The cancer cell metastasis is a major death reason for patients with non-small cell lung cancer(NSCLC).Although researchers have disclosed that interleukin 17(IL-17)can increase matrix metalloproteinases(MMPs)inductio...The cancer cell metastasis is a major death reason for patients with non-small cell lung cancer(NSCLC).Although researchers have disclosed that interleukin 17(IL-17)can increase matrix metalloproteinases(MMPs)induction causing NSCLC cell metastasis,the underlying mechanism remains unclear.In the study,we found that IL-17 receptor A(IL-17RA),p300,p-STAT3,Ack-STAT3,and MMP19 were up-regulated both in NSCLC tissues and NSCLC cells stimulated with IL-17.p300,STAT3 and MMP19 overexpression or knockdown could raise or reduce IL-17-induced p-STAT3,Ack-STAT3 and MMP19 level as well as the cell migration and invasion.Mechanism investigation revealed that STAT3 and p300 bound to the same region(−544 to−389 nt)of MMP19 promoter,and p300 could acetylate STAT3-K631 elevating STAT3 transcriptional activity,p-STAT3 or MMP19 expression and the cell mobility exposed to IL-17.Meanwhile,p300-mediated STAT3-K631 acetylation and its Y705-phosphorylation could interact,synergistically facilitating MMP19 gene transcription and enhancing cell migration and invasion.Besides,the animal experiments exhibited that the nude mice inoculated with NSCLC cells by silencing p300,STAT3 or MMP19 gene plus IL-17 treatment,the nodule number,and MMP19,Ack-STAT3,or p-STAT3 production in the lung metastatic nodules were all alleviated.Collectively,these outcomes uncover that IL-17-triggered NSCLC metastasis involves up-regulating MMP19 expression via the interaction of STAT3-K631 acetylation by p300 and its Y705-phosphorylation,which provides a new mechanistic insight and potential strategy for NSCLC metastasis and therapy.展开更多
Runx2 is a major regulator of osteoblast differentiation and function;however,the role of Runx2 in peripheral nerve repair is unclea r.Here,we analyzed Runx2expression following injury and found that it was specifical...Runx2 is a major regulator of osteoblast differentiation and function;however,the role of Runx2 in peripheral nerve repair is unclea r.Here,we analyzed Runx2expression following injury and found that it was specifically up-regulated in Schwann cells.Furthermore,using Schwann cell-specific Runx2 knocko ut mice,we studied peripheral nerve development and regeneration and found that multiple steps in the regeneration process following sciatic nerve injury were Runx2-dependent.Changes observed in Runx2 knoc kout mice include increased prolife ration of Schwann cells,impaired Schwann cell migration and axonal regrowth,reduced re-myelination of axo ns,and a block in macrophage clearance in the late stage of regeneration.Taken together,our findings indicate that Runx2 is a key regulator of Schwann cell plasticity,and therefore peripheral nerve repair.Thus,our study shows that Runx2 plays a major role in Schwann cell migration,re-myelination,and peripheral nerve functional recovery following injury.展开更多
Collective cancer cell migration(CCCM)and epithelial-to-mesenchymal transition(EMT)play key roles in metastasis.This study reports that the colorectal carcinoma cell line LIM1863 is useful for the study of CCCM and EM...Collective cancer cell migration(CCCM)and epithelial-to-mesenchymal transition(EMT)play key roles in metastasis.This study reports that the colorectal carcinoma cell line LIM1863 is useful for the study of CCCM and EMT.Methods:Hematoxylin and eosin staining,scanning electron microscopy,transmission electron microscopy,and western blot analysis were performed.Results:LIM1863 automatically grew as spheroids in suspension and had important typical epithelial properties,including several layers of cells arranged around a central lumen,apical-basal polarity,and types of cell-cell junctions.Treatment with a combination of both TGF beta 1 and TNF alpha induced definite and distinct EMT,a spheroid changing phenotype to form a monolayer high-confluent patch without lumen,without polarity.Spontaneous CCCM occurred in spheroids.Flat EMT cells adhered to the base of a dish,exhibited persistent movement as a cluster of cells,and then shed,resulting in a cluster.All cells from one cluster undergoing CCCM died.Otherwise,all cells undergoing EMT disappeared and almost all cells located in the cell reservoir survived and proliferated.Conclusion:LIM1863 is an excellent cell line to study CCCM and EMT.The group of heterogeneous cells undergoing CCCM behaves like a supracellular unit.展开更多
As a pathway that plays a role in nutrient absorption,anabolic response,cell growth and survival,the important role of AKT/mTOR in tumorigenesis has also come to light.For cancer patients,most deaths are caused by the...As a pathway that plays a role in nutrient absorption,anabolic response,cell growth and survival,the important role of AKT/mTOR in tumorigenesis has also come to light.For cancer patients,most deaths are caused by the growth of metastatic tumors outside the primary focus.Therefore,migration and invasion in the late stage of tumor progression are the main unresolved issues in the study of tumor pathogenesis,and AKT/mTOR has been found to participate in the migration and invasion of cancer cells,which means that the study of this pathway may contribute to a solution for the problem.Because of its extensive and complex functions in the organism,this pathway can be regulated by a variety of different signals in the body,and then realize its function through different downstream signal molecules.This article reviews the proteins that can indirectly affect this pathway by regulating the common upstream signaling molecules of this pathway,and the proteins that can directly affect the level of phosphorylation of AKT/mTOR in cancer cells.We also review the proteins that can co-regulate this pathway and its downstream pathways.Through this study,we hope to gain a deeper understanding of the regulatory mechanism of the AKT/mTOR pathway in cancer cells,in hopes of finding effective and harmless cancer treatment targets in the future.展开更多
Circular RNAs(circRNAs)play a vital role in diabetic peripheral neuropathy.However,their expression and function in Schwann cells in individuals with diabetic peripheral neuropathy remain poorly understood.Here,we per...Circular RNAs(circRNAs)play a vital role in diabetic peripheral neuropathy.However,their expression and function in Schwann cells in individuals with diabetic peripheral neuropathy remain poorly understood.Here,we performed protein profiling and circRNA sequencing of sural nerves in patients with diabetic peripheral neuropathy and controls.Protein profiling revealed 265 differentially expressed proteins in the diabetic peripheral neuropathy group.Gene Ontology indicated that differentially expressed proteins were mainly enriched in myelination and mitochondrial oxidative phosphorylation.A real-time polymerase chain reaction assay performed to validate the circRNA sequencing results yielded 11 differentially expressed circRNAs.circ_0002538 was markedly downregulated in patients with diabetic peripheral neuropathy.Further in vitro experiments showed that overexpression of circ_0002538 promoted the migration of Schwann cells by upregulating plasmolipin(PLLP)expression.Moreover,overexpression of circ_0002538 in the sciatic nerve in a streptozotocin-induced mouse model of diabetic peripheral neuropathy alleviated demyelination and improved sciatic nerve function.The results of a mechanistic experiment showed that circ_0002538 promotes PLLP expression by sponging miR-138-5p,while a lack of circ_0002538 led to a PLLP deficiency that further suppressed Schwann cell migration.These findings suggest that the circ_0002538/miR-138-5p/PLLP axis can promote the migration of Schwann cells in diabetic peripheral neuropathy patients,improving myelin sheath structure and nerve function.Thus,this axis is a potential target for therapeutic treatment of diabetic peripheral neuropathy.展开更多
Interconnectivity is the key characteristic of bone tissue engineering scaffold modulating cell migration,blood vessels invasion and transport of nutrient and waste.However,efforts and understanding of the interconnec...Interconnectivity is the key characteristic of bone tissue engineering scaffold modulating cell migration,blood vessels invasion and transport of nutrient and waste.However,efforts and understanding of the interconnectivity of porous Mg is limited due to the diverse architectures of pore struts and pore size distribution of Mg scaffold systems.In this work,biomimetic hierarchical porous Mg scaffolds with tailored interconnectivity as well as pore size distribution were prepared by template replication of infiltration casting.Mg scaffold with better interconnectivity showed lower mechanical strength.Enlarging interconnected pores would enhance the interconnectivity of the whole scaffold and reduce the change of ion concentration,pH value and osmolality of the degradation microenvironment due to the lower specific surface area.Nevertheless,the degradation rates of five tested Mg scaffolds were no different because of the same geometry of strut unit.Direct cell culture and evaluation of cell density at both sides of four typical Mg scaffolds indicated that cell migration through hierarchical porous Mg scaffolds could be enhanced by not only bigger interconnected pore size but also larger main pore size.In summary,design of interconnectivity in terms of pore size distribution could regulate mechanical strength,microenvironment in cell culture condition and cell migration potential,and beyond that it shows great potential for personalized therapy which could facilitate the regeneration process.展开更多
Cell migration is facilitated by the interaction of living cells and their local microenvironment.The local topography is one of the key factors regulating cell migration.Interaction between the surface topography and...Cell migration is facilitated by the interaction of living cells and their local microenvironment.The local topography is one of the key factors regulating cell migration.Interaction between the surface topography and the cell behaviors is critical to understanding tissue development and regeneration.In this study,a dynamic mask photolithography technique has been utilized to fabricate a surface with graded micropillars.It has been demonstrated that the cells have been successfully guided to migrate from the sparse zone to the dense zone.The cell polarization angle has been characterized in both sparse zone and the dense zone.Compared to the dense zone,the cells in the sparse zone are more aligned along the direction of the micropillar spacing gradient,which enables the guided cell migration.Moreover,the effects of the micropillar spacing gradient,micropillar diameter,and micropillar height have been investigated in terms of the cell migration speed and cell spreading area.Finally,two issues significantly affecting the cell migration have been discussed:trapped cells between the micropillars and cell clusters.展开更多
Cell migration in anisotropic microenvironment plays an important role in the development of normal tissues and organs as well as neoplasm progression,e.g.,osteogenic differentiation of embryonic stem cells was facili...Cell migration in anisotropic microenvironment plays an important role in the development of normal tissues and organs as well as neoplasm progression,e.g.,osteogenic differentiation of embryonic stem cells was facilitated on stiffer substrates,indicating that the mechanical signals greatly affect both early and terminal differentiation of embryonic stem cells.However,the effect of anisotropy on cell migration dynamics,in particular,in terms of acceleration profiles which is important for recognizing dynamics modes of cell migration and analyzing the regulation mechanisms of microenvironment in mechanical signal transmission,has not been systematically investigated.In this work,we firstly rigorously investigate and quantify the differences between persistent random walk and anisotropic persistent random walk models based on the analysis of cell migration trajectories and velocity auto-covariance function,both qualitatively and quantitatively.Secondly,we introduce the concepts of positive and negative anisotropy based on the motility parameters to study the effect of anisotropy on acceleration profiles,especially the nonlinear decrease and non-monotonic behaviors.We particularly elaborate and discuss the mechanisms,and physical insights of non-monotonic behaviors in the case of positive anisotropy,focusing on the force exerted on migrating cells.Finally,we analyze two types of in vitro cell migration experiments and verify the universality of nonlinear decrease and the consistence of non-monotonic behaviors with numerical results.We conclude that the anisotropy of microenvironment is the cause of the non-monotonic and nonlinear dynamics,and the anisotropic persistent random walk can be as a suitable tool to analyze in vitro cell migration with different combinations of motility parameters.Our analysis provides new insights into the dynamics of cell migration in complex microenvironment,which also has implications in tissue engineering and cancer research.展开更多
Cell migration plays a significant role in many biological activities,yet the physical mechanisms of cell migration are still not well understood.In this study,a continuum physics-based epithelial monolayer model incl...Cell migration plays a significant role in many biological activities,yet the physical mechanisms of cell migration are still not well understood.In this study,a continuum physics-based epithelial monolayer model including the intercellular interaction was employed to study the cell migration behavior in a confluent epithelial monolayer at constant cell density.The epithelial cell was modeled as isotropic elastic material.Through finite element simulation,the results revealed that themotile cellwas subjected to higher stress than the other jammed cells during the migration process.Cell stiffness was implied to play a significant role in epithelial cell migration behavior.Higher stiffness results in smaller displacement and lower migration speed.展开更多
The collective cell migration behavior on a substrate was studied using RKPM meshfree method.The cells were modeled as nematic liquid crystal with hyperelastic cell nucleus.The cell-substrate and cell-cell interaction...The collective cell migration behavior on a substrate was studied using RKPM meshfree method.The cells were modeled as nematic liquid crystal with hyperelastic cell nucleus.The cell-substrate and cell-cell interactions were modeled by coarse-grained potential forces.Through this study,the pulling and pushing phenomenon during collective cell migration process was observed and it was found that the individual cell mobility significantly influenced the collective cell migratory behavior.More self-propelled cells are in the system along the same direction,the faster the collective group migrates toward coordinated direction.The parametric study on cell-cell adhesion strength indicated that as the adhesion strength increases,the collective cell migration speed increases.It also showed that the mechanical stress in leader cell is higher than stress in follower cells.展开更多
The link of the metazoan nucleus to the actin cytoskeleton is highly important for actin polymerization and migration of multiple cell types as well as for mechanotransduction and even affects the cellular transcripto...The link of the metazoan nucleus to the actin cytoskeleton is highly important for actin polymerization and migration of multiple cell types as well as for mechanotransduction and even affects the cellular transcriptome.Several mechanisms of organization of actin filaments next to the nuclear envelope have been identified.Among these mechanisms the most studied one is the Linker of nucleoskeleton and cytoskeleton(LINC)complex-dependent perinuclear actin organization.However,recently additional mechanisms have been identified:an Actin-related protein-2/3(Arp2/3)-dependent perinuclear actin polymerization during migration of dendritic cells and a perinuclear actin rim that is formed in response to external force application or migration cues.In parallel,there are also reports on cancer cells that migrate in a LINC complex independent manner and on cancers with reduced expression of the LINC complex components.Thus,suggesting that LINC complex independent migration may be associated with tumour formation.展开更多
Abstract Meniscus injury is a common disease in clinic.If it was not treated in time,it leads to osteoarthritis which brings unbearable pain and heavy economic burden to the patients.At present,meniscectomy and menisc...Abstract Meniscus injury is a common disease in clinic.If it was not treated in time,it leads to osteoarthritis which brings unbearable pain and heavy economic burden to the patients.At present,meniscectomy and meniscus suture are widely used in the treatment for meniscus injury.Nevertheless,It is not ideal for poor self-healing ability of meniscus.The recruitment of endogenous stem cells is an attractive option for wounded meniscus healing.Fully reduced high-mobility group box 1 protein(HMGB1)can accelerate the regeneration of multiple tissues by endogenous stem cell activation,migration and differentiation.Kartogenin(KGN)has shown to induce the chondrogenesis of the stem cells.However,no study has explored such effects of HMGB1 and KGN in wounded meniscus healing.Therefore,in order to improve the regeneration of meniscus,we intend to use a novel bioactive microsphere which was developed by combining fully reduced high mobility group box 1(frHMGB1)and kartogenin(KGN)with alginate gel which slowly release high concentrations of HMGB1 and KGN to activate rat bone marrow stem cells(BMSCs)and promote cell proliferation.The results showed that this HMGB1–KGN microsphere released and kept high concentrations of HMGB1 and KGN in the wound area for more than 2 weeks.In vitro experimental results showed that the HMGB1–KGN microsphere can promote cell proliferation via recruiting rat bone marrow stem cells(BMSCs)and activating the BMSCs from G_(0) to G_(Alert) stage as evidenced by cell migration testing and 5-bromo-2′-deoxyuridine(BrdU)incorporation assay.In vivo results indicated that this HMGB-KGN microsphere can recruit GFP-labeled BMSCs from tail vein to wounded meniscus and induce these GFP-labeled BMSCs to differentiate into chondrocytes.Our results demonstrated that the HMGB1–KGN-containing bioactive microsphere induced cell migration in vitro and recruited the cells to wound area to promote wounded rat meniscus healing in vivo.展开更多
Distal gastric cancer(DGC)is a subgroup of gastric cancer(GC),which has different molecular characteristics from proximal gastric cancer(PGC).These differences result in different overall survival(OS)rates;however,dat...Distal gastric cancer(DGC)is a subgroup of gastric cancer(GC),which has different molecular characteristics from proximal gastric cancer(PGC).These differences result in different overall survival(OS)rates;however,data pertaining to the survival rate in PGC or DGC are contradictory.This suggests that the location of GC is not the unique cause of the different survival rates,while the molecular characteristics might be more important factors determining the prognosis of DGC.Therefore,the aim of this study was to discover key prognostic factors in DGC using bioinformatic methods and to explore the potential molecular mechanism.The Cancer Genome Atlas(TCGA)public database was employed to screen data relating to DGC,and we conducted a weighted gene co-expression network analysis(WGCNA)on DGC patient samples to establish co-expression modules.High-weight genes(hub genes)in a dominant color module were identified.In vitro experiments and gene set enrichment analyses(GSEA)were carried out to elucidate the potential molecular mechanism.In this study,139 DGC samples were enrolled to perform a co-expression analysis.According to the correlation between gene modules and clinical characteristics,the royal blue module related to stage M of DGC was screened,and a survival analysis was conducted to show that highcoagulation-factor V(F5)expression was related to the short OS of patients with GC.In vitro experiments confirmed that F5 could promote the migration of GC cells.GSEA suggested that F5 might have affected the prognosis of GC by modulating the activities of the Wnt and/or the TGF-βsignaling pathways.Our results indicated that high F5 expression predicts poor prognosis of patients with DGC,and it functions probably by promoting cell migration through the Wnt and/or the TGF-βsignaling pathways.展开更多
Much of our understanding of the events which underlie cell migration has been derived from studies of cells intissue culture. One of the components that mediates this process is the dynamic actin-based microfilament ...Much of our understanding of the events which underlie cell migration has been derived from studies of cells intissue culture. One of the components that mediates this process is the dynamic actin-based microfilament system that canreorganize itself into so-called stress fibers that are considered essential components for cell motility. In contrast, relativelyfew studies have investigated cell movement along an extracellular matrix (ECM) which is known to influence both cellularorganization and behavior. This opinion/viewpoint article briefly reviews cell migration during corneal endothelial woundrepair along the tissue’s natural basement membrane, Descemet’s membrane. Because the tissue exists as a cell monolayer itaffords one an opportunity to readily explore the effect of cell/matrix influences on cell motility. As such, cell movementalong this substrate differs somewhat from that found in vitro and migrating endothelial cells also demonstrate an abilityto move along the ECM without the benefit of having an organized actin cytoskeleton.展开更多
Migration of dendritic cells (DCs) into tissues and secondary lymphoid organs plays a crucial role in the initiation of innate and adaptive immunity. In this article, we show that cyclosporin A (CsA) impairs the migra...Migration of dendritic cells (DCs) into tissues and secondary lymphoid organs plays a crucial role in the initiation of innate and adaptive immunity. In this article, we show that cyclosporin A (CsA) impairs the migration of DCs both in vitro and in vivo. Exposure of DCs to clinical concentrations of CsA neither induces apoptosis nor alters development but does impair cytokine secretion, chemokine receptor expression, and migration. In vitro, CsA impairs the migration of mouse bone marrow-derived DCs toward macrophage inflammatory protein-3beta (MIP-3beta) and induces them to retain responsiveness to MIP-1alpha after lipopolysaccharide (LPS)-stimulated DC maturation, while in vivo administration of CsA inhibits the migration of DCs out of skin and into the secondary lymphoid organs. CsA impairs chemokine receptor and cyclooxygenase-2 (COX-2) expression normally triggered in LPS-stimulated DCs; administration of exogenous prostaglandin E2 (PGE2) reverses the effects of CsA on chemokine receptor expression and DC migration. Inhibition of nuclear factor-kappaB (NF-κB) and mitogen-activated protein kinase (MAPK) pathway signaling by CsA may be responsible for the CsA-mediated effects on the regulation of chemokine receptor and cyclooxygenase-2 (COX-2) expression. Impairment of DC migration due to inhibition of PGE2 production and regulation of chemokine receptor expression may contribute, in part, to CsA-mediated immunosuppression.展开更多
Background To investigate the effects and mechanisms of cellular repressor of ElA stimulated genes (CREG) on endothelial cell(EC) migration.Methods vascular endothelial cells(VE),CREG overexpression VEs, CREG suppress...Background To investigate the effects and mechanisms of cellular repressor of ElA stimulated genes (CREG) on endothelial cell(EC) migration.Methods vascular endothelial cells(VE),CREG overexpression VEs, CREG suppression VEs and VEs transfected with CREG gene modified adenovirus(Ad-CREG) were cultured with dulbecco’s modified eagle’s medium contained 10%fetal calf serum. Western blot was used to detect the protein level of CREG and integrin-linked kinase(ILK) in the four kind ECs.Tran-swell migration model was applied to compare the migration cell number of the four kind ECs.Two kinds of ILK mutant plasmids;PCXN2-flag-ILK wt-IRES-GFP(wild-type ILK)and PCXN2-flag-ILK p-parvin-IRES-GFP(P-parvin-binding mutant) were used to transfect VS and VE respectively,then the two kind transfection ECs were named as VS-wtILK and VE-P -parvin which were selected by G418(600ng/ml)for 2 weeks;Transwell migration model was applied to compare migration capability before and after ILK plasmids transfecting VE and VS.Results Western blot analysis showed that CREG overexpression promoted ILK expression in ECs,on the contrary,ILK expression was down-regulated in CREG silent ECs(P【0.05).Further more,ILK expression was up-regulated obviously in VE transfected with Ad-CREG(P【 0.05);Transwell migration model showed that EC’s migration capability was positively correlated with the expression level of CREG in EC,that is,CREG overexpression induced VE migration and CREG silent suppressed VE migration, moreover,Ad-CREG transfecting VE showed better migration capability accompanied with CREG expression increase by transwell migration model(P【0.05).In order to know the relationship between ILK expression and cell migration,we obtained stable transfection cell strains of VS-wtILK and VE-Pparvin, transwell migration model demonstrated that VS-wtILK remarkably corrected the poor migration capability of VS(P【 0.01),butβ-parvin combining site mutation in ILK genes inhibited VE migration markedly(P【0.01).Conclusions ILKp -parvin signal pathway mediated vascular endothelial cell migration induced by CREG.展开更多
AIM:To investigate the role of claudin 1 in the regulation of genes involved in cell migration and tumor necrosis factor alpha(TNF-α)-induced gene expression in human gastric adenocarcinoma cells.METHODS:Knockdown ex...AIM:To investigate the role of claudin 1 in the regulation of genes involved in cell migration and tumor necrosis factor alpha(TNF-α)-induced gene expression in human gastric adenocarcinoma cells.METHODS:Knockdown experiments were conducted with claudin 1 small interfering RNA(si RNA),and theeffects on the cell cycle,apoptosis,migration and invasion were analyzed in human gastric adenocarcinoma MKN28 cells.The gene expression profiles of cells were analyzed by microarray and bioinformatics.RESULTS:The knockdown of claudin 1 significantly inhibited cell proliferation,migration and invasion,and increased apoptosis.Microarray analysis identified 245genes whose expression levels were altered by the knockdown of claudin 1.Pathway analysis showed that the top-ranked molecular and cellular function was the cellular movement related pathway,which involved MMP7,TNF-SF10,TGFBR1,and CCL2.Furthermore,TNFand nuclear frctor-κB were the top-ranked upstream regulators related to claudin 1.TNF-αtreatment increased claudin 1 expression and cell migration in MKN28 cells.Microarray analysis indicated that the depletion of claudin1 inhibited 80%of the TNF-α-induced m RNA expression changes.Further,TNF-αdid not enhance cell migration in the claudin 1 si RNA transfected cells.CONCLUSION:These results suggest that claudin 1 is an important messenger that regulates TNF-α-induced gene expression and migration in gastric cancer cells.A deeper understanding of these cellular processes may be helpful in establishing new therapeutic strategies for gastric cancer.展开更多
Objective To investigate the effects of microRNA-18a(miR-18a) on migration and invasion of hepatocellular carcinoma(HCC) cells,and its possible mechanism associated with Dicer l. Methods HepG 2 and HepG 2.2.15 cells w...Objective To investigate the effects of microRNA-18a(miR-18a) on migration and invasion of hepatocellular carcinoma(HCC) cells,and its possible mechanism associated with Dicer l. Methods HepG 2 and HepG 2.2.15 cells were transfected with miR-18 a inhibitor using Lipofectamine. Cell invasion was evaluated by transwell invasion assay,and cell migration was detected by transwell migration and wound-healing assays. Moreover,luciferase reporter assay was used to identify whether Dicer expression was regulated by miR-18 a. Real-time RT-PCR and western blot were performed to analyze Dicer 1 expression. In addition,a functional restoration assay was performed to investigate whether miR-18 a promotes HCC cell migration and invasion by directly targeting Dicer 1. Results miR-18 a inhibitor can suppress the migration and invasion of HCC cells. Furthermore,suppression of Dicer l expression by small interfering RNA essentially abolished the inhibition of cell migration and invasion induced by miR-18 a inhibitor,restorating these activities to levels similar to the parental HCC cells. Interestingly,suppression of miR-18 a in HCC cells resulted in enhanced expression of Dicer l. In addition,the results of a luciferase assay demonstrated targeted regulation of Dicer l by miR-18 a. Conclusion Our findings suggest that miR-18 a promotes migration and invasion of HCC cells by inhibiting Dicer l expression.展开更多
The migration mode transition of cancer cell enhances its invasive capability and the drug resistance,where physical confinement of cell microenvironment has been revealed to induce the mesenchymal-amoeboid transition...The migration mode transition of cancer cell enhances its invasive capability and the drug resistance,where physical confinement of cell microenvironment has been revealed to induce the mesenchymal-amoeboid transition(MAT).However,most existing studies are performed in PDMS microchannels,of which the stiffness is much higher than that of most mammalian tissues.Therefore,the amoeboid migration transition observed in these studies is actually induced by the synergistic effect of matrix stiffness and confinement.Since the stiffness of cell microenvironment has been reported to influence the cell migration in 2D substrate,the decoupling of stiffness and confinement effects is thus in need for elucidating the underlying mechanism of MAT.However,it is technically challenging to construct microchannels with physiologically relevant stiffness and channel size,where existing microchannel platforms with physiological relevance stiffness are all with>10μm channel width.Such size is too wide to mimic the physical confinement that migrating cancer cells confront in vivo,and also larger than the width of PDMS channel,in which the MAT of cancer cell was observed.Therefore,an in vitro cell migration platform,which could mimic both stiffness and confinement of the native physical microenvironment during cancer metastasis,could profoundly contribute to researches on cancer cell migration and cellular mechanotransduction.In this paper,we overcome the limitations of engineering soft materials in microscale by combining the collagen-alginate hydrogel with photolithography.This enables us to improve the accuracy of molded microchannel,and thus successfully construct a 3D microchannel platform,which matches the stiffness and width ranges of native environmental confinement that migrating cancer cells confront in vivo.The stiffness(0.3~20 kPa),confinement(channel width:3.5~14μm)and the adhesion ligand density of the microchannel can be tuned independently.Interestingly,using this platform,we observed that the migration speed of cancer cell is influenced by the synergistic effect of channel stiffness and width,and the increasing stiffness reverses the effect of channel width on the migration speed of cancer cells.In addition,MAT has a strong correlation with the channel stiffness.These findings make us reconsider the widely accepted hypothesis:physical confinement can induce MAT.Actually,this transition can only occur in stiff confined microenvironment not in soft one.For soft microchannels,the compliance of the channel walls could cause little cell/nucleus deformation,and the MAT could not be induced.To further investigate the mechanism of MAT,we developed a computational model to simulate the effect of nucleus deformation on MAT.With the model,we found that deforming the cell nuclear by decreasing the nucleus stiffness will reduce the cellmigration speed.This implies that nuclear stiffness plays an important role in the regulation of cancer migration speed and thus MAT in microchannels.The effect of channel stiffness on MAT and migration speed as observed in our experiment could partially explain previous findings reported in the literature,where the increasing matrix stiffness of tumor microenvironment promotes cancer metastasis.Our observations thus highlight the critical role of cell nuclear deformation not only in MAT,but also in regulating cellular mechanotransduction and cell-ECM interactions.This developed platform is capable of mimicking the native physical microenvironment during metastasis,providing a powerful tool for high-throughput screening applications and investigating the interaction between cancer migration and biophysical microenvironment.展开更多
基金National Natural Science Foundation of China(Grants Numbers 81902878 and 81971468).
文摘The cancer cell metastasis is a major death reason for patients with non-small cell lung cancer(NSCLC).Although researchers have disclosed that interleukin 17(IL-17)can increase matrix metalloproteinases(MMPs)induction causing NSCLC cell metastasis,the underlying mechanism remains unclear.In the study,we found that IL-17 receptor A(IL-17RA),p300,p-STAT3,Ack-STAT3,and MMP19 were up-regulated both in NSCLC tissues and NSCLC cells stimulated with IL-17.p300,STAT3 and MMP19 overexpression or knockdown could raise or reduce IL-17-induced p-STAT3,Ack-STAT3 and MMP19 level as well as the cell migration and invasion.Mechanism investigation revealed that STAT3 and p300 bound to the same region(−544 to−389 nt)of MMP19 promoter,and p300 could acetylate STAT3-K631 elevating STAT3 transcriptional activity,p-STAT3 or MMP19 expression and the cell mobility exposed to IL-17.Meanwhile,p300-mediated STAT3-K631 acetylation and its Y705-phosphorylation could interact,synergistically facilitating MMP19 gene transcription and enhancing cell migration and invasion.Besides,the animal experiments exhibited that the nude mice inoculated with NSCLC cells by silencing p300,STAT3 or MMP19 gene plus IL-17 treatment,the nodule number,and MMP19,Ack-STAT3,or p-STAT3 production in the lung metastatic nodules were all alleviated.Collectively,these outcomes uncover that IL-17-triggered NSCLC metastasis involves up-regulating MMP19 expression via the interaction of STAT3-K631 acetylation by p300 and its Y705-phosphorylation,which provides a new mechanistic insight and potential strategy for NSCLC metastasis and therapy.
基金supported by the National Natural Science Foundation of China,No.82104795 (to RH)。
文摘Runx2 is a major regulator of osteoblast differentiation and function;however,the role of Runx2 in peripheral nerve repair is unclea r.Here,we analyzed Runx2expression following injury and found that it was specifically up-regulated in Schwann cells.Furthermore,using Schwann cell-specific Runx2 knocko ut mice,we studied peripheral nerve development and regeneration and found that multiple steps in the regeneration process following sciatic nerve injury were Runx2-dependent.Changes observed in Runx2 knoc kout mice include increased prolife ration of Schwann cells,impaired Schwann cell migration and axonal regrowth,reduced re-myelination of axo ns,and a block in macrophage clearance in the late stage of regeneration.Taken together,our findings indicate that Runx2 is a key regulator of Schwann cell plasticity,and therefore peripheral nerve repair.Thus,our study shows that Runx2 plays a major role in Schwann cell migration,re-myelination,and peripheral nerve functional recovery following injury.
基金supported by Hebei Province Key Research and Development Program(19277770D)Natural Science Foundation of Hebei Province(H2018423026)+2 种基金the Foundation of Health and Family Planning Commission of Hebei(2018068620180688)Fund of Hebei Administration of Traditional Chinese Medicine(2023020).
文摘Collective cancer cell migration(CCCM)and epithelial-to-mesenchymal transition(EMT)play key roles in metastasis.This study reports that the colorectal carcinoma cell line LIM1863 is useful for the study of CCCM and EMT.Methods:Hematoxylin and eosin staining,scanning electron microscopy,transmission electron microscopy,and western blot analysis were performed.Results:LIM1863 automatically grew as spheroids in suspension and had important typical epithelial properties,including several layers of cells arranged around a central lumen,apical-basal polarity,and types of cell-cell junctions.Treatment with a combination of both TGF beta 1 and TNF alpha induced definite and distinct EMT,a spheroid changing phenotype to form a monolayer high-confluent patch without lumen,without polarity.Spontaneous CCCM occurred in spheroids.Flat EMT cells adhered to the base of a dish,exhibited persistent movement as a cluster of cells,and then shed,resulting in a cluster.All cells from one cluster undergoing CCCM died.Otherwise,all cells undergoing EMT disappeared and almost all cells located in the cell reservoir survived and proliferated.Conclusion:LIM1863 is an excellent cell line to study CCCM and EMT.The group of heterogeneous cells undergoing CCCM behaves like a supracellular unit.
基金supported by the National Natural Science Foundation of China(Nos.32102786,32270555).
文摘As a pathway that plays a role in nutrient absorption,anabolic response,cell growth and survival,the important role of AKT/mTOR in tumorigenesis has also come to light.For cancer patients,most deaths are caused by the growth of metastatic tumors outside the primary focus.Therefore,migration and invasion in the late stage of tumor progression are the main unresolved issues in the study of tumor pathogenesis,and AKT/mTOR has been found to participate in the migration and invasion of cancer cells,which means that the study of this pathway may contribute to a solution for the problem.Because of its extensive and complex functions in the organism,this pathway can be regulated by a variety of different signals in the body,and then realize its function through different downstream signal molecules.This article reviews the proteins that can indirectly affect this pathway by regulating the common upstream signaling molecules of this pathway,and the proteins that can directly affect the level of phosphorylation of AKT/mTOR in cancer cells.We also review the proteins that can co-regulate this pathway and its downstream pathways.Through this study,we hope to gain a deeper understanding of the regulatory mechanism of the AKT/mTOR pathway in cancer cells,in hopes of finding effective and harmless cancer treatment targets in the future.
基金supported by the National Natural Science Foundation of China,Nos.81772094(to ZBC),81974289(to ZBC)the Key Research and Development Program of Hubei Province,No.2020BCB031(to ZBC)Natural Science Foundation of Hubei Province,No.2020CFB433(to YTL).
文摘Circular RNAs(circRNAs)play a vital role in diabetic peripheral neuropathy.However,their expression and function in Schwann cells in individuals with diabetic peripheral neuropathy remain poorly understood.Here,we performed protein profiling and circRNA sequencing of sural nerves in patients with diabetic peripheral neuropathy and controls.Protein profiling revealed 265 differentially expressed proteins in the diabetic peripheral neuropathy group.Gene Ontology indicated that differentially expressed proteins were mainly enriched in myelination and mitochondrial oxidative phosphorylation.A real-time polymerase chain reaction assay performed to validate the circRNA sequencing results yielded 11 differentially expressed circRNAs.circ_0002538 was markedly downregulated in patients with diabetic peripheral neuropathy.Further in vitro experiments showed that overexpression of circ_0002538 promoted the migration of Schwann cells by upregulating plasmolipin(PLLP)expression.Moreover,overexpression of circ_0002538 in the sciatic nerve in a streptozotocin-induced mouse model of diabetic peripheral neuropathy alleviated demyelination and improved sciatic nerve function.The results of a mechanistic experiment showed that circ_0002538 promotes PLLP expression by sponging miR-138-5p,while a lack of circ_0002538 led to a PLLP deficiency that further suppressed Schwann cell migration.These findings suggest that the circ_0002538/miR-138-5p/PLLP axis can promote the migration of Schwann cells in diabetic peripheral neuropathy patients,improving myelin sheath structure and nerve function.Thus,this axis is a potential target for therapeutic treatment of diabetic peripheral neuropathy.
基金supported by grants from Shenzhen Key Medical Subject(No.SZXK023)Shenzhen“SanMing”Project of Medicine(No.SZSM201612092)+3 种基金Shenzhen Research and Development Projects(No.JCYJ20170307111755218)Guangdong Basic and Applied Basic Research Foundation(No.2019A1515011290)National Key Research and Development Program of China(No.2016YFC1102103)China Postdoctoral Science Foundation(No.2020M672756)
文摘Interconnectivity is the key characteristic of bone tissue engineering scaffold modulating cell migration,blood vessels invasion and transport of nutrient and waste.However,efforts and understanding of the interconnectivity of porous Mg is limited due to the diverse architectures of pore struts and pore size distribution of Mg scaffold systems.In this work,biomimetic hierarchical porous Mg scaffolds with tailored interconnectivity as well as pore size distribution were prepared by template replication of infiltration casting.Mg scaffold with better interconnectivity showed lower mechanical strength.Enlarging interconnected pores would enhance the interconnectivity of the whole scaffold and reduce the change of ion concentration,pH value and osmolality of the degradation microenvironment due to the lower specific surface area.Nevertheless,the degradation rates of five tested Mg scaffolds were no different because of the same geometry of strut unit.Direct cell culture and evaluation of cell density at both sides of four typical Mg scaffolds indicated that cell migration through hierarchical porous Mg scaffolds could be enhanced by not only bigger interconnected pore size but also larger main pore size.In summary,design of interconnectivity in terms of pore size distribution could regulate mechanical strength,microenvironment in cell culture condition and cell migration potential,and beyond that it shows great potential for personalized therapy which could facilitate the regeneration process.
基金supported by Texas Tech University start-up fund and National Natural Science Foundation of China(51709120).
文摘Cell migration is facilitated by the interaction of living cells and their local microenvironment.The local topography is one of the key factors regulating cell migration.Interaction between the surface topography and the cell behaviors is critical to understanding tissue development and regeneration.In this study,a dynamic mask photolithography technique has been utilized to fabricate a surface with graded micropillars.It has been demonstrated that the cells have been successfully guided to migrate from the sparse zone to the dense zone.The cell polarization angle has been characterized in both sparse zone and the dense zone.Compared to the dense zone,the cells in the sparse zone are more aligned along the direction of the micropillar spacing gradient,which enables the guided cell migration.Moreover,the effects of the micropillar spacing gradient,micropillar diameter,and micropillar height have been investigated in terms of the cell migration speed and cell spreading area.Finally,two issues significantly affecting the cell migration have been discussed:trapped cells between the micropillars and cell clusters.
基金Project supported by the National Natural Science Foundation of China(Grant Nos.11974066,11674043,11675134,and 11874310)the Natural Science Foundation of Chongqing,China(Grant Nos.cstc2019jcyj-msxmX0477 and cstc2018jcyjA3679)the Capital Health Development Research Project,China(Grant No.2020-2-2072).
文摘Cell migration in anisotropic microenvironment plays an important role in the development of normal tissues and organs as well as neoplasm progression,e.g.,osteogenic differentiation of embryonic stem cells was facilitated on stiffer substrates,indicating that the mechanical signals greatly affect both early and terminal differentiation of embryonic stem cells.However,the effect of anisotropy on cell migration dynamics,in particular,in terms of acceleration profiles which is important for recognizing dynamics modes of cell migration and analyzing the regulation mechanisms of microenvironment in mechanical signal transmission,has not been systematically investigated.In this work,we firstly rigorously investigate and quantify the differences between persistent random walk and anisotropic persistent random walk models based on the analysis of cell migration trajectories and velocity auto-covariance function,both qualitatively and quantitatively.Secondly,we introduce the concepts of positive and negative anisotropy based on the motility parameters to study the effect of anisotropy on acceleration profiles,especially the nonlinear decrease and non-monotonic behaviors.We particularly elaborate and discuss the mechanisms,and physical insights of non-monotonic behaviors in the case of positive anisotropy,focusing on the force exerted on migrating cells.Finally,we analyze two types of in vitro cell migration experiments and verify the universality of nonlinear decrease and the consistence of non-monotonic behaviors with numerical results.We conclude that the anisotropy of microenvironment is the cause of the non-monotonic and nonlinear dynamics,and the anisotropic persistent random walk can be as a suitable tool to analyze in vitro cell migration with different combinations of motility parameters.Our analysis provides new insights into the dynamics of cell migration in complex microenvironment,which also has implications in tissue engineering and cancer research.
基金This work is supported by a grant from National Institutes of Health(Grant No.SC2GM112575)a grant from the John L.Santikos Charitable Foundation of the San Antonio Area Foundation.
文摘Cell migration plays a significant role in many biological activities,yet the physical mechanisms of cell migration are still not well understood.In this study,a continuum physics-based epithelial monolayer model including the intercellular interaction was employed to study the cell migration behavior in a confluent epithelial monolayer at constant cell density.The epithelial cell was modeled as isotropic elastic material.Through finite element simulation,the results revealed that themotile cellwas subjected to higher stress than the other jammed cells during the migration process.Cell stiffness was implied to play a significant role in epithelial cell migration behavior.Higher stiffness results in smaller displacement and lower migration speed.
基金This work is supported by a grant from National Institutes of Health(Grant No.SC2GM112575)a grant from the John L.Santikos Charitable Foundation of the San Antonio Area Foundation.
文摘The collective cell migration behavior on a substrate was studied using RKPM meshfree method.The cells were modeled as nematic liquid crystal with hyperelastic cell nucleus.The cell-substrate and cell-cell interactions were modeled by coarse-grained potential forces.Through this study,the pulling and pushing phenomenon during collective cell migration process was observed and it was found that the individual cell mobility significantly influenced the collective cell migratory behavior.More self-propelled cells are in the system along the same direction,the faster the collective group migrates toward coordinated direction.The parametric study on cell-cell adhesion strength indicated that as the adhesion strength increases,the collective cell migration speed increases.It also showed that the mechanical stress in leader cell is higher than stress in follower cells.
基金This research was funded by the Israel Cancer Association,Grant No.20190028 and Ariel University.
文摘The link of the metazoan nucleus to the actin cytoskeleton is highly important for actin polymerization and migration of multiple cell types as well as for mechanotransduction and even affects the cellular transcriptome.Several mechanisms of organization of actin filaments next to the nuclear envelope have been identified.Among these mechanisms the most studied one is the Linker of nucleoskeleton and cytoskeleton(LINC)complex-dependent perinuclear actin organization.However,recently additional mechanisms have been identified:an Actin-related protein-2/3(Arp2/3)-dependent perinuclear actin polymerization during migration of dendritic cells and a perinuclear actin rim that is formed in response to external force application or migration cues.In parallel,there are also reports on cancer cells that migrate in a LINC complex independent manner and on cancers with reduced expression of the LINC complex components.Thus,suggesting that LINC complex independent migration may be associated with tumour formation.
基金This work was supported by Nanjing Municipal Science and Technology Bureau International Joint Research and Development(No.201911041)Science and Technology Development Foundation of Nanjing Medical University(No.NMUB2018327)Social Development project of Jiangsu Province(No.BE2020623).
文摘Abstract Meniscus injury is a common disease in clinic.If it was not treated in time,it leads to osteoarthritis which brings unbearable pain and heavy economic burden to the patients.At present,meniscectomy and meniscus suture are widely used in the treatment for meniscus injury.Nevertheless,It is not ideal for poor self-healing ability of meniscus.The recruitment of endogenous stem cells is an attractive option for wounded meniscus healing.Fully reduced high-mobility group box 1 protein(HMGB1)can accelerate the regeneration of multiple tissues by endogenous stem cell activation,migration and differentiation.Kartogenin(KGN)has shown to induce the chondrogenesis of the stem cells.However,no study has explored such effects of HMGB1 and KGN in wounded meniscus healing.Therefore,in order to improve the regeneration of meniscus,we intend to use a novel bioactive microsphere which was developed by combining fully reduced high mobility group box 1(frHMGB1)and kartogenin(KGN)with alginate gel which slowly release high concentrations of HMGB1 and KGN to activate rat bone marrow stem cells(BMSCs)and promote cell proliferation.The results showed that this HMGB1–KGN microsphere released and kept high concentrations of HMGB1 and KGN in the wound area for more than 2 weeks.In vitro experimental results showed that the HMGB1–KGN microsphere can promote cell proliferation via recruiting rat bone marrow stem cells(BMSCs)and activating the BMSCs from G_(0) to G_(Alert) stage as evidenced by cell migration testing and 5-bromo-2′-deoxyuridine(BrdU)incorporation assay.In vivo results indicated that this HMGB-KGN microsphere can recruit GFP-labeled BMSCs from tail vein to wounded meniscus and induce these GFP-labeled BMSCs to differentiate into chondrocytes.Our results demonstrated that the HMGB1–KGN-containing bioactive microsphere induced cell migration in vitro and recruited the cells to wound area to promote wounded rat meniscus healing in vivo.
基金funds,including the National Natural Science Foundation of China(Nos.81972331,81972751,81572374)The National Key Research and Development Program of China(2017YFC1308900)+2 种基金Technological Special Project of Liaoning Province of China(2019020176-JH1/103)Science and Technology Plan Project of Liaoning Province(NO.2013225585)The General Projects of Liaoning Province Colleges and Universities(LFWK201706).
文摘Distal gastric cancer(DGC)is a subgroup of gastric cancer(GC),which has different molecular characteristics from proximal gastric cancer(PGC).These differences result in different overall survival(OS)rates;however,data pertaining to the survival rate in PGC or DGC are contradictory.This suggests that the location of GC is not the unique cause of the different survival rates,while the molecular characteristics might be more important factors determining the prognosis of DGC.Therefore,the aim of this study was to discover key prognostic factors in DGC using bioinformatic methods and to explore the potential molecular mechanism.The Cancer Genome Atlas(TCGA)public database was employed to screen data relating to DGC,and we conducted a weighted gene co-expression network analysis(WGCNA)on DGC patient samples to establish co-expression modules.High-weight genes(hub genes)in a dominant color module were identified.In vitro experiments and gene set enrichment analyses(GSEA)were carried out to elucidate the potential molecular mechanism.In this study,139 DGC samples were enrolled to perform a co-expression analysis.According to the correlation between gene modules and clinical characteristics,the royal blue module related to stage M of DGC was screened,and a survival analysis was conducted to show that highcoagulation-factor V(F5)expression was related to the short OS of patients with GC.In vitro experiments confirmed that F5 could promote the migration of GC cells.GSEA suggested that F5 might have affected the prognosis of GC by modulating the activities of the Wnt and/or the TGF-βsignaling pathways.Our results indicated that high F5 expression predicts poor prognosis of patients with DGC,and it functions probably by promoting cell migration through the Wnt and/or the TGF-βsignaling pathways.
基金The funding for the author’s research has been supported by the National Eye Institute—NIH,The Michigan Eye Bank,Midwest Eye Bank,Eversight,the Research Excellence Fund of the Center for Biomedical Sciences,Oakland University and the Department of Biological Sciences.
文摘Much of our understanding of the events which underlie cell migration has been derived from studies of cells intissue culture. One of the components that mediates this process is the dynamic actin-based microfilament system that canreorganize itself into so-called stress fibers that are considered essential components for cell motility. In contrast, relativelyfew studies have investigated cell movement along an extracellular matrix (ECM) which is known to influence both cellularorganization and behavior. This opinion/viewpoint article briefly reviews cell migration during corneal endothelial woundrepair along the tissue’s natural basement membrane, Descemet’s membrane. Because the tissue exists as a cell monolayer itaffords one an opportunity to readily explore the effect of cell/matrix influences on cell motility. As such, cell movementalong this substrate differs somewhat from that found in vitro and migrating endothelial cells also demonstrate an abilityto move along the ECM without the benefit of having an organized actin cytoskeleton.
文摘Migration of dendritic cells (DCs) into tissues and secondary lymphoid organs plays a crucial role in the initiation of innate and adaptive immunity. In this article, we show that cyclosporin A (CsA) impairs the migration of DCs both in vitro and in vivo. Exposure of DCs to clinical concentrations of CsA neither induces apoptosis nor alters development but does impair cytokine secretion, chemokine receptor expression, and migration. In vitro, CsA impairs the migration of mouse bone marrow-derived DCs toward macrophage inflammatory protein-3beta (MIP-3beta) and induces them to retain responsiveness to MIP-1alpha after lipopolysaccharide (LPS)-stimulated DC maturation, while in vivo administration of CsA inhibits the migration of DCs out of skin and into the secondary lymphoid organs. CsA impairs chemokine receptor and cyclooxygenase-2 (COX-2) expression normally triggered in LPS-stimulated DCs; administration of exogenous prostaglandin E2 (PGE2) reverses the effects of CsA on chemokine receptor expression and DC migration. Inhibition of nuclear factor-kappaB (NF-κB) and mitogen-activated protein kinase (MAPK) pathway signaling by CsA may be responsible for the CsA-mediated effects on the regulation of chemokine receptor and cyclooxygenase-2 (COX-2) expression. Impairment of DC migration due to inhibition of PGE2 production and regulation of chemokine receptor expression may contribute, in part, to CsA-mediated immunosuppression.
文摘Background To investigate the effects and mechanisms of cellular repressor of ElA stimulated genes (CREG) on endothelial cell(EC) migration.Methods vascular endothelial cells(VE),CREG overexpression VEs, CREG suppression VEs and VEs transfected with CREG gene modified adenovirus(Ad-CREG) were cultured with dulbecco’s modified eagle’s medium contained 10%fetal calf serum. Western blot was used to detect the protein level of CREG and integrin-linked kinase(ILK) in the four kind ECs.Tran-swell migration model was applied to compare the migration cell number of the four kind ECs.Two kinds of ILK mutant plasmids;PCXN2-flag-ILK wt-IRES-GFP(wild-type ILK)and PCXN2-flag-ILK p-parvin-IRES-GFP(P-parvin-binding mutant) were used to transfect VS and VE respectively,then the two kind transfection ECs were named as VS-wtILK and VE-P -parvin which were selected by G418(600ng/ml)for 2 weeks;Transwell migration model was applied to compare migration capability before and after ILK plasmids transfecting VE and VS.Results Western blot analysis showed that CREG overexpression promoted ILK expression in ECs,on the contrary,ILK expression was down-regulated in CREG silent ECs(P【0.05).Further more,ILK expression was up-regulated obviously in VE transfected with Ad-CREG(P【 0.05);Transwell migration model showed that EC’s migration capability was positively correlated with the expression level of CREG in EC,that is,CREG overexpression induced VE migration and CREG silent suppressed VE migration, moreover,Ad-CREG transfecting VE showed better migration capability accompanied with CREG expression increase by transwell migration model(P【0.05).In order to know the relationship between ILK expression and cell migration,we obtained stable transfection cell strains of VS-wtILK and VE-Pparvin, transwell migration model demonstrated that VS-wtILK remarkably corrected the poor migration capability of VS(P【 0.01),butβ-parvin combining site mutation in ILK genes inhibited VE migration markedly(P【0.01).Conclusions ILKp -parvin signal pathway mediated vascular endothelial cell migration induced by CREG.
基金Supported by Grants-in-Aid for Young Scientists(B)No.22791295,No.23791557 and No.24791440Grant-in-Aid for Scientific Research(C)from the Japan Society for the Promotion of Science,No.22591464 and No.24591957
文摘AIM:To investigate the role of claudin 1 in the regulation of genes involved in cell migration and tumor necrosis factor alpha(TNF-α)-induced gene expression in human gastric adenocarcinoma cells.METHODS:Knockdown experiments were conducted with claudin 1 small interfering RNA(si RNA),and theeffects on the cell cycle,apoptosis,migration and invasion were analyzed in human gastric adenocarcinoma MKN28 cells.The gene expression profiles of cells were analyzed by microarray and bioinformatics.RESULTS:The knockdown of claudin 1 significantly inhibited cell proliferation,migration and invasion,and increased apoptosis.Microarray analysis identified 245genes whose expression levels were altered by the knockdown of claudin 1.Pathway analysis showed that the top-ranked molecular and cellular function was the cellular movement related pathway,which involved MMP7,TNF-SF10,TGFBR1,and CCL2.Furthermore,TNFand nuclear frctor-κB were the top-ranked upstream regulators related to claudin 1.TNF-αtreatment increased claudin 1 expression and cell migration in MKN28 cells.Microarray analysis indicated that the depletion of claudin1 inhibited 80%of the TNF-α-induced m RNA expression changes.Further,TNF-αdid not enhance cell migration in the claudin 1 si RNA transfected cells.CONCLUSION:These results suggest that claudin 1 is an important messenger that regulates TNF-α-induced gene expression and migration in gastric cancer cells.A deeper understanding of these cellular processes may be helpful in establishing new therapeutic strategies for gastric cancer.
基金Supported by the National Natural Science Foundation of China(81472485)the Project of Six Talent Peaks of Jiangsu Province(WSW-014)Project of Hospital Management Center of Wuxi(YGZXQ1311,YGZ1106,and YGZ1007)
文摘Objective To investigate the effects of microRNA-18a(miR-18a) on migration and invasion of hepatocellular carcinoma(HCC) cells,and its possible mechanism associated with Dicer l. Methods HepG 2 and HepG 2.2.15 cells were transfected with miR-18 a inhibitor using Lipofectamine. Cell invasion was evaluated by transwell invasion assay,and cell migration was detected by transwell migration and wound-healing assays. Moreover,luciferase reporter assay was used to identify whether Dicer expression was regulated by miR-18 a. Real-time RT-PCR and western blot were performed to analyze Dicer 1 expression. In addition,a functional restoration assay was performed to investigate whether miR-18 a promotes HCC cell migration and invasion by directly targeting Dicer 1. Results miR-18 a inhibitor can suppress the migration and invasion of HCC cells. Furthermore,suppression of Dicer l expression by small interfering RNA essentially abolished the inhibition of cell migration and invasion induced by miR-18 a inhibitor,restorating these activities to levels similar to the parental HCC cells. Interestingly,suppression of miR-18 a in HCC cells resulted in enhanced expression of Dicer l. In addition,the results of a luciferase assay demonstrated targeted regulation of Dicer l by miR-18 a. Conclusion Our findings suggest that miR-18 a promotes migration and invasion of HCC cells by inhibiting Dicer l expression.
基金financially supported by the National Natural Science Foundation of China ( 11532009, 11602191,21775117)the General Financial Grant from the China Postdoctoral Science Foundation ( 2016M592773)the High Level Returned Overseas Students Foundation ( [2018]642)
文摘The migration mode transition of cancer cell enhances its invasive capability and the drug resistance,where physical confinement of cell microenvironment has been revealed to induce the mesenchymal-amoeboid transition(MAT).However,most existing studies are performed in PDMS microchannels,of which the stiffness is much higher than that of most mammalian tissues.Therefore,the amoeboid migration transition observed in these studies is actually induced by the synergistic effect of matrix stiffness and confinement.Since the stiffness of cell microenvironment has been reported to influence the cell migration in 2D substrate,the decoupling of stiffness and confinement effects is thus in need for elucidating the underlying mechanism of MAT.However,it is technically challenging to construct microchannels with physiologically relevant stiffness and channel size,where existing microchannel platforms with physiological relevance stiffness are all with>10μm channel width.Such size is too wide to mimic the physical confinement that migrating cancer cells confront in vivo,and also larger than the width of PDMS channel,in which the MAT of cancer cell was observed.Therefore,an in vitro cell migration platform,which could mimic both stiffness and confinement of the native physical microenvironment during cancer metastasis,could profoundly contribute to researches on cancer cell migration and cellular mechanotransduction.In this paper,we overcome the limitations of engineering soft materials in microscale by combining the collagen-alginate hydrogel with photolithography.This enables us to improve the accuracy of molded microchannel,and thus successfully construct a 3D microchannel platform,which matches the stiffness and width ranges of native environmental confinement that migrating cancer cells confront in vivo.The stiffness(0.3~20 kPa),confinement(channel width:3.5~14μm)and the adhesion ligand density of the microchannel can be tuned independently.Interestingly,using this platform,we observed that the migration speed of cancer cell is influenced by the synergistic effect of channel stiffness and width,and the increasing stiffness reverses the effect of channel width on the migration speed of cancer cells.In addition,MAT has a strong correlation with the channel stiffness.These findings make us reconsider the widely accepted hypothesis:physical confinement can induce MAT.Actually,this transition can only occur in stiff confined microenvironment not in soft one.For soft microchannels,the compliance of the channel walls could cause little cell/nucleus deformation,and the MAT could not be induced.To further investigate the mechanism of MAT,we developed a computational model to simulate the effect of nucleus deformation on MAT.With the model,we found that deforming the cell nuclear by decreasing the nucleus stiffness will reduce the cellmigration speed.This implies that nuclear stiffness plays an important role in the regulation of cancer migration speed and thus MAT in microchannels.The effect of channel stiffness on MAT and migration speed as observed in our experiment could partially explain previous findings reported in the literature,where the increasing matrix stiffness of tumor microenvironment promotes cancer metastasis.Our observations thus highlight the critical role of cell nuclear deformation not only in MAT,but also in regulating cellular mechanotransduction and cell-ECM interactions.This developed platform is capable of mimicking the native physical microenvironment during metastasis,providing a powerful tool for high-throughput screening applications and investigating the interaction between cancer migration and biophysical microenvironment.
基金We thank Drs Chenbing Guan and Kui Cui （Institute of Neuroscience, Shanghai Institute for Biological Science, China） for technical support in setting up the single-cell migration assay, and Dr Qian Hu （Institute of Neuroscience, Shanghai Institute for Biological Science, China） for microscopic imaging. This study was supported by the National Key Basic Research Program （2006CB500702）, Ministry of Science and Technology of China （2007CB947100）, National Natural Science Foundation of China （30530240 and 30770657）, Program for Changdiang Scholars and Innovative Research Teams in Universities （IRT0528）, and Shanghai Metropolitan Fund for Research and Development （07DJ14005）.
