Cell Adhesive Interactions Research Articles

Abstract 202: Depletion of PTP4A3 phosphatase disrupts colorectal cancer cell adhesion and extracellular matrix interactions

Abstract Colorectal cancer (CRC) is the third most commonly diagnosed cancer in the United States and the world and the 5-year survival rate in the United States of ∼60% highlights the need for better treatment options. The central challenge in the design and development of drugs to treat CRC has been the validation of novel drug targets and the generation of new therapies aimed at those targets. The goal of this project is to better understand the role of one such potential CRC molecular target. Protein Tyrosine Phosphatase 4A3 (PTP4A3) is currently among the most consistently up-regulated proteins seen in colorectal tumors but it suffers from an incomplete understanding of its substrate(s) and of its participation in tumor formation and progression. To clarify how PTP4A3 expression impacts cell behavior, we isolated primary colon tumor epithelial cells from Ptp4a3fl/fl mice. PTP4A3 was deleted in vitro by adenoviral infection of Ptp4a3fl/fl cells with Cre recombinase accompanied by a green fluorescent protein (GFP) marker. As a control, the Ptp4a3fl/fl cells were infected with GFP alone. Virally infected cells were selected by fluorescence activated cell sorting. In the sorted cells, the expression of Cre resulted in a complete lack of PTP4A3 protein by western blotting and undetectable Ptp4a3 mRNA as determine by quantitative RT-PCR. Analysis of these genetically similar cells revealed that Ptp4a3−/− tumor cells had significantly reduced colony formation and migration as compared to Ptp4a3fl/fl cells. Notably, the growth rate of Ptp4a3fl/fl and Ptp4a3−/− cells was similar when cultured on plastic in a 2D monolayer. In contrast, Ptp4a3fl/fl spheroids grew significantly faster than the Ptp4a3−/− cells when cultured in Matrigel. Moreover, while the Ptp4a3fl/fl cells were able to form cohesive spheroids when cultured in cell repellent plates, there was a striking inability of the Ptp4a3−/− cells to form these 3D structures. In addition, treatment of the Ptp4a3fl/fl cells with a potent and selective PTP4A3 phosphatase inhibitor, thienopyridone, phenocopied the appearance of the Ptp4a3−/− cells grown in cell repellent plates. These results suggest PTP4A3 expression-dependent changes in tumor cell behavior may be rooted in a disruption in the adhesion and extracellular matrix-signaling axis. This hypothesis was supported by significant changes in the gene expression profile of adhesion and extracellular matrix proteins in the Ptp4a3−/− CRC cells. The isogenic Ptp4a3fl/fl and Ptp4a3−/− CRC cells provide an invaluable tool to elucidate further the function of PTP4A3 and its link to adhesion and the extracellular matrix interactions. Citation Format: Kelley E. McQueeney, Paula Pekic, Joseph M. Salamoun, Jennifer Ahn, Elizabeth R. Sharlow, Peter Wipf, John S. Lazo. Depletion of PTP4A3 phosphatase disrupts colorectal cancer cell adhesion and extracellular matrix interactions. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 202.

Multiple roles of CD90 in cancer

THY1 (CD90) is a 25-37-kDa heavily N-glycosylated, glycophosphatidylinositol (GPI) anchored cell surface protein. It is usually expressed on thymocytes, mesenchymal stem cells, hematopoietic stem cells, natural killer cells, neurons, endothelial cells, renal glomerular mesangial cells, follicular dendritic cells, fibroblasts, and myofibroblasts. It has been found to regulate cell adhesion, migration, apoptosis, axon growth, cell-cell and cell-matrix interactions, T-cell activation, and fibrosis. Several reports have shown that CD90 has an important role in cancer in regulating cancer cell proliferation, metastasis, and angiogenesis. There are also evidences that CD90 is an important prognostic marker in many cancers. Consequently, therapies that target CD90 have great promise in treating many cancers. However, several studies also indicate a contradictory role for CD90, where it acts as a tumor suppressor. In this review, we summarize the expression, function of CD90 in different cancers and its possible use as a biomarker or a therapeutic target in cancer. The challenges and future prospects for the use of CD90 for clinical applications are also discussed in this review.

Structure-based site-directed photo-crosslinking analyses of multimeric cell-adhesive interactions of voltage-gated sodium channel β subunits.

The β1, β2, and β4 subunits of voltage-gated sodium channels reportedly function as cell adhesion molecules. The present crystallographic analysis of the β4 extracellular domain revealed an antiparallel arrangement of the β4 molecules in the crystal lattice. The interface between the two antiparallel β4 molecules is asymmetric, and results in a multimeric assembly. Structure-based mutagenesis and site-directed photo-crosslinking analyses of the β4-mediated cell-cell adhesion revealed that the interface between the antiparallel β4 molecules corresponds to that in the trans homophilic interaction for the multimeric assembly of β4 in cell-cell adhesion. This trans interaction mode is also employed in the β1-mediated cell-cell adhesion. Moreover, the β1 gene mutations associated with generalized epilepsy with febrile seizures plus (GEFS+) impaired the β1-mediated cell-cell adhesion, which should underlie the GEFS+ pathogenesis. Thus, the structural basis for the β-subunit-mediated cell-cell adhesion has been established.

Galectin-3 induces ovarian cancer cell survival and chemoresistance via TLR4 signaling activation

Paclitaxel resistance becomes common in patients with aggressive ovarian cancer and results in recurrence after conventional therapy. Galectin-3 is a multifunctional lectin associated with cell migration, cell proliferation, cell adhesion, and cell-cell interaction in tumor cells. Whether circulating galectin-3 is involved in paclitaxel resistance in ovarian cancer remains unknown. The current study investigated the effect of galectin-3 on toll-like receptor 4 (TLR4) signaling and thus paclitaxel resistance. With blood and cancer tissue samples obtained from 102 patients, we identified associations between serum galectin-3 level or TLR4 expression and paclitaxel resistance phenotype. In vitro, treatment with exogenous galectin-3 restored cell survival and migration of SKOV-3 and ES-2 cells was decreased by galectin-3 silencing and paclitaxel treatment. Furthermore, exogenous galectin-3 boosted expression of TLR4, MyD88, and p-p65, as well as interleukin (IL)-6, IL-8, and vascular endothelial growth factor (VEGF) release induced by paclitaxel. Moreover, galectin-3 inhibited the interaction between TLR4 and caveolin-1 (Cav-1) in SKOV-3 and ES-2 cells. In addition, overexpression of Cav-1 dampened the expression of MyD88 and p-p65 stimulated by galectin-3 and enhanced apoptosis in SKOV-3 cells under paclitaxel exposure. In summary, our study elucidated that exogenous galectin-3 might induce paclitaxel resistance through TLR4 signaling activation by inhibiting TLR4-Cav-1 interaction, revealing a novel insight into paclitaxel resistance induction.

Inclusion of gold nanoparticles in meso-porous silicon for the SERS analysis of cell adhesion on nano-structured surfaces

The study and the comprehension of the mechanism of cell adhesion and cell interaction with a substrate is a key point when biology and medicine meet engineering. This is the case of several biomedical applications, from regenerative medicine and tissue engineering to lab on chip and many others, in which the realization of the appropriate artificial surface allows the control of cell adhesion and proliferation.In this context, we aimed to design and develop a fabrication method of mesoporous (MeP) silicon substrates, doped with gold nanoparticles, in which we combine the capability of porous surfaces to support cell adhesion with the SERS capabilities of gold nanoparticles, to understand the chemical mechanisms of cell/surface interaction.MeP Si surfaces were realized by anodization of a Si wafer, creating the device for cell adhesion and growth. Gold nanoparticles were deposited on porous silicon by an electroless technique. We thus obtained devices with superior SERS capabilities, whereby cell activity may be controlled using Raman spectroscopy. MCF-7 breast cancer cells were cultured on the described substrates and SERS maps revealing the different expression and distribution of adhesion molecules were obtained by Raman spectroscopic analyses.

Polymer hydrogel particles as biocompatible AFM probes to study CD44/hyaluronic acid interactions on cells

Abstract We present a new AFM based methodology to detect specific interactions at cell surfaces. The central idea of this approach is to mimic ligand and receptor mediated physiological adhesion events at cell surfaces. Such adhesion events usually occur between mechanically compliant and hydrated surfaces. Consequently, we synthesized soft, ligand functionalized hydrogel particles and used them as colloidal AFM probes (soft colloidal probes, SCPs). As a proof-of-principle ligand/receptor system we studied the interaction between hyaluronic acid (HA) ligands and CD44 cell receptors. HA/CD44 interactions are involved in manifold signaling processes, in wound healing and tumor progression. Besides a detailed analysis of HA functionalized polyethylene glycol based SCPs, we show that these SCPs selectively bind to cells with CD44 receptors at their surface by means of AFM adhesion measurements. All negative controls showed strongly reduced adhesion forces demonstrating the feasibility of the method. Furthermore we analyzed the time dependence of CD44 and HA adhesion which indicated a rather fast development of the ligand/receptor interaction. Maximum adhesion was reached at about 10 s after contact providing further indication that HA/CD44 interactions can mediate early stages of cell adhesion. Overall this work shows that the specific adhesive interactions of cells can be analyzed close to the biological context by using hydrogel based colloidal AFM probes.

Comparative proteomic analysis by iTRAQ-2DLC-MS/MS provides insight into the key proteins involved in Cronobacter sp. biofilm formation

Cronobacter sp., an opportunistic foodborne pathogen, can cause high mortality in neonatal and premature infants because this pathogen is associated with powdered infant formula (PIF) contamination. Biofilm formation is recognized as the most effective means to help Cronobacter survive throughout the long shelf-life of PIF, thus promoting resistance to cleaning agents and disinfectants. Crystal violet (CV) staining and fluorescence microscopy analyses were used to compare the biofilm formation ability among different Cronobacter strains, and then the weak biofilm former Cronobacter sakazakii ATCC 29544 and the strong biofilm former Cronobacter dubliniensis DSM 18707 were selected for further proteomic analysis. Two-dimensional liquid chromatography–tandem mass spectrometry, which was coupled with isobaric tags for relative and absolute quantification (iTRAQ) labeling, was employed to quantitatively identify the proteins that were differentially expressed in C. sakazakii ATCC 29544 compared to C. dubliniensis DSM 18707. In total, 1190 proteins were detected. Gene Ontology (GO) analysis indicated that these differentially expressed proteins are related to biological binding, cell structure, signal transduction, cell adhesion, and cellular interaction. Among these differential proteins, the expression levels of 177 non-redundant proteins were altered significantly by more than 5-fold, with 96 up-regulated proteins and 81 down-regulated. This study provides important information regarding the cellular requirements for the underlying mechanism of Cronobacter biofilm formation and its survival in harsh environments.

Identification of genes associated with melanoma metastasis

The aims of the study were to identify the differentially expressed genes (DEGs) between primary melanomas and metastasis melanomas (MMs), and to investigate the mechanisms of MMs. The microarray data GSE8401 including 31 primary melanomas and 52 MMs were downloaded from Gene Expression Omnibus. DEGs were identified using the Linear Models for Microarray Data package. The functional and pathway enrichment analyses were performed for DEGs. Identification of transcription factors, tumor-associated genes (TAGs), and tumor suppressor genes (TSGs) were performed with the TRANSFAC, TAG, and TSGene databases, respectively. A protein–protein interaction network was constructed using Search Tool for the Retrieval of Interacting Genes. The modules construction and analysis was performed using Molecular Complex Detection and Gene Cluster with Literature Profiles, respectively. In total, 1004 upregulated and 1008 downregulated DEGs were identified. The upregulated DEGs, such as CDK1, BRCA1, MAD2L1, and PCNA, were significantly enriched in cell cycles, DNA replication, and mismatch repair. The downregulated DEGs, such as COLIAL, COL4A5, COL18A1, and LAMC2, were enriched in cell adhesion and extracellular matrix-receptor interaction. BRCA1 was identified as a transcription factor and TSG, and COL18A1 and LAMC2 were identified as a TSG and TAG, respectively. The upregulated DEGs had higher degrees in the protein–protein interaction network and module, such as PCNA, CDK1, and MAD2L1, and the heat map showed they were clustered in the functions of cell cycle and division. These results may demonstrate the potential roles of DEGs such as CDK1, BRCA1, COL18A1, and LAMC2 in the mechanism of MM.

Global Proteomic Profiling of the Secretome of Candida albicans ecm33 Cell Wall Mutant Reveals the Involvement of Ecm33 in Sap2 Secretion.

Candida albicans secretes numerous proteins related to cell wall remodeling, adhesion, nutrient acquisition and host interactions. Also, extracellular vesicles containing cytoplasmic proteins are secreted into the medium. The C. albicans ecm33/ecm33 mutant (RML2U) presents an altered cell wall and is avirulent. The proteomic analysis of proteins secreted by RML2U cells identified a total of 170 proteins: 114 and 154 of which correspond to the vesicle-free secretome and extracellular vesicles, respectively. Notably, 98 proteins were common to both samples, and the groups most represented were metabolic and cell wall-related proteins. The results of this study showed that RML2U had an altered pattern of proteins secreted by the classical secretion pathway as well as the formation of extracellular vesicles, including their size, quantity, and protein composition. Specifically, the secretion of aspartic protease 2 (Sap2) was compromised but not its intracellular expression, with bovine serum albumin (BSA) degradation by RML2U being altered when BSA was used as the sole nitrogen source. Furthermore, as recent research links the expression of Sap2 to the TOR (Target Of Rapamycin) signaling pathway, the sensitivity of RML2U to rapamycin (the inhibitor of TOR kinase) was tested and found to be enhanced, connecting Ecm33 with this pathway.

Rap1 GTPase is required for mouse lens epithelial maintenance and morphogenesis

Rap1, a Ras-like small GTPase, plays a crucial role in cell−matrix adhesive interactions, cell−cell junction formation, cell polarity and migration. The role of Rap1 in vertebrate organ development and tissue architecture, however, remains elusive. We addressed this question in a mouse lens model system using a conditional gene targeting approach. While individual germline deficiency of either Rap1a or Rap1b did not cause overt defects in mouse lens, conditional double deficiency (Rap1 cKO) prior to lens placode formation led to an ocular phenotype including microphthalmia and lens opacification in embryonic mice. The embryonic Rap1 cKO mouse lens exhibited striking defects including loss of E-cadherin- and ZO-1-based cell−cell junctions, disruption of paxillin and β1-integrin-based cell adhesive interactions along with abnormalities in cell shape and apical−basal polarity of epithelium. These epithelial changes were accompanied by increased levels of α-smooth muscle actin, vimentin and N-cadherin, and expression of transcriptional suppressors of E-cadherin (Snai1, Slug and Zeb2), and a mesenchymal metabolic protein (Dihydropyrimidine dehydrogenase). Additionally, while lens differentiation was not overtly affected, increased apoptosis and dysregulated cell cycle progression were noted in epithelium and fibers in Rap1 cKO mice. Collectively these observations uncover a requirement for Rap1 in maintenance of lens epithelial phenotype and morphogenesis.

PP09.10 – 2510: A novel case of periaxin-related neuropathy associated with cataract: Expanding the clinical and molecular spectrum of perxiaxin mutation

Objective Periaxin gene (PRX) is responsible for autosomal recessive early onset demyelinlating neuropathy: Charcot-Marie Tooth disease 4F (CMT4F) or its severe form Dejerine Sottas neuropathy (DSN). Periaxin protein is essential for maintenance of the peripheral nerve myelin probably in stabilizing its structure. It also functions as a scaffold protein that plays a role in lens membrane organization and cell adhesive interactions. Methods Exome sequencing. Results This report describes a child of non-consanguineous healthy parents with delayed motor milestones, frequent falls, and subcapsular cataract. His neurological exam revealed typical neuropathic features of the lower extremities: weakness of the lower limbs accompanied by absence of deep tendon reflexes. There was no sensory deficiency though he walked on abroad basis. Even so, his motor functions have improved over the years. Nerve conduction velocity (NCV) demonstrated demyelinating process with secondary axonal damage. Bilateral sub capsular cataract was discovered at one year old with no other ophthalmic pathology. A novel homozygote frame shift mutation c.418-419insG (P.m 149 fs) in the PRX gene was identified by whole exome sequencing and was confirmed by molecular analysis. Both his parents were found to be heterozygote carriers of this mutation. Conclusion Mutation in the periaxin protein is most probably the cause of peripheral distal neuropathy and cataract in this child. As of today, cataracts caused by mutations in the PRX gene were demonstrated only in laboratory animals. This is the first description of human congenital neuropathy coexistence with congenital cataract caused by mutations in the PRX gene. This is a new clinical presentation of PRX related hereditary neuropathy. It is suggested that careful eye examination should be done in every patients with congenital neuropathy. The coexistence of neuropathy and cataract may direct to the final diagnosis and its genetic cause.

Mechanokinetics of receptor–ligand interactions in cell adhesion

Receptor–ligand interactions in blood flow are crucial to initiate such biological processes as inflammatory cascade, platelet thrombosis, as well as tumor metastasis. To mediate cell adhesion, the interacting receptors and ligands must be anchored onto two apposing surfaces of two cells or a cell and a substratum, i.e., two-dimensional (2D) binding, which is different from the binding of a soluble ligand in fluid phase to a receptor, i.e., three-dimensional (3D) binding. While numerous works have been focused on 3D kinetics of receptor–ligand interactions in the immune system, 2D kinetics and its regulations have been less understood, since no theoretical framework or experimental assays were established until 1993. Not only does the molecular structure dominate 2D binding kinetics, but the shear force in blood flow also regulates cell adhesion mediated by interacting receptors and ligands. Here, we provide an overview of current progress in 2D binding and regulations, mainly from our group. Relevant issues of theoretical frameworks, experimental measurements, kinetic rates and binding affinities, and force regulations are discussed. A neutrophil undergoes capture and rolling (or tethering) on the endothelium through selectin–PSGL-1 bonds, followed by slow rolling and firm adhesion through the $${\upbeta }_{2}$$ -integrins LFA-1 and Mac-1 as well as intraluminal crawling and transmigration through the endothelium to the inflamed tissue.

High Foetal Haemoglobin in Sickle Cell Disease: Not so Protective?

High Foetal Haemoglobin in Sickle Cell Disease: Not so Protective?

글래스비드 정밀공급기구에 의한 바이오재료평가용 AFM프로브의 개발

Many different cells types have been found to be highly sensitive to mechanical force imposed by their surroundings. The cellular response to external mechanical forces has very important effects on numerous biological phenomena. In spite of its importance in biological processes, the cell adhesion force remains difficult to measure quantitatively at the cellular level. In this paper, to enhance quantitative measurements of cell adhesive interactions, a new attaching system and a method in which a glass bead can be attached to an AFM cantilever was designed and fabricated, and the degree of range displacement was controlled in the system. In an experiment, the movement of the stage in the attaching system and the attaching process were measured. The effectiveness of this system was confirmed as well in the experiment. In addition, through a commercial AFM system, the spring constant of the modified AFM probe could be measured.

Genetic and phenotypic intra-species variation in Candida albicans.

Candida albicans is a commensal fungus of the human gastrointestinal tract and a prevalent opportunistic pathogen. To examine diversity within this species, extensive genomic and phenotypic analyses were performed on 21 clinical C. albicans isolates. Genomic variation was evident in the form of polymorphisms, copy number variations, chromosomal inversions, subtelomeric hypervariation, loss of heterozygosity (LOH), and whole or partial chromosome aneuploidies. All 21 strains were diploid, although karyotypic changes were present in eight of the 21 isolates, with multiple strains being trisomic for Chromosome 4 or Chromosome 7. Aneuploid strains exhibited a general fitness defect relative to euploid strains when grown under replete conditions. All strains were also heterozygous, yet multiple, distinct LOH tracts were present in each isolate. Higher overall levels of genome heterozygosity correlated with faster growth rates, consistent with increased overall fitness. Genes with the highest rates of amino acid substitutions included many cell wall proteins, implicating fast evolving changes in cell adhesion and host interactions. One clinical isolate, P94015, presented several striking properties including a novel cellular phenotype, an inability to filament, drug resistance, and decreased virulence. Several of these properties were shown to be due to a homozygous nonsense mutation in the EFG1 gene. Furthermore, loss of EFG1 function resulted in increased fitness of P94015 in a commensal model of infection. Our analysis therefore reveals intra-species genetic and phenotypic differences in C. albicans and delineates a natural mutation that alters the balance between commensalism and pathogenicity.

Syndecans in angiogenesis and endothelial cell biology.

Syndecans are multifunctional heparan sulfate proteoglycans (HSPGs) with roles in cell adhesion, migration, receptor trafficking and growth-factor interactions and signalling. Studies using syndecan null animals have revealed limited roles for syndecans during development; however, under conditions of challenge or insult, several phenotypes have emerged. Angiogenesis is an important process both in development and in wound healing, but also in pathologies such as cancer and chronic inflammatory conditions. In the present paper, we summarize the main studies elucidating the role of syndecans in angiogenesis and their potential as novel therapeutic targets.

Ubiquinol decreases hemorrhagic shock/resuscitation-induced microvascular inflammation in rat mesenteric microcirculation.

Hemorrhagic shock (HS) is a leading cause of death in traumatic injury. Ischemia and hypoxia in HS and fluid resuscitation (FR) creates a condition that facilitates excessive generation of reactive oxygen species (ROS). This is a major factor causing increased leukocyte‐endothelial cell adhesive interactions and inflammation in the microcirculation resulting in reperfusion tissue injury. The aim of this study was to determine if ubiquinol (coenzyme Q10) decreases microvascular inflammation following HS and FR. Intravital microscopy was used to measure leukocyte‐endothelial cell adhesive interactions in the rat mesentery following 1‐h of HS and 2‐h post FR with or without ubiquinol. Hemorrhagic shock was induced by removing ~ 40% of anesthetized Sprague Dawley rats' blood volume to maintain a mean arterial blood pressure <50 mmHg for 1 h. Ubiquinol (1 mg/100 g body weight) was infused intravascularly in the ubiquinol group immediately after 1‐h HS. The FR protocol included replacement of the shed blood and Lactate Ringer's in both the control and ubiquinol groups. We found that leukocyte adherence (2.3 ± 2.0), mast cell degranulation (1.02 ± 0.01), and ROS levels (159 ± 35%) in the ubiquinol group were significantly reduced compared to the control group (10.8 ± 2.3, 1.36 ± 0.03, and 343 ± 47%, respectively). In addition, vascular permeability in the control group (0.54 ± 0.11) was significantly greater than the ubiquinol group (0.34 ± 0.04). In conclusion, ubiquinol attenuates HS and FR‐induced microvascular inflammation. These results suggest that ubiquinol provides protection to mesenteric microcirculation through its antioxidant properties.

Computational modelling of epidermal stratification highlights the importance of asymmetric cell division for predictable and robust layer formation.

Skin is a complex organ tasked with, among other functions, protecting the body from the outside world. Its outermost protective layer, the epidermis, is comprised of multiple cell layers that are derived from a single-layered ectoderm during development. Using a new stochastic, multi-scale computational modelling framework, the anisotropic subcellular element method, we investigate the role of cell morphology and biophysical cell-cell interactions in the formation of this layered structure. This three-dimensional framework describes interactions between collections of hundreds to thousands of cells and (i) accounts for intracellular structure and morphology, (ii) easily incorporates complex cell-cell interactions and (iii) can be efficiently implemented on parallel architectures. We use this approach to construct a model of the developing epidermis that accounts for the internal polarity of ectodermal cells and their columnar morphology. Using this model, we show that cell detachment, which has been previously suggested to have a role in this process, leads to unpredictable, randomized stratification and that this cannot be abrogated by adjustment of cell-cell adhesion interaction strength. Polarized distribution of cell adhesion proteins, motivated by epithelial polarization, can however eliminate this detachment, and in conjunction with asymmetric cell division lead to robust and predictable development.

Nicotinamide phosphoribosyltransferase can affect metastatic activity and cell adhesive functions by regulating integrins in breast cancer.

NAD(+) metabolism is an essential regulator of cellular redox reactions, energy pathways, and a substrate provider for NAD(+) consuming enzymes. We recently demonstrated that enhancement of NAD(+)/NADH levels in breast cancer cells with impaired mitochondrial NADH dehydrogenase activity, through augmentation of complex I or by supplementing tumor cell nutrients with NAD(+) precursors, inhibits tumorigenicity and metastasis. To more fully understand how aberrantly low NAD(+) levels promote tumor cell dissemination, we here asked whether inhibition of NAD(+) salvage pathway activity by reduction in nicotinamide phosphoribosyltransferase (NAMPT) expression can impact metastasis and tumor cell adhesive functions. We show that knockdown of NAMPT, the enzyme catalyzing the rate-limiting step of the NAD(+) salvage pathway, enhances metastatic aggressiveness in human breast cancer cells and involves modulation of integrin expression and function. Reduction in NAMPT expression is associated with upregulation of select adhesion receptors, particularly αvβ3 and β1 integrins, and results in increased breast cancer cell attachment to extracellular matrix proteins, a key function in tumor cell dissemination. Interestingly, NAMPT downregulation prompts expression of integrin αvβ3 in a high affinity conformation, known to promote tumor cell adhesive interactions during hematogenous metastasis. NAMPT has been selected as a therapeutic target for cancer therapy based on the essential functions of this enzyme in NAD(+) metabolism, cellular redox, DNA repair and energy pathways. Notably, our results indicate that incomplete inhibition of NAMPT, which impedes NAD(+) metabolism but does not kill a tumor cell can alter its phenotype to be more aggressive and metastatic. This phenomenon could promote cancer recurrence, even if NAMPT inhibition initially reduces tumor growth.

Anabolic androgens affect the competitive interactions in cell migration and adhesion between normal mouse urothelial cells and urothelial carcinoma cells

The urothelium is constantly rebuilt by normal urothelial cells to regenerate damaged tissues caused by stimuli in urine. However, the urothelial carcinoma cells expand the territory by aberrant growth of tumor cells, which migrate and occupy the damaged tissues to spread outside and disrupt the normal cells and organized tissues and form a tumor. Therefore, the interaction between normal urothelial cells and urothelial carcinoma cells affect the initiation and progression of urothelial tumors if normal urothelial cells fail to migrate and adhere to the damages sites to regenerate the tissues. Here, comparing normal murine urothelial cells with murine urothelial carcinoma cells (MBT-2), we found that normal cells had less migration ability than carcinoma cells. And in our co-culture system we found that carcinoma cells had propensity migrating toward normal urothelial cells and carcinoma cells had more advantages to adhere than normal cells. To reverse this condition, we used anabolic androgen, dihyrotestosterone (DHT) to treat normal cells and found that DHT treatment increased the migration ability of normal urothelial cells toward carcinoma cells and the adhesion capacity in competition with carcinoma cells. This study provides the base of a novel therapeutic approach by using anabolic hormone-enforced normal urothelial cells to regenerate the damage urothelium and defend against the occupancy of carcinoma cells to thwart cancer development and recurrence.