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https://www.readbyqxmd.com/read/29341059/down-regulated-reic-expression-in-lung-carcinogenesis-a-molecular-target-for-gene-therapy
#1
Lei Yang, Shuang Zhao, Pu Xia, Hua-Chuan Zheng
REIC (Reduced Expression in Immortalized Cells) gene is down-regulated in immortalized cells, compared with the normal parental counterparts. Its encoding protein could inhibit colony formation, tumor growth, and induce apoptosis. To investigate the roles of REIC expression in lung cancer, we examined REIC expression in lung cancer cells and tissues by RT-PCR or Western blot, and observed the effects of both recombinant REIC exposure and REIC overexpression on the aggressive phenotypes of lung cancer cells...
January 17, 2018: Histology and Histopathology
https://www.readbyqxmd.com/read/29340096/crispr-cas9-mediated-reversibly-immortalized-mouse-bone-marrow-stromal-stem-cells-bmscs-retain-multipotent-features-of-mesenchymal-stem-cells-mscs
#2
Xue Hu, Li Li, Xinyi Yu, Ruyi Zhang, Shujuan Yan, Zongyue Zeng, Yi Shu, Chen Zhao, Xingye Wu, Jiayan Lei, Yasha Li, Wenwen Zhang, Chao Yang, Ke Wu, Ying Wu, Liping An, Shifeng Huang, Xiaojuan Ji, Cheng Gong, Chengfu Yuan, Linghuan Zhang, Wei Liu, Bo Huang, Yixiao Feng, Bo Zhang, Rex C Haydon, Hue H Luu, Russell R Reid, Michael J Lee, Jennifer Moriatis Wolf, Zebo Yu, Tong-Chuan He
Mesenchymal stem cells (MSCs) are multipotent non-hematopoietic progenitor cells that can undergo self-renewal and differentiate into multi-lineages. Bone marrow stromal stem cells (BMSCs) represent one of the most commonly-used MSCs. In order to overcome the technical challenge of maintaining primary BMSCs in long-term culture, here we seek to establish reversibly immortalized mouse BMSCs (imBMSCs). By exploiting CRISPR/Cas9-based homology-directed-repair (HDR) mechanism, we target SV40T to mouse Rosa26 locus and efficiently immortalize mouse BMSCs (i...
December 19, 2017: Oncotarget
https://www.readbyqxmd.com/read/29339820/histone-variant-macroh2a1-plays-an-isoform-specific-role-in-suppressing-epithelial-mesenchymal-transition
#3
Dayle Q Hodge, Jihong Cui, Matthew J Gamble, Wenjun Guo
Epithelial-Mesenchymal Transition (EMT) is a biological program that plays key roles in various developmental and pathological processes. Although much work has been done on signaling pathways and transcription factors regulating EMT, the epigenetic regulation of EMT remains not well understood. Histone variants have been recognized as a key group of epigenetic regulators. Among them, macroH2A1 is involved in stem cell reprogramming and cancer progression. We postulated that macroH2A1 may play a role in EMT, a process involving reprogramming of cellular states...
January 16, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29339778/a-novel-human-muscle-cell-model-of-duchenne-muscular-dystrophy-created-by-crispr-cas9-and-evaluation-of-antisense-mediated-exon-skipping
#4
Takenori Shimo, Kana Hosoki, Yusuke Nakatsuji, Toshifumi Yokota, Satoshi Obika
Oligonucleotide-mediated splicing modulation is a promising therapeutic approach for Duchenne muscular dystrophy (DMD). Recently, eteplirsen, a phosphorodiamidate morpholino oligomer-based splice-switching oligonucleotide (SSO) targeting DMD exon 51, was approved by the U.S. Food and Drug Administration as the first antisense-based drug for DMD patients. For further exploring SSOs targeting other exons in the DMD gene, the efficacy of exon skipping and protein rescue with each SSO sequence needs evaluations in vitro...
January 16, 2018: Journal of Human Genetics
https://www.readbyqxmd.com/read/29339735/phenotype-and-functional-features-of-human-telomerase-reverse-transcriptase-immortalized-human-airway-smooth-muscle-cells-from-asthmatic-and-non-asthmatic-donors
#5
J K Burgess, A Ketheson, A Faiz, K A Limbert Rempel, B G Oliver, J P T Ward, A J Halayko
Asthma is an obstructive respiratory disease characterised by chronic inflammation with airway hyperresponsiveness. In asthmatic airways, there is an increase in airway smooth muscle (ASM) cell bulk, which differs from non-asthmatic ASM in characteristics. This study aimed to assess the usefulness of hTERT immortalisation of human ASM cells as a research tool. Specifically we compared proliferative capacity, inflammatory mediator release and extracellular matrix (ECM) production in hTERT immortalised and parent primary ASM cells from asthmatic and non-asthmatic donors...
January 16, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29337376/crispr-cas9-mediated-genome-editing-in-epstein-barr-virus-transformed-lymphoblastoid-b-cell-lines
#6
Sizun Jiang, Liang Wei Wang, Michael J Walsh, Stephen J Trudeau, Catherine Gerdt, Bo Zhao, Benjamin E Gewurz
Epstein-Barr virus (EBV) efficiently transforms primary human B cells into immortalized lymphoblastoid cell lines (LCLs), which are extensively used in human genetic, immunological and virological studies. LCLs provide unlimited sources of DNA for genetic investigation, but can be difficult to manipulate, for instance because low retroviral or lentiviral transduction frequencies hinder experiments that require co-expression of multiple components. This unit details Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 engineering for robust LCL genome editing...
January 16, 2018: Current Protocols in Molecular Biology
https://www.readbyqxmd.com/read/29337188/abrogation-of-fam20c-altered-cell-behaviors-and-bmp-signaling-of-immortalized-dental-mesenchymal-cells
#7
Chao Liu, Nan Zhou, Yu Wang, Hua Zhang, Priyam Jani, Xiaofang Wang, Yongbo Lu, Nan Li, Jing Xiao, Chunlin Qin
FAM20C mutations compromise the mineralization of skeleton and tooth in both human and mouse. Putatively, the mineralization disorder is attributed to the elevated fibroblast growth factor 23 (FGF23), which reduced the serum phosphorus by suppressing the reabsorption of phosphorus in kidney. Besides the regulation on systemic phosphorus homeostasis, FAM20C was also implicated to regulate cell behaviors and gene expression through a cell-autonomous manner. To identify the primary effects of Fam20c on dental mesenchymal cells, mouse Fam20c-deficient dental mesenchymal cells were generated by removing the floxed alleles from the immortalized mouse Fam20cf/f dental mesenchymal cells with Cre-expressing lentivirus...
January 11, 2018: Experimental Cell Research
https://www.readbyqxmd.com/read/29336891/cupua%C3%A3-u-extract-reduces-nitrosative-stress-and-modulates-inflammatory-mediators-in-the-kidneys-of-experimental-diabetes
#8
Giovana R Punaro, Deyse Y Lima, Adelson M Rodrigues, Samuel Pugliero, Margaret G Mouro, Marcelo M Rogero, Elisa M S Higa
BACKGROUND & AIMS: We have previously reported an increased nitrosative stress in the kidneys of diabetic animals, which was reduced by an antioxidant probiotic. The present study evaluated whether the extract of cupuaçu (EC), an antioxidant compound rich in polyphenols and theograndins, when administered at a dose that can be reasonably obtained through daily consumption, could delay the onset of diabetic complications in the kidney. METHODS: Mouse immortalized mesangial cells (MiMC) were placed in medium normal glucose (NG) or high glucose (HG), with or without EC (500, 100, 50 or 10 μg/mL) during 24, 48 or 72 h for analysis of viability, proliferation, nitric oxide (NO) levels and reactive oxygen species or nitrogen (ROS/RNS)...
December 26, 2017: Clinical Nutrition: Official Journal of the European Society of Parenteral and Enteral Nutrition
https://www.readbyqxmd.com/read/29335203/inhibitor-of-the-human-telomerase-reverse-trancriptase-htert-gene-promoter-induces-cell-apoptosis-via-a-mitochondrial-dependent-pathway
#9
Yuyin Li, Guojun Pan, Yue Chen, Qian Yang, Tiantian Hao, Lianbo Zhao, Long Zhao, Yusheng Cong, Aipo Diao, Peng Yu
Telomerase is aberrantly expressed in many cancers and plays an important role in the development of cellular immortality and oncogenesis, which makes it a potential cancer therapeutic target for drug discovery. Here, we constructed a firefly luciferase reporter driven by the human telomerase reverse trancriptase (hTERT) gene promoter to screen for inhibitory compounds. Compound 5c was discovered and shown to significantly inhibit the promoter activity of hTERT gene. Furthermore, five analogs of compound 5c were synthesized, and compound 8b was shown to be a more potent inhibitor of hTERT gene promoter activity and subsequent expression of hTERT mRNA and protein...
December 27, 2017: European Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/29333545/dynamic-changes-in-copper-homeostasis-and-post-transcriptional-regulation-of-atp7a-during-myogenic-differentiation
#10
Katherine E Vest, Amanda L Paskavitz, Joseph B Lee, Teresita Padilla-Benavides
Copper (Cu) is an essential metal required for activity of a number of redox active enzymes that participate in critical cellular pathways such as metabolism and cell signaling. Because it is also a toxic metal, Cu must be tightly controlled by a series of transporters and chaperone proteins that regulate Cu homeostasis. The critical nature of Cu is highlighted by the fact that mutations in Cu homeostasis genes cause pathologic conditions such as Menkes and Wilson diseases. While Cu homeostasis in highly affected tissues like the liver and brain is well understood, no study has probed the role of Cu in development of skeletal muscle, another tissue that often shows pathology in these conditions...
January 15, 2018: Metallomics: Integrated Biometal Science
https://www.readbyqxmd.com/read/29333204/aggressive-prostate-cancer-cell-nuclei-have-reduced-stiffness
#11
Zeina S Khan, Julianna M Santos, Fazle Hussain
It has been hypothesized that highly metastatic cancer cells have softer nuclei and hence would travel faster through confining environments. Our goal was to prove this untested hypothesis for prostate cells. Our nuclear creep experiments using a microfluidic channel with a narrow constriction show that stiffness of aggressive immortalized prostate cancer nuclei is significantly lower than that of immortalized normal cell nuclei and hence can be a convenient malignancy marker. Nuclear stiffness is found to be the highest for cells expressing high levels of lamin A/C but lowest for cells expressing low lamin A/C levels...
January 2018: Biomicrofluidics
https://www.readbyqxmd.com/read/29331373/establishment-and-characterization-of-five-immortalized-human-scalp-dermal-papilla-cell-lines
#12
Mi Hee Kwack, Jung Min Yang, Gong Hee Won, Moon Kyu Kim, Jung Chul Kim, Young Kwan Sung
Dermal papilla (DP) regulates the growth and cycling of hair follicles. Cultured DP cells are useful for the study of their role in relation to hair growth and regeneration. However, cultivation of human DP cells is tedious and difficult. In addition, cultured DP cells possess a relatively short replicative life span, requiring immortalized human DP cell lines. We previously established an immortalized human DP cell line, SV40T-hTERT-DPC, by introducing human telomerase reverse transcriptase (hTERT) gene into the transformed cell line, SV40T-DPC...
January 10, 2018: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29331106/kbtbd2-inhibits-the-cytotoxic-activity-of-immortalized-nk-cells-through-down-regulating-mtor-signaling-in-a-mouse-hepatocellular-carcinoma-model
#13
Kai Dai, Yabing Huang, Zubing Chen, Xiaomei Sun, Lihua Yang, Yingan Jiang
Natural killer cell (NK cell)-based immunotherapy is a promising therapeutic strategy for hepatocellular carcinoma (HCC). However, the molecular mechanisms underlying the regulation of NK cell function in the tumor sites are not completely elucidated. In this study, we identified the enhanced expression of kelch repeat and BTB (POZ) domain containing 2 (Kbtbd2) in intratumoral NK cells in a mouse HCC implantation model as a negative regulator of NK cells. To investigate this interaction,, we used a Tet-on inducible expression system to control Kbtbd2 expression in an immortalized mouse NK cell line KIL C...
January 13, 2018: European Journal of Immunology
https://www.readbyqxmd.com/read/29330927/defined-serum-free-medium-for-bioreactor-culture-of-an-immortalized-human-erythroblast-cell-line
#14
Esmond Lee, Zhong Ri Lim, Hong Yu-Chen, Bin Xia Yang, Alan Tin-Lun Lam, Allen Kuan-Liang Chen, Jaichandran Sivalingam, Shaul Reuveny, Yuin-Han Loh, Steve Kah-Weng Oh
Anticipated shortages in donated blood supply have prompted investigation of alternative approaches for in-vitro production of red blood cells (RBCs), such as expansion of conditional immortalization erythroid progenitors. However, there is a bioprocessing challenge wherein factors promoting maximal cell expansion and growth-limiting inhibitory factors are yet to be investigated. We used an erythroblast cell line (ImEry) derived from immortalizing CD71+CD235a+ erythroblast from adult peripheral blood for optimization of expansion culture conditions...
January 13, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29330675/recognizing-conserved-non-canonical-localization-patterns-of-toll-like-receptors-in-tissues-and-across-species
#15
REVIEW
Glenn Hamonic, J Alex Pasternak, Heather L Wilson
Toll-like receptors (TLR) 1, 2, 4, 5 and 6 were originally characterized as exclusively expressed on the cell surface and TLR 3, 7, 8 and 9 were said to be localized to the endosomes. However, continued research in this area shows that TLR localization may be altered across cell-types, and in response to stimulation, age or disease. Mucosal surfaces must remain tolerant to the commensal flora and thus intracellular or basal lateral localization of TLRs at mucosal surfaces may be necessary to prevent induction of an inflammatory response to commensal flora while still allowing the possibility for the receptors to prime an immune response when a pathogen has crossed the epithelial barrier...
January 12, 2018: Cell and Tissue Research
https://www.readbyqxmd.com/read/29329282/induction-of-high-mobility-group-box-1-in-vitro-and-in-vivo-by-respiratory-syncytial-virus
#16
Sara Manti, Terri J Harford, Carmelo Salpietro, Fariba Rezaee, Giovanni Piedimonte
BACKGROUND: Despite decades have passed since its discovery, accurate biomarkers of Respiratory syncytial virus (RSV) disease activity and effective therapeutic strategies are still lacking. The high mobility group box type 1 (HMGB1) protein has been proposed as a possible link between RSV and immune system, but only limited information is currently available to support this hypothesis. METHODS: Expression of HMGB1 gene and protein were analyzed by quantitative PCR, ELISA, Western blot, immunocytochemistry, and confocal microscopy in immortalized and primary human bronchial epithelial cells, as well as in rat pup lungs...
January 12, 2018: Pediatric Research
https://www.readbyqxmd.com/read/29329079/the-interactome-of-ebv-lmp1-evaluated-by-proximity-based-bioid-approach
#17
Mark A Rider, Mujeeb R Cheerathodi, Stephanie N Hurwitz, Dingani Nkosi, Lauren A Howell, Deanna C Tremblay, Xia Liu, Fanxiu Zhu, David G Meckes
Epstein-Barr virus LMP1 is an oncoprotein required for immortalizing B lymphocytes and also plays important roles in transforming non-lymphoid tissue. The discovery of LMP1 protein interactions will likely generate targets to treat EBV-associated cancers. Here, we define the broader LMP1 interactome using the recently developed BioID method. Combined with mass spectrometry, we identified over 1000 proteins across seven independent experiments with direct or indirect relationships to LMP1. Pathway analysis suggests that a significant number of the proteins identified are involved in signal transduction and protein or vesicle trafficking...
January 9, 2018: Virology
https://www.readbyqxmd.com/read/29323267/scaffold-free-generation-of-uniform-adipose-spheroids-for-metabolism-research-and-drug-discovery
#18
Aloysius J Klingelhutz, Francoise A Gourronc, Anna Chaly, David A Wadkins, Anthony J Burand, Kathleen R Markan, Sharon O Idiga, Meng Wu, Matthew J Potthoff, James A Ankrum
Adipose tissue dysfunction is critical to the development of type II diabetes and other metabolic diseases. While monolayer cell culture has been useful for studying fat biology, 2D culture often does not reflect the complexity of fat tissue. Animal models are also problematic in that they are expensive, time consuming, and may not completely recapitulate human biology because of species variation. To address these problems, we have developed a scaffold-free method to generate 3D adipose spheroids from primary or immortal human or mouse pre-adipocytes...
January 11, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29322915/comparison-alignment-and-synchronization-of-cell-line-information-between-clo-and-efo
#19
Edison Ong, Sirarat Sarntivijai, Simon Jupp, Helen Parkinson, Yongqun He
BACKGROUND: The Experimental Factor Ontology (EFO) is an application ontology driven by experimental variables including cell lines to organize and describe the diverse experimental variables and data resided in the EMBL-EBI resources. The Cell Line Ontology (CLO) is an OBO community-based ontology that contains information of immortalized cell lines and relevant experimental components. EFO integrates and extends ontologies from the bio-ontology community to drive a number of practical applications...
December 21, 2017: BMC Bioinformatics
https://www.readbyqxmd.com/read/29321319/non-structural-protein-%C3%AF-1s-is-required-for-optimal-reovirus-protein-expression
#20
Matthew B Phillips, Johnasha D Stuart, Emily J Simon, Karl W Boehme
Reovirus non-structural protein σ1s is required for the establishment of viremia and hematogenous viral dissemination. However, the function of the σ1s protein during the reovirus replication cycle is not known. In this study, we found that σ1s was required for efficient reovirus replication in SV40-immortalized endothelial cells (SVECs), mouse embryonic fibroblasts, human umbilical vein endothelial cells (HUVECs), and T84 human colonic epithelial cells. In each of these cell lines, wild type reovirus produced substantially higher viral titers than a σ1s-deficient mutant...
January 10, 2018: Journal of Virology
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