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https://www.readbyqxmd.com/read/28447987/live-cell-imaging-and-3d-analysis-of-angiotensin-receptor-type-1a-trafficking-in-transfected-human-embryonic-kidney-cells-using-confocal-microscopy
#1
Parnika Kadam, Ryan McAllister, Jeffrey S Urbach, Kathryn Sandberg, Susette C Mueller
Live-cell imaging is used to simultaneously capture time-lapse images of angiotensin type 1a receptors (AT1aR) and intracellular compartments in transfected human embryonic kidney-293 (HEK) cells following stimulation with angiotensin II (Ang II). HEK cells are transiently transfected with plasmid DNA containing AT1aR tagged with enhanced green fluorescent protein (EGFP). Lysosomes are identified with a red fluorescent dye. Live-cell images are captured on a laser scanning confocal microscope after Ang II stimulation and analyzed by software in three dimensions (3D, voxels) over time...
March 27, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28415797/rab7-gtpase-controls-lipid-metabolic-signaling-in-myeloid-derived-suppressor-cells
#2
Xinchun Ding, Wenjing Zhang, Ting Zhao, Cong Yan, Hong Du
Lysosomal acid lipase (LAL) is a critical neutral lipid metabolic enzyme that regulates metabolic reprogramming in myeloid-derived suppressor cells (MDSCs) through over-activation of mammalian target of rapamycin (mTOR). Affymetrix GeneChip microarray analysis of MDSCs from LAL deficient mouse (lal-/-) revealed upregulation of Rab7 GTPase protein, which belongs to a superfamily of small-molecular-weight GTPase known to regulate intracellular membrane trafficking from early to late endosomes and lysosomes. Here, the physical protein-protein interaction between Rab7 GTPase and mTOR has been detected by co-immunoprecipitation in the cell extract of wild type HD1A and lal-/- MDSC-like HD1B myeloid cell lines...
March 16, 2017: Oncotarget
https://www.readbyqxmd.com/read/28383524/cytosolic-co-delivery-of-mirna-34a-and-docetaxel-with-core-shell-nanocarriers-via-caveolae-mediated-pathway-for-the-treatment-of-metastatic-breast-cancer
#3
Li Zhang, Xin Yang, Yaqi Lv, Xiaofei Xin, Chao Qin, Xiaopeng Han, Lei Yang, Wei He, Lifang Yin
Co-delivery of microRNAs and chemotherapeutic drugs into tumor cells is an attractive strategy for synergetic breast cancer therapy due to their complementary mechanisms. In this work, a core-shell nanocarrier coated by cationic albumin was developed to simultaneously deliver miRNA-34a and docetaxel (DTX) into breast cancer cells for improved therapeutic effect. The co-delivery nanocarriers showed a spherical morphology with an average particle size of 183.9 nm, and they efficiently protected miRNA-34a from degradation by RNase and serum...
April 6, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28298487/tgf-%C3%AE-triggers-rapid-fibrillogenesis-via-a-novel-t%C3%AE-rii-dependent-fibronectin-trafficking-mechanism
#4
Archana Varadaraj, Laura M Jenkins, Priyanka Singh, Anindya Chanda, John Snider, N Y Lee, Ayelet R Amsalem-Zafran, Marcelo Ehrlich, Yoav I Henis, Karthikeyan Mythreye
Fibronectin (FN) is a critical regulator of extra cellular matrix (ECM) remodeling through its availability and stepwise polymerization for fibrillogenesis. Availability of FN is regulated by its synthesis and turnover, while fibrillogenesis is a multistep integrin dependent process essential for cell migration, proliferation and tissue function. Transforming growth factor beta (TGF-β) is an established regulator of ECM remodeling through transcriptional control of ECM proteins. Here, we show that TGF-β, through increased FN trafficking in a transcription and SMAD independent manner, is a direct and rapid inducer of fibrillogenesis required for TGF-β induced cell migration...
March 15, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/28270132/effects-of-the-lysosomal-destabilizing-drug-siramesine-on-glioblastoma-in-vitro-and-in-vivo
#5
Stine S Jensen, Stine A Petterson, Bo Halle, Charlotte Aaberg-Jessen, Bjarne W Kristensen
BACKGROUND: Glioblastoma is the most frequent and most malignant brain tumor with the patients having a median survival of only 14.6 months. Although glioblastoma patients are treated with surgery, radiation and chemotherapy recurrence is inevitable. A stem-like population of radio- and chemoresistant brain tumor-initiating cells combined with the invasive properties of the tumors is believed to be critical for treatment resistance. In the present study, the aim was to investigate the effect of a novel therapeutic strategy using the lysosomotropic detergent siramesine on glioblastomas...
March 7, 2017: BMC Cancer
https://www.readbyqxmd.com/read/28248633/evaluation-of-smartflare-probe-applicability-for-verification-of-rnas-in-early-equine-conceptuses-equine-dermal-fibroblast-cells-and-trophoblastic-vesicles
#6
S Budik, W Tschulenk, S Kummer, I Walter, C Aurich
Live cell RNA imaging has become an important tool for studying RNA localisation, dynamics and regulation in cultured cells. Limited information is available using these methods in more complex biological systems, such as conceptuses at different developmental stages. So far most of the approaches rely on microinjection of synthetic constructs into oocytes during or before fertilisation. Recently, a new generation of RNA-specific probes has been developed, the so named SmartFlare probes (Merck Millipore). These consist of a central 15-nm gold particle with target-specific DNAs immobilised on its surface...
March 2, 2017: Reproduction, Fertility, and Development
https://www.readbyqxmd.com/read/28245476/laptm4b-predicts-axillary-lymph-node-metastasis-in-breast-cancer-and-promotes-breast-cancer-cell-aggressiveness-in-vitro
#7
Min Xiao, Shanshan Yang, FanLing Meng, Yu Qin, Yue Yang, Shusheng Jia, XiuMing Cai, Changli Li, Yuanxi Huang, Xiaoming Ning
PURPOSE: Lysosome-associated protein transmembrane-4 beta (LAPTM4B) is associated with the prognosis of several human malignancies. In this study, the role of LAPTM4B in the metastatic potential of breast cancer (BC) and its underlying molecular mechanisms were investigated. METHODS: The relationship between LAPTM4B expression and axillary lymph node metastasis was determined in 291 BC specimens by immunohistochemistry. The expression of LAPTM4B in paired BC cells was overexpressed and inhibited to analyse the role of LAPTM4B in the aggressiveness of BC...
February 27, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28218663/advanced-glycation-end-products-inhibit-the-proliferation-of-human-umbilical-vein-endothelial-cells-by-inhibiting-cathepsin-d
#8
Yuan Li, Ye Chang, Ning Ye, Dongxue Dai, Yintao Chen, Naijin Zhang, Guozhe Sun, Yingxian Sun
We aimed to investigate the effect of advanced glycation end products (AGEs) on the proliferation and migration ability of human umbilical vein endothelial cells (HUVECs). Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay, real-time cell analyzer and 5-Ethynyl-2'-deoxyuridine (EdU) staining. Cell migration was detected by wound-healing and transwell assay. AGEs significantly inhibited the proliferation and migration of HUVECs in a time-and dose-dependent way. Western blotting revealed that AGEs dramatically increased the expression of microtubule-associated protein 1 light chain 3 (LC3) II/I and p62...
February 17, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28117540/cathepsin-k-modulates-invasion-migration-and-adhesion-of-oral-squamous-cell-carcinomas-in%C3%A2-vitro
#9
K Yamashita, M Iwatake, K Okamoto, S-I Yamada, M Umeda, T Tsukuba
OBJECTIVE: Cathepsin K was initially discovered as an osteoclast-specific cysteine proteinase, but the enzyme is also expressed in various cancers including oral squamous cell carcinomas. This study aimed to clarify the function of cathepsin K in oral squamous cell carcinomas. MATERIALS AND METHODS: Expression levels of cathepsin K were examined in six types of cell carcinomas. Carcinomas overexpressing cathepsin K were constructed. Effects of cathepsin K overexpression and treatment with odanacatib, a specific cathepsin K inhibitor, on cell invasion, migration and adhesion were analysed...
January 24, 2017: Oral Diseases
https://www.readbyqxmd.com/read/28115068/the-role-of-oxidative-stress-in-dna-damage-in-pancreatic-%C3%AE-cells-induced-by-di-2-ethylhexyl-phthalate
#10
Yan She, Liping Jiang, Liangliang Zheng, Heming Zuo, Min Chen, Xiance Sun, Qiujuan Li, Chengyan Geng, Guang Yang, Lijie Jiang, Xiaofang Liu
Di(2-ethyhexyl) phthalate (DEHP) is commonly used as a plasticizer, which loosely binds to plastic materials and easily leaches out of these products and enters into the environment. Exposure to DEHP can impair pancreatic beta cells (INS-1 cells)function, which is associated with Insulin Resistance (IR) and type 2 diabetes. However, the mechanism of how DEHP leads to Insulin Resistance is unknown. Our results showed that the cell viability of INS-1 cells exposed to DEHP (0-1600 μM) were decreased in a concentration-dependent manner...
March 1, 2017: Chemico-biological Interactions
https://www.readbyqxmd.com/read/28106765/quantitative-proteomic-profiling-of-tachyplesin-i-targets-in-u251-gliomaspheres
#11
Xuan Li, Jianguo Dai, Yongjun Tang, Lulu Li, Gang Jin
Tachyplesin I is a cationic peptide isolated from hemocytes of the horseshoe crab and its anti-tumor activity has been demonstrated in several tumor cells. However, there is limited information providing the global effects and mechanisms of tachyplesin I on glioblastoma multiforme (GBM). Here, by using two complementary proteomic strategies (2D-DIGE and dimethyl isotope labeling-based shotgun proteomics), we explored the effect of tachyplesin I on the proteome of gliomaspheres, a three-dimensional growth model formed by a GBM cell line U251...
January 18, 2017: Marine Drugs
https://www.readbyqxmd.com/read/28068557/cordycepin-inhibits-migration-of-human-glioblastoma-cells-by-affecting-lysosomal-degradation-and-protein-phosphatase-activation
#12
Dueng-Yuan Hueng, Ching-Hsuan Hsieh, Yu-Chen Cheng, Wen-Chiuan Tsai, Ying Chen
Cordycepin, a nucleoside-derivative-isolated form Cordyceps militaris, has been reported to suppress tumor cell proliferation and cause apoptosis. This study investigates the effect of cordycepin on the migration of human glioblastoma cells. Cordycepin suppressed the migration of the human glioblastoma cell lines U87MG and LN229 in transwell and wound healing assays. Cordycepin decreased protein expression of integrin α1, focal adhesion kinase (FAK), p-FAK, paxillin and p-paxillin. The lysosomal inhibitor NH4Cl blocked the ability of cordycepin to inhibit focal adhesion protein expression and glioma cell migration...
December 29, 2016: Journal of Nutritional Biochemistry
https://www.readbyqxmd.com/read/28028177/autophagy-mediated-regulation-of-bace1-protein-trafficking-and-degradation
#13
Tuancheng Feng, Prasad Tammineni, Chanchal Agrawal, Yu Young Jeong, Qian Cai
β-Site amyloid precursor protein (APP) cleaving enzyme 1 (BACE1) is the major neuronal β-secretase for amyloid-β generation and is degraded in lysosomes. The autophagy-lysosomal system plays a key role in the maintenance of cellular homeostasis in neurons. Recent studies established that nascent autophagosomes in distal axons move predominantly in the retrograde direction toward the soma, where mature lysosomes are mainly located. However, it remains unknown whether autophagy plays a critical role in regulation of BACE1 trafficking and degradation...
February 3, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27989700/cathepsin-l-is-involved-in-x-ray-induced-invasion-and-migration-of-human-glioma-u251-cells
#14
Yajie Xiong, Wenjun Ji, Yao Fei, Yifan Zhao, Long Wang, Wenjuan Wang, Meilin Han, Caihong Tan, Xifeng Fei, Qiang Huang, Zhongqin Liang
An important therapeutic method of glioblastoma, the most common primary brain tumor, is radiotherapy. However, several studies reported recently that radiation could also promote the invasion and migration of malignant tumor. Herein, we have identified that a significant increase of migration and invasiveness of human glioma U251 cells undergoing X-ray was observed compared to controls, accompanied by the increase of cathepsin L (CTSL), which is a lysosomal cysteine protease overexpressed and secreted by tumor cells...
January 2017: Cellular Signalling
https://www.readbyqxmd.com/read/27980069/multiple-routes-of-endocytic-internalization-of-pdgfr%C3%AE-contribute-to-pdgf-induced-stat3-signaling
#15
Kamil Jastrzębski, Daria Zdżalik-Bielecka, Agnieszka Mamińska, Yannis Kalaidzidis, Carina Hellberg, Marta Miaczynska
Platelet-derived growth factor receptor β (PDGFRβ) is a receptor tyrosine kinase which upon activation by PDGF-BB stimulates cell proliferation, migration and angiogenesis. Ligand binding induces intracellular signaling cascades but also internalization of the receptor, eventually resulting in its lysosomal degradation. However, endocytic trafficking of receptors often modulates their downstream signaling. We previously reported that internalization of PDGFRβ occurs via dynamin-dependent and -independent pathways but their further molecular determinants remained unknown...
February 1, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/27974833/tissue-extract-fractions-from-starfish-undergoing-regeneration-promote-wound-healing-and-lower-jaw-blastema-regeneration-of-zebrafish
#16
Yancen Dai, Nagarajan Prithiviraj, Jianhong Gan, Xin A Zhang, Jizhou Yan
Natural bioactive materials provide an excellent pool of molecules for regenerative therapy. In the present study, we amputate portions of the arms of Archaster typicus starfish, extract and separate the active biomaterials, and compare the effects of each fraction on in vitro wound healing and in vivo lower jaw regeneration of zebrafish. Compared with crude extract, normal hexane fractions (NHFs) have a remarkable effect on cellular proliferation and collective migration, and exhibit fibroblast-like morphology, while methanol-water fractions (MWFs) increase cell size, cell-cell adhesion, and cell death...
December 15, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27974566/cd63-regulates-epstein-barr-virus-lmp1-exosomal-packaging-enhancement-of-vesicle-production-and-noncanonical-nf-%C3%AE%C2%BAb-signaling
#17
Stephanie N Hurwitz, Dingani Nkosi, Meghan M Conlon, Sara B York, Xia Liu, Deanna C Tremblay, David G Meckes
Latent membrane protein 1 (LMP1) is an Epstein-Barr virus (EBV)-encoded oncoprotein that is packaged into small extracellular vesicles (EVs) called exosomes. Trafficking of LMP1 into multivesicular bodies (MVBs) alters the content and function of exosomes. LMP1-modified exosomes enhance the growth, migration, and invasion of malignant cells, demonstrating the capacity to manipulate the tumor microenvironment and enhance the progression of EBV-associated cancers. Despite the growing evidence surrounding the significance of LMP1-modified exosomes in cancer, very little is understood about the mechanisms that orchestrate LMP1 incorporation into these vesicles...
March 1, 2017: Journal of Virology
https://www.readbyqxmd.com/read/27942972/the-influenza-a-virus-matrix-protein-2-undergoes-retrograde-transport-from-the-endoplasmic-reticulum-into-the-cytoplasm-and-bypasses-cytoplasmic-proteasomal-degradation
#18
Sanchari Bhowmick, Chandrani Chakravarty, Shanmugaapriya Sellathamby, Sunil K Lal
The matrix protein 2 (M2) is a spliced product of segment 7 genome of influenza A virus. Previous studies indicate its role in uncoating of the viral ribonucleoprotein complex during viral entry and in membrane scission while budding. Despite its crucial role in the viral life cycle, little is known about its subcellular distribution and dynamics. In this study, we have shown that the M2 protein is translocated from the membrane to the cytoplasm by a retrograde route via endosomes and the Golgi network. It utilizes retromer cargo while moving from the endosome to the trans-Golgi network and prevents endosome fusion with the lysosome...
April 2017: Archives of Virology
https://www.readbyqxmd.com/read/27922670/downregulation-of-laptm5-suppresses-cell-proliferation-and-viability-inducing-cell-cycle-arrest-at-g0-g1-phase-of-bladder-cancer-cells
#19
Liang Chen, Gang Wang, Yi Luo, Yongzhi Wang, Conghua Xie, Wei Jiang, Yu Xiao, Guofeng Qian, Xinghuan Wang
Our transcriptome analysis revealed in bladder cancer (BCa) tissues a significant induction of lysosomal-associated multispanning membrane protein 5 (LAPTM5), a lysosomal membrane protein preferentially expressing in immune cells and hematopoietic cells. Transportation of LAPTM5 from Golgi to lysosome could be inhibited by deficiency of Nedd4, a key member of E3 ubiquitin ligase family overexpressing in invasive BCa and promoting its progression. Therefore, we hypothesize that LAPTM5 may be closely correlated with BCa tumorigenesis...
January 2017: International Journal of Oncology
https://www.readbyqxmd.com/read/27897122/bioinformatics-analysis-of-chicken-mirnas-associated-with-monocyte-to-macrophage-differentiation-and-subsequent-ifng-stimulated-activation
#20
Kristopher Jl Irizarry, Adam Chan, Derek Kettle, Steven Kezian, Dominic Ma, Louis Palacios, Qingshun Li, Calvin L Keeler, Yvonne Drechsler
The goal of this project was to characterize the molecular and cellular roles of various gene targets regulated by 8 miRNAs in differentiating macrophages. Among a number of miRNAs that are found to be expressed in avian macrophages, we focused on eight specific miRNAs (miR-1618, miR-1586, miR-1633, miR-1627, miR-1646, miR-1649, miR-1610, miR-1647) associated with macrophage activation through Wnt signaling, ubiquitination, PPAR mediated macrophage function, vesicle mediated cytokine trafficking, and WD40 domain proteins in macrophage differentiation...
November 29, 2016: MicroRNA
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