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Start Over Author "Cao, Wanlu" Topic cell proliferation
4 results on '"Cao, Wanlu"'

1. Dynamics of Proliferative and Quiescent Stem Cells in Liver Homeostasis and Injury.

Author

Cao W, Chen K, Bolkestein M, Yin Y, Verstegen MMA, Bijvelds MJC, Wang W, Tuysuz N, Ten Berge D, Sprengers D, Metselaar HJ, van der Laan LJW, Kwekkeboom J, Smits R, Peppelenbosch MP, and Pan Q

Subjects
Animals, Bile Ducts metabolism, Bile Ducts pathology, Carbon Tetrachloride, Cell Differentiation, Cell Lineage, Cells, Cultured, Chemical and Drug Induced Liver Injury genetics, Chemical and Drug Induced Liver Injury metabolism, Diphtheria Toxin genetics, Diphtheria Toxin metabolism, Disease Models, Animal, Gene Expression Regulation, Developmental, Genotype, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, Hepatocytes metabolism, Hepatocytes pathology, Liver metabolism, Mice, Transgenic, Phenotype, Promoter Regions, Genetic, RNA, Untranslated genetics, Receptors, G-Protein-Coupled genetics, Stem Cell Niche, Stem Cells metabolism, Time Factors, Cell Proliferation, Cellular Senescence, Chemical and Drug Induced Liver Injury pathology, Liver pathology, Liver Regeneration, Stem Cells pathology
Abstract

Background & Aims: Adult liver stem cells are usually maintained in a quiescent/slow-cycling state. However, a proliferative population, marked by leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5), was recently identified as an important liver stem cell population. We aimed to investigate the dynamics and functions of proliferative and quiescent stem cells in healthy and injured livers., Methods: We studied LGR5-positive stem cells using diphtheria toxin receptor and green fluorescent protein (GFP) knock-in mice. In these mice, LGR5-positive cells specifically coexpress diphtheria toxin receptor and the GFP reporter. Lineage-tracing experiments were performed in mice in which LGR5-positive stem cells and their daughter cells expressed a yellow fluorescent protein/mTmG reporter. Slow-cycling stem cells were investigated using GFP-based, Tet-on controlled transgenic mice. We studied the dynamics of both stem cell populations during liver homeostasis and injury induced by carbon tetrachloride. Stem cells were isolated from mouse liver and organoid formation assays were performed. We analyzed hepatocyte and cholangiocyte lineage differentiation in cultured organoids., Results: We did not detect LGR5-expressing stem cells in livers of mice at any stage of a lifespan, but only following liver injury induced by carbon tetrachloride. In the liver stem cell niche, where the proliferating LGR5 + cells are located, we identified a quiescent/slow-cycling cell population, called label-retaining cells (LRCs). These cells were present in the homeostatic liver, capable of retaining the GFP label over 1 year, and expressed a panel of progenitor/stem cell markers. Isolated single LRCs were capable of forming organoids that could be carried in culture, expanded for months, and differentiated into hepatocyte and cholangiocyte lineages in vitro, demonstrating their bona fide stem cell properties. More interestingly, LRCs responded to liver injury and gave rise to LGR5-expressing stem cells, as well as other potential progenitor/stem cell populations, including SOX9- and CD44-positive cells., Conclusions: Proliferative LGR5 cells are an intermediate stem cell population in the liver that emerge only during tissue injury. In contrast, LRCs are quiescent stem cells that are present in homeostatic liver, respond to tissue injury, and can give rise to LGR5 stem cells, as well as SOX9- and CD44-positive cells., (Copyright © 2017 AGA Institute. Published by Elsevier Inc. All rights reserved.)

Published
2017
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2. Periplasmic expression optimization of VEGFR2 D3 adopting response surface methodology: Antiangiogenic activity study

Author

Cao Wanlu, Haixin Li, Min Wang, Juan Zhang, Daojuan Li, Zhiguo Chen, and Desmond Omane Acheampong

Subjects
Vascular Endothelial Growth Factor A, Angiogenesis, Neovascularization, Physiologic, Angiogenesis Inhibitors, Chick Embryo, Biology, Chorioallantoic Membrane, chemistry.chemical_compound, Osmotic Pressure, In vivo, Cell surface receptor, Escherichia coli, Human Umbilical Vein Endothelial Cells, Animals, Humans, Receptor, Cell Proliferation, Periplasmic space, Vascular Endothelial Growth Factor Receptor-2, Recombinant Proteins, Protein Structure, Tertiary, Vascular endothelial growth factor, Biochemistry, chemistry, Periplasm, Tumor promotion, Signal transduction, Signal Transduction, Biotechnology
Abstract

Vascular endothelial growth factor (VEGF) is one of the most significant mediators of angiogenesis, which interacts with a specific membrane receptor: VEGF receptor 2 (VEGFR2). Studies elsewhere have shown that, a VEGF-blocker can regulate several vital processes of tumor promotion. However, there is no literature evidence of investigation on antiangiogenic ability of single domain 3 of VEGFR-2 (VEGFR2 D3), as the key domain in signal transduction of VEGF. In this article, we aimed at developing an efficient method for producing soluble form of this receptor as therapeutic applications. The optimization of the production of soluble VEGFR2 D3 in Escherichia coli was firstly done by testing the periplasmic expression in different expression systems using three osmotic shock methods. To enhance the yield, vital factors were selected from nine factors by Plackett-Burman design and the level of each viral factor was optimized via a response surface methodology based central composite design. After purification and identification of the protein, the bioactivity assays: quantitative ELISA, VEGF-induced proliferation and in vivo chick chorioallantoic membrane assay were employed in our study. The outcome showed that, E. coli Rosetta-gami (DE3)/pET22b-VEGFR2 D3 was the most effective expression system. Furthermore, the inducing time, peptone and glycerol concentration affected the periplasmic expression of VEGFR2 D3 significantly. The corresponding level was also optimized. The bioactivity assay studies showed VEGFR2 D3 could suppress both VEGF stimulated cell proliferation in vitro and neovascularization in vivo. We have therefore provided a novel antiangiogenic drug candidate relating to VEGF-VEGFR2 pathway.

Published
2013

3. Action and clinical significance of CCAAT/enhancer-binding protein delta in hepatocellular carcinoma.

Author

Liu, Pengyu, Cao, Wanlu, Ma, Buyun, Li, Meng, Chen, Kan, Sideras, Kostandinos, Duitman, Jan-Willem, Sprengers, Dave, Tran, T C Khe, Ijzermans, Jan N M, Biermann, Katharina, Verheij, Joanne, Spek, C Arnold, Kwekkeboom, Jaap, Pan, Qiuwei, and Peppelenbosch, Maikel P

Subjects
*CARRIER proteins, *PROTEIN binding, *HEPATOCELLULAR carcinoma
Abstract

CCAAT/enhancer-binding protein delta (CEBPD) is associated with the regulation of apoptosis and cell proliferation and is a candidate tumor suppressor gene. Here, we investigated its role in hepatocellular carcinoma (HCC). We observe that CEBPD mRNA expression is significantly downregulated in HCC tumors as compared with adjacent tissues. Protein levels of CEBPD are also lower in tumors relative to adjacent tissues. Reduced expression of CEBPD in the tumor correlates with worse clinical outcome. In both Huh7 and HepG2 cells, shRNA-mediated CEBPD knockdown significantly reduces cell proliferation, single cell colony formation and arrests cells in the G0/G1 phase. Subcutaneous xenografting of Huh7 in nude mice show that CEBPD knockdown results in smaller tumors. Gene expression analysis shows that CEBPD modulates interleukin-1 signaling. We conclude that CEBPD expression uncouples cancer compartment expansion and clinical outcome in HCC, potentially by modulating interleukin-1 signaling. Thus, although our results support the notion that CEBPD acts as a tumor suppressor in HCC, its action does not involve impairing compartment expansion per se but more likely acts through improving anticancer immunity. [ABSTRACT FROM AUTHOR]

Published
2019
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4. The antiangiogenic activity of a soluble fragment of the VEGFR extracellular domain

Author

Daojuan Li, Cao Wanlu, Min Wang, Haizhen Jin, Desmond Omane Acheampong, Zhiguo Chen, Juan Zhang, and Haixin Li

Subjects
Vascular Endothelial Growth Factor A, Angiogenesis, Angiogenesis Inhibitors, Transfection, Chorioallantoic Membrane, Neovascularization, chemistry.chemical_compound, Epidermal growth factor, Cell Movement, medicine, Humans, MTT assay, Receptor, Cells, Cultured, Cell Proliferation, Pharmacology, General Medicine, Molecular biology, Vascular Endothelial Growth Factor Receptor-2, Peptide Fragments, Protein Structure, Tertiary, Vascular endothelial growth factor, Dissociation constant, Chorioallantoic membrane, chemistry, Solubility, Endothelium, Vascular, medicine.symptom
Abstract

Vascular endothelial growth factor (VEGF) is a key regulator of pathological angiogenesis and vascular permeability and overexpressed by most solid tumors. VEGF receptor-2 (VEGFR-2 or kinase-insert domain-containing receptor as it is called in human, KDR) is a specific receptor of VEGF with a high binding affinity. A solube recombinant extracellular domain 1-3 of human VEGFR-2 (rKDR1-3) was expressed in Escherichia coli ( E. Coli) and purified from the bacterial periplasmic extracts by immobilized metal affinity chromatography and anion exchange chromatography to inhibit the VEGF-induced angiogenesis. A surface plasmon resonance (SPR) technology was adopted to analyze the affinity and kinetics constant between rKDR1-3 and VEGF 165. Under the given experimental conditions, the association rate constant Ka was 1.06 × 10 5 M −1 S −1 , the dissociation rate Kd was 6.09 × 10 −3 S −1 , the dissociation constant KD was 5.74 × 10 −8 M. The effect of rKDR1-3 on VEGF-induced endothelial cell proliferation was studied using MTT assay, scratch-wound healing assay and chorioallantoic membrane (CAM) assay. The results showed that rKDR1-3 could inhibit neovascularization and serve as a useful drug candidate in research, diagnostics and therapy of cancer.

Published
2013

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