cell lines, in the two glia-cell lines, but not in six endothelial-cell lines, two epithelial-cell lines or in two amnion-derived lines. Among neoplastic anchorage-dependent cell lines expression was found in 13 out of 29. EC-SOD was secreted into the culture medium by cell lines expressing the enzyme. The rate of EC-SOD synthesis varied by nearly 100-fold among the fibroblast lines and remained essentially constant in the individual lines during long-term culture. In the nine investigated cases, the secreted EC-SOD was of the high-heparin-affinity C type. It is suggested that tissue EC-SOD is secreted by a few well-dispersed cell types, such as fibroblasts and glia cells, to diffuse subsequently around and reversibly bind to heparan sulphate proteoglycan ligands in the glycocalyx of the surface of most tissue cell types and in the interstitial matrix.

scholarly journals Analysis of Mitochondrial DNA Polymorphisms in the Human Cell Lines HepaRG and SJCRH30

2019 ◽  
Vol 20 (13) ◽  
pp. 3245 ◽  
Author(s):  
Matthew J. Young ◽  
Anitha D. Jayaprakash ◽  
Carolyn K. J. Young
Keyword(s):  
Mitochondrial Dna ◽  
Cell Lines ◽  
Human Cell ◽  
Cell Types ◽  
Dna Polymorphisms ◽  
Reference Sequence ◽  
Sequencing Analysis ◽  
Human Cell Lines ◽  
Mtdna Sequences ◽  
Mtdna Variants

The mitochondrial DNA (mtDNA) sequences of two commonly used human cell lines, HepaRG and SJCRH30, were determined. HepaRG originates from a liver tumor obtained from a patient with hepatocarcinoma and hepatitis C while SJCRH30 originates from a rhabdomyosarcoma patient tumor. In comparison to the revised Cambridge Reference Sequence, HepaRG and SJCRH30 mtDNA each contain 14 nucleotide variations. In addition to an insertion of a cytosine at position 315 (315insC), the mtDNA sequences from both cell types share six common polymorphisms. Heteroplasmic variants were identified in both cell types and included the identification of the 315insC mtDNA variant at 42 and 75% heteroplasmy in HepaRG and SJCRH30, respectively. Additionally, a novel heteroplasmic G13633A substitution in the HepaRG ND5 gene was detected at 33%. Previously reported cancer-associated mtDNA variants T195C and T16519C were identified in SJCRH30, both at homoplasmy (100%), while HepaRG mtDNA harbors a known prostate cancer-associated T6253C substitution at near homoplasmy, 95%. Based on our sequencing analysis, HepaRG mtDNA is predicted to lie within haplogroup branch H15a1 while SJCRH30 mtDNA is predicted to localize to H27c. The catalog of polymorphisms and heteroplasmy reported here should prove useful for future investigations of mtDNA maintenance in HepaRG and SJCRH30 cell lines.

scholarly journals The Subcellular Localization of Syntaxin 17 Varies Among Different Cell Types and Is Altered in Some Malignant Cells

2005 ◽  
Vol 53 (11) ◽  
pp. 1371-1382 ◽  
Author(s):  
Qian Zhang ◽  
Jiang Li ◽  
Michael Deavers ◽  
James L. Abbruzzese ◽  
Linus Ho
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Lines ◽  
Human Cancer ◽  
Sequence Divergence ◽  
Cell Types ◽  
Human Hepatocellular Carcinoma ◽  
Cytoplasmic Localization ◽  
Malignant Cells ◽  
Human Cancer Cell Lines ◽  
Mouse Tissues

Syntaxin 17 (STX17) is a divergent member of the syntaxin family of proteins first discovered by Scheller and colleagues in a yeast two-hybrid screen designed to identify novel mammalian SNAREs (soluble N-ethylmaleimide-sensitive factor-attachment protein receptors). We recently independently identified STX17 as a novel Ras-interacting protein, but immunohistochemical studies suggested that STX17 is localized to the nucleus in normal pancreatic ductal epithelial, acinar, and islet cells in contrast to previous reports of cytoplasmic localization, albeit in other cell types. Therefore, we have conducted a more thorough survey of various human and mouse tissues to better establish the expression pattern of STX17 in different tissues and cell types. Although RT-PCR experiments demonstrate ubiquitous expression of STX17, closer examination by immunohistochemistry reveal that STX17 expression is limited to certain cell types. Furthermore, in contrast to the cytoplasmic localization previously reported in a limited number of cell types, we find that in many other cell types, syntaxin 17 can be found in the nucleus. Finally, we demonstrate that in human hepatocellular carcinoma cell lines, STX17 localization is altered relative to normal hepatocytes, although the localization of STX17 differs even between these established human cancer cell lines and fresh human hepatocellular carcinoma cells, emphasizing the caution that must be exercised in drawing conclusions from data gathered in cell lines. The sequence divergence of STX17, the unexpected nuclear localization of STX17 in many cell types, and the altered localization of STX17 in malignant cells argue for a novel function of syntaxin 17 distinct from its hypothesized role in mediating membrane fusion events.

scholarly journals SARS-CoV-2 disrupts proximal elements in the JAK-STAT pathway

2021 ◽  
Author(s):  
Da-Yuan Chen ◽  
Nazimuddin Khan ◽  
Brianna J. Close ◽  
Raghuveera K. Goel ◽  
Benjamin Blum ◽  
...  
Keyword(s):  
Cell Lines ◽  
Human Cell ◽  
Type I Interferon ◽  
Cell Types ◽  
Janus Kinase ◽  
Type I ◽  
Human Cell Lines ◽  
Interferon Signaling ◽  
Interferon Receptor ◽  
Stat Pathway

SARS-CoV-2 can infect multiple organs, including lung, intestine, kidney, heart, liver, and brain. The molecular details of how the virus navigates through diverse cellular environments and establishes replication are poorly defined. Here, we generated a panel of phenotypically diverse, SARS-CoV-2-infectable human cell lines representing different body organs and performed longitudinal survey of cellular proteins and pathways broadly affected by the virus. This revealed universal inhibition of interferon signaling across cell types following SARS-CoV-2 infection. We performed systematic analyses of the JAK-STAT pathway in a broad range of cellular systems, including immortalized cells and primary-like cardiomyocytes, and found that SARS-CoV-2 targeted the proximal pathway components, including Janus kinase 1 (JAK1), tyrosine kinase 2 (Tyk2), and the interferon receptor subunit 1 (IFNAR1), resulting in cellular desensitization to type I IFN. Detailed mechanistic investigation of IFNAR1 showed that the protein underwent ubiquitination upon SARS-CoV-2 infection. Furthermore, chemical Inhibition of JAK kinases enhanced infection of stem cell-derived cultures, indicating that the virus benefits from inhibiting the JAK-STAT pathway. These findings suggest that the suppression of interferon signaling is a mechanism widely used by the virus to evade antiviral innate immunity, and that targeting the viral mediators of immune evasion may help block virus replication in patients with COVID-19. IMPORTANCE SARS-CoV-2 can infect various organs in the human body, but the molecular interface between the virus and these organs remains unexplored. In this study, we generated a panel of highly infectable human cell lines originating from various body organs and employed these cells to identify cellular processes commonly or distinctly disrupted by SARS-CoV-2 in different cell types. One among the universally impaired processes was interferon signaling. Systematic analysis of this pathway in diverse culture systems showed that SARS-CoV-2 targets the proximal JAK-STAT pathway components, destabilizes the type I interferon receptor though ubiquitination, and consequently renders the infected cells resistant to type I interferon. These findings illuminate how SARS-CoV-2 can continue to propagate in different tissues even in the presence of a disseminated innate immune response.

scholarly journals Enhanced Cellular Uptake of H-Chain Human Ferritin Containing Gold Nanoparticles

Pharmaceutics ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 1966
Author(s):  
Italo Moglia ◽  
Margarita Santiago ◽  
Simon Guerrero ◽  
Mónica Soler ◽  
Alvaro Olivera-Nappa ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Cell Lines ◽  
Cellular Uptake ◽  
Human Cell ◽  
Animal Experiments ◽  
Cell Types ◽  
Biological Properties ◽  
Human Cell Lines ◽  
Theranostic Agents

Gold nanoparticles (AuNP) capped with biocompatible layers have functional optical, chemical, and biological properties as theranostic agents in biomedicine. The ferritin protein containing in situ synthesized AuNPs has been successfully used as an effective and completely biocompatible nanocarrier for AuNPs in human cell lines and animal experiments in vivo. Ferritin can be uptaken by different cell types through receptor-mediated endocytosis. Despite these advantages, few efforts have been made to evaluate the toxicity and cellular internalization of AuNP-containing ferritin nanocages. In this work, we study the potential of human heavy-chain (H) and light-chain (L) ferritin homopolymers as nanoreactors to synthesize AuNPs and their cytotoxicity and cellular uptake in different cell lines. The results show very low toxicity of ferritin-encapsulated AuNPs on different human cell lines and demonstrate that efficient cellular ferritin uptake depends on the specific H or L protein chains forming the ferritin protein cage and the presence or absence of metallic cargo. Cargo-devoid apoferritin is poorly internalized in all cell lines, and the highest ferritin uptake was achieved with AuNP-loaded H-ferritin homopolymers in transferrin-receptor-rich cell lines, showing more than seven times more uptake than apoferritin.

scholarly journals Exploring Chromosomal Structural Heterogeneity Across Multiple Cell Lines

2020 ◽  
Author(s):  
Ryan R. Cheng ◽  
Vinicius Contessoto ◽  
Erez Lieberman Aiden ◽  
Peter G. Wolynes ◽  
Michele Di Pierro ◽  
...  
Keyword(s):  
Cell Lines ◽  
Conformational Changes ◽  
Human Cell ◽  
Conformational Transition ◽  
Cell Types ◽  
Structural Heterogeneity ◽  
Human Cell Lines ◽  
Active Chromatin ◽  
State Process ◽  
Structural Ensembles

AbstractWe study the structural ensembles of human chromosomes across different cell types. Using computer simulations, we generate cell-specific 3D chromosomal structures and compare them to recently published chromatin structures obtained through microscopy. We demonstrate using a combination of machine learning and polymer physics simulations that epigenetic information can be used to predict the structural ensembles of multiple human cell lines. The chromosomal structures obtained in silico are quantitatively consistent with those obtained through microscopy as well as DNA-DNA proximity ligation assays. Theory predicts that chromosome structures are fluid and can only be described by an ensemble, which is consistent with the observation that chromosomes exhibit no unique fold. Nevertheless, our analysis of both structures from simulation and microscopy reveals that short segments of chromatin make transitions between a closed conformation and an open dumbbell conformation. This conformational transition appears to be consistent with a two-state process with an effective free energy cost of about four times the effective information theoretic temperature. Finally, we study the conformational changes associated with the switching of genomic compartments observed in human cell lines. Genetically identical but epigenetically distinct cell types appear to rearrange their respective structural ensembles to expose segments of transcriptionally active chromatin, belonging to the A genomic compartment, towards the surface of the chromosome, while inactive segments, belonging to the B compartment, move to the interior. The formation of genomic compartments resembles hydrophobic collapse in protein folding, with the aggregation of denser and predominantly inactive chromatin driving the positioning of active chromatin toward the surface of individual chromosomal territories.

scholarly journals High Anticancer Properties of Defatted Jatropha Curcus Seed Residue and its Active Compound, Isoamericanol A

2017 ◽  
Vol 12 (12) ◽  
pp. 1934578X1701201
Author(s):  
Ayako Katagi ◽  
Li Sui ◽  
Kazuyo Kamitori ◽  
Toshisada Suzuki ◽  
Takeshi Katayama ◽  
...  
Keyword(s):  
Cell Lines ◽  
Human Cell ◽  
Active Compound ◽  
Human Cancer ◽  
Cell Types ◽  
Biodiesel Production ◽  
Growth Effect ◽  
Biodiesel Fuel ◽  
Human Cancer Cell Lines ◽  
Mcf 7

Jatropha seeds have gained popularity as a biodiesel fuel resource. The process of biodiesel production from the seeds, however, also creates a tremendous amount of defatted residue as a byproduct. In this study, the potential for pharmaceutical usage of extracts from the defatted residue of crushed Jatropha curcus L. seed was examined. Organic and water fractions of both ethyl acetate (EtOAc) and methanol (MeOH) extracts from the seed residue exhibited an inhibitory growth effect on the human cancer cell lines hepatocellular carcinoma (HuH-7) and breast cancer (MCF-7). Later, the effect of isoamericanol A from the MeOH-organic fraction was compared across three human cell lines in search of human cell types suitable for treatment (HuH-7, MCF-7, cervical cancer (HeLa)). Our results indicate that the effect of three-day isoamericanol A treatment differs by cell type. Although effective to some extent, HeLa cell growth was most inhibited at the smallest dosage, whereas MCF-7 growth suffered the greatest inhibition at the highest dosage. Our findings reveal possibility for utilization of the Jatropha seed residue extract as well as its active compound, isoamericanol A, as practical cancer treatments for certain cell types in the future.

scholarly journals Primary osteoblasts, osteoblast precursor cells or osteoblast‐like cell lines: Which human cell types are (most) suitable for characterizing 45S5‐bioactive glass?

2019 ◽  
Vol 108 (3) ◽  
pp. 663-674 ◽  
Author(s):  
Sebastian Wilkesmann ◽  
Jörg Fellenberg ◽  
Qaisar Nawaz ◽  
Bruno Reible ◽  
Arash Moghaddam ◽  
...  
Keyword(s):  
Bioactive Glass ◽  
Cell Lines ◽  
Human Cell ◽  
Cell Types ◽  
Precursor Cells ◽  
Primary Osteoblasts

scholarly journals Initiation of mtDNA transcription is followed by pausing, and diverge across human cell types and during evolution

2016 ◽  
Author(s):  
Amit Blumberg ◽  
Edward J. Rice ◽  
Anshul Kundaje ◽  
Charles G. Danko ◽  
Dan Mishmar
Keyword(s):  
Cell Lines ◽  
Human Cell ◽  
Transcription Initiation ◽  
De Novo ◽  
Cell Types ◽  
Reference Sequence ◽  
Sequence Motif ◽  
Mtdna Sequence ◽  
Mtdna Transcription

AbstractMitochondrial DNA (mtDNA) genes are long known to be co-transcribed in polycistrones, yet it remains impossible to study nascent mtDNA transcripts quantitatively in vivo using existing tools. To this end we used deep sequencing (GRO-seq and PRO-seq) and analyzed nascent mtDNA-encoded RNA transcripts in diverse human cell lines and metazoan organisms. Surprisingly, accurate detection of human mtDNA transcription initiation sites (TIS) in the heavy and light strands revealed a novel conserved transcription pausing site near the light strand TIS, upstream to the transcription-replication transition region. This pausing site correlated with the presence of a bacterial pausing sequence motif, yet the transcription pausing index varied quantitatively among the cell lines. Analysis of non-human organisms enabled de novo mtDNA sequence assembly, as well as detection of previously unknown mtDNA TIS, pausing, and transcription termination sites with unprecedented accuracy. Whereas mammals (chimpanzee, rhesus macaque, rat, and mouse) showed a human-like mtDNA transcription pattern, the invertebrate pattern (Drosophila and C. elegans) profoundly diverged. Our approach paves the path towards in vivo, quantitative, reference sequence-free analysis of mtDNA transcription in all eukaryotes.

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