Aging is the leading risk factor for Alzheimer’s disease and other neurodegenerative diseases. We now understand that a breakdown in the neuronal cytoskeleton, mainly underpinned by protein modifications leading to t...Aging is the leading risk factor for Alzheimer’s disease and other neurodegenerative diseases. We now understand that a breakdown in the neuronal cytoskeleton, mainly underpinned by protein modifications leading to the destabilization of microtubules, is central to the pathogenesis of Alzheimer’s disease. This is accompanied by morphological defects across the somatodendritic compartment, axon, and synapse. However, knowledge of what occurs to the microtubule cytoskeleton and morphology of the neuron during physiological aging is comparatively poor. Several recent studies have suggested that there is an age-related increase in the phosphorylation of the key microtubule stabilizing protein tau, a modification, which is known to destabilize the cytoskeleton in Alzheimer’s disease. This indicates that the cytoskeleton and potentially other neuronal structures reliant on the cytoskeleton become functionally compromised during normal physiological aging. The current literature shows age-related reductions in synaptic spine density and shifts in synaptic spine conformation which might explain age-related synaptic functional deficits. However, knowledge of what occurs to the microtubular and actin cytoskeleton, with increasing age is extremely limited. When considering the somatodendritic compartment, a regression in dendrites and loss of dendritic length and volume is reported whilst a reduction in soma volume/size is often seen. However, research into cytoskeletal change is limited to a handful of studies demonstrating reductions in and mislocalizations of microtubule-associated proteins with just one study directly exploring the integrity of the microtubules. In the axon, an increase in axonal diameter and age-related appearance of swellings is reported but like the dendrites, just one study investigates the microtubules directly with others reporting loss or mislocalization of microtubule-associated proteins. Though these are the general trends reported, there are clear disparities between model organisms and brain regions that are worthy of further investigation. Additionally, longitudinal studies of neuronal/cytoskeletal aging should also investigate whether these age-related changes contribute not just to vulnerability to disease but also to the decline in nervous system function and behavioral output that all organisms experience. This will highlight the utility, if any, of cytoskeletal fortification for the promotion of healthy neuronal aging and potential protection against age-related neurodegenerative disease. This review seeks to summarize what is currently known about the physiological aging of the neuron and microtubular cytoskeleton in the hope of uncovering mechanisms underpinning age-related risk to disease.展开更多
BACKGROUND Glioma is one of the most common intracranial tumors,characterized by invasive growth and poor prognosis.Actin cytoskeletal rearrangement is an essential event of tumor cell migration.The actin dynamics-rel...BACKGROUND Glioma is one of the most common intracranial tumors,characterized by invasive growth and poor prognosis.Actin cytoskeletal rearrangement is an essential event of tumor cell migration.The actin dynamics-related protein scinderin(SCIN)has been reported to be closely related to tumor cell migration and invasion in several cancers.AIM To investigate the role and mechanism of SCIN in glioma.METHODS The expression and clinical significance of SCIN in glioma were analyzed based on public databases.SCIN expression was examined using real-time quantitative polymerase chain reaction and Western blotting.Gene silencing was performed using short hairpin RNA transfection.Cell viability,migration,and invasion were assessed using cell counting kit 8 assay,wound healing,and Matrigel invasion assays,respectively.F-actin cytoskeleton organization was assessed using F-actin staining.RESULTS SCIN expression was significantly elevated in glioma,and high levels of SCIN were associated with advanced tumor grade and wild-type isocitrate dehydrogenase.Furthermore,SCIN-deficient cells exhibited decreased proliferation,migration,and invasion in U87 and U251 cells.Moreover,knockdown of SCIN inhibited the RhoA/focal adhesion kinase(FAK)signaling to promote F-actin depolymerization in U87 and U251 cells.CONCLUSION SCIN modulates the actin cytoskeleton via activating RhoA/FAK signaling,thereby promoting the migration and invasion of glioma cells.This study identified the cancer-promoting effect of SCIN and provided a potential therapeutic target for the treatment of glioma.展开更多
Anticancer drugs are one of the most direct means of cancer therapy.However,the various cancer progressions hamper the development and discovery of anticancer drugs.In fact,the mechanical properties of the tumor cytos...Anticancer drugs are one of the most direct means of cancer therapy.However,the various cancer progressions hamper the development and discovery of anticancer drugs.In fact,the mechanical properties of the tumor cytoskeleton are extremely vital for any phase of cancer,especially in tumor invasion and metastasis.However,in the current category of anticancer drugs,the cytoskeleton-targeting drugs are limited and their role in tumor progression is unclear.Here,we present the mechanical characteristics of tumor stiffness that are tightly regulated by the cancer cytoskeleton,including actin filaments and microtubules during tumor initiation,growth and metastasis,and review the natural drugs that target the cancer cytoskeleton.We define cytoskeleton dynamics as target mechanisms for anticancer drugs and summarize the plant,microbial and marine sources of natural products.Furthermore,this paper also provides a material pathway to study active tumor mechanics,and introduces the unique advantages and future application potential of tumor cytoskeleton-targeting drugs in clinical use.The material approaches to active cancer mechanics are supplied in this review.We aim to promote the development of anticancer drugs that target tumor mechanics by using those material approaches and finding their pharmacological application.展开更多
Background:In myocardial ischemia,hypoxia leads to destruction of the cytoskeleton,and especially the imbalance of microtubule polymerization-depolymerization,which seriously affects the structure and function of card...Background:In myocardial ischemia,hypoxia leads to destruction of the cytoskeleton,and especially the imbalance of microtubule polymerization-depolymerization,which seriously affects the structure and function of cardiomyocytes.We previously showed that a Yiqi Huoxue Decoction (YQHX) improves mitochondrial function and decreases anti-oxidative effects in hypoxia-induced H9c2 cell injury.Therefore,in this study we investigated whether YQHX protects against hypoxia-induced damage by decreasing damage to the cardiac cytoskeleton.Methods:After reaching 70%-80% confluence,H9c2 cells were synchronized in serum-free Dulbecco's Modified Eagle Medium for 6 hours,then divided into control,model,and YQHX (100,200,400 μg/mL) groups,which were then grown in a hypoxic atmosphere for 12 hours.Cardiac cell viability was assessed using an xCELLigence system.The levels of lactate dehydrogenase,maleic dialdehyde,and superoxide dismutase in H9c2 cell supernatants were measured.Hoechst 33258 staining was employed to observe cardiac cell apoptosis.Confocal microscopy,immunofluorescence,and western blot analysis were performed to evaluate the protective effects of the YQHX against hypoxia-induced injury in the H9c2 cell line.Results:Cells that were pretreated with YQHX were more able to maintain their microtubule structure in the early stages of hypoxia and had better myocardial fitness in response to hypoxia compared with cells that were not pretreated.However,hypoxia-induced upregulation of α-tubulin and β-tubulin expression antagonized the protective effect of YQHX (100 μg/mL).In addition,YQHX (100 μg/mL) treatment significantly upregulated MAP4 protein expression (P =.003) and downregulated p-AMPKα protein expression (P <.001) compared with the model group.Conclusion:The results indicate that YQHX plays a role in protecting against oxidative stress injury and apoptosis in H9c2 cells.Notably,our results suggested that the YQHX could mitigate the damage to the cardiac cytoskeleton and the dysregulation of AMPK-related protein signaling pathways that are induced by hypoxia.展开更多
Objective Fumonisin B1(FB1)is an important mycotoxin in nature worldwide.The biomechanical properties of cells are closely related to their structure and function,and the cytoskeleton is the structural and functional ...Objective Fumonisin B1(FB1)is an important mycotoxin in nature worldwide.The biomechanical properties of cells are closely related to their structure and function,and the cytoskeleton is the structural and functional basis of cells motility,and therefore,from a biomechanical point of view,the purpose of this study is to investigate the effects of FB1 on the biomechanical properties,migration capacity and cytoskeletal structure of human umbilical vein endothelial cells(HUVECs),which may lay an experimental foundation for further exploration of the toxicity mechanism of fumonisin.Methods HUVECs were cultured and treated with different concentrations of FB1.Then,CCK-8 kit was used to detect the effect of FB1 on the survival rate.The osmotic fragility of the cells was measured after treatment with different osmotic pressures for30 min.The cell membrane fluidity was measured by fluorescence polarization method.The cell electrophoretic mobility was measured by cell electrophoretic apparatus.The migration capacity of the cells was observed by scratch repair assay.The changes of reactive oxygen species and cytoskeletal structure were observed by confocal laser scanning microscopy.Finally,the mRNA and protein relative expression levels of cytoskeletal binding proteins were detected by real-time PCR,Western blotting and confocal laser scanning.Results The results of CCK-8 showed that FB1 could significantly inhibit the proliferation of HUVECs in a dose-and time-dependent manner.After treatment of HUVECs with FB1,the hypotonic resistance of the cell,cell surface charge,cell membrane fluidity and migration capacity were all weakened,while reactive oxygen species were significantly increased and the cytoskeletal structure was significantly reorganized.Furthermore,RTPCR results showed that the mRNA relative expression levels of cytoskeletal binding proteins,exception of actin,were down-regulated after treated with FB1.Besides,Western blotting and statistical analysis based on fluorescence intensity of laser confocal microscopy confirmed theses changes in protein level.Conclusions FB1 can significantly affect the biomechanical properties and motility of HUVECs,which may be directly correlated to the remodel of F-actin cytoskeleton,as well as the relative expression changes of cytoskeletal binding proteins.It is significant for further exploring the toxicity mechanism of fumonisin.展开更多
Background Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.The dynamics of cytoskeleton plays crucial regulated roles o...Background Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.The dynamics of cytoskeleton plays crucial regulated roles on DCs’immune functions and biophysical properties.Several evidences show that tumor-derived suppressive cytokines deteriorate DCs’immune functions through remodeling their F-actin cytoskeleton.But the underlying mechanism is still elusive.Tropomodulin1(Tmod1),a cytoskeleton-binding protein,regulates and stabilizes actin filaments lengths and cytoskeleton architecture,which involves in the regulations of the morphology,formation of neural dendrites and biophysical properties of cells.Our previous studies found that mature DCs(mDCs)had a higher expression of Tmod1 than immature DCs(imDCs). Therefore,it’s hypothesized that Tmod1 maybe involve in the modification of DCs’functions.Objective The aim of the study is to investigate the effects of Tmodl on the immune functions and biophysical properties of DCs and the underlying mechanisms in order to further understand the biological behaviors of DCs.Methods Bone marrow-derived cells were harvested from wild type(C57BL/6 J)mice and Tmod1 knockout mice(Tmod1 overexpressing transgenic(TOT)/Tmod1-/-)and differentiated to immature dendritic cells(imDCs)by rmGM-CSF and rmIL-4.imDCs were then matured by lipopolysaccharides(LPS)treatment.The expressions of the surface markers in DCs,including CD80,CD86,CD40,MHC-Ⅱand CCR7,were detected by flow cytometry,Western blot and qRT-PCR.The inflammation cytokines such as IL-6,IFN-γ,IFN-βand IL-10 were also detected by flow cytometry.The immune functions and the biophysical properties of DCs were compared between the wild type and Tmod1 knockout mice.The F-actin content and dendritic pseudopodia of these two kinds of DCs were detected by flow cytometry and laser scanning confocal microscope respectively.Finally,we detected the MyD88 dependent and independent signaling pathway to discover the molecular mechanisms.Results We found that Tmod1-deficient mDCs showed deficient antigen-presenting ability and they failed to express enough MHC-Ⅱ,co-stimulated molecules(CD80/86,CD40)and CCR7 on their cell surface.The secretions of the inflammatory cytokines IL-6 and IFN-γwere decreased while the anti-inflammatory cytokines IFN-βand IL-10 were increased in the supernatant of Tmod1-deficient mDCs.As compared to DCs of wild type mice,the migration ability of DCs from Tmod1 knockout mice were dramatically damaged including their free migration and CCL19 mediated chemotaxis migration.However,we found that Tmod1 knockout had no effects on the imDCs’endocytosis ability.Furthermore,Tmod1 knockout DCs showed higher osmotic fragility,lower Young’s modulus,less F-actin content and shorter dendritic pseudopodia.Under LPS stimulation,the phosphorylation level of p65 and p38 were significantly downregulated in Tmod1 knockout mice while the expression of p-IRF3 was upregulated.Conclusions These results indicated that Tmodl knockout leads to deficient antigen-presenting ability and impaired migration of DCs as well as their biophysical properties.The underlying mechanisms are due to the inhibitions of the TLR4-mediated NF-κB and p38 MAPK singling pathway and the activation of the IRF3 signaling pathway,as well as the disturbed reorganization of the F-actin cytoskeleton.Our results provide a new insight on the functions of Tmod1 which can affect the DCs’immune functions and biophysical properties through regulating the TLR4-mediated singling pathways and cytoskeleton remodeling.展开更多
Objective:To examine the role of heat shock protein 90(Hsp90) in the maintenance of actin cytoskeleton in human neuroblastoma tumor cells.Methods:Co-precipitation experiments were performed to examine Hsp90 interactio...Objective:To examine the role of heat shock protein 90(Hsp90) in the maintenance of actin cytoskeleton in human neuroblastoma tumor cells.Methods:Co-precipitation experiments were performed to examine Hsp90 interaction with actin.Hsp90 and actin interactions were evaluated by protein refolding and acto-myosin motility assays.17-(AUylamino)-17- demethoxygeldanamycin(17AAG) induced actin-cytoskeleton re-organization was examined by laser scanning confocal microcopy.Results:It was shown that inhibition of Hsp90 by 17AAC accelerates detergent induced cell lysis of neuroblastoma tumor cells through destabilization of actin cytoskeleton.The in vitro co-precipitation experiments showed that functional but not mutant Hsp90 binds with F-actin.Among biochemical modifications,phopshorylation and oligomerization enhanced Hsp90 binding with F-actin.F-actin binding to Hsp90 interfered with Hsp90 chaperone activity in protein refolding assays,and Hsp90 binding to F-actin interfered with actin motility on myosin coated flow cell.In the combination treatment,17AAG irreversibly augmented the effect of cytochalasin D,an inhibitor of actin polymerization.Conclusions:It can be concluded that Hsp90 binds to F-actin in tumor cells and maintains the cellular integrity. The results display a novel element of Hsp90 inhibition in destabilizing the actin cytoskeleton of tumor cells,therefore suggest that 17AAG combination with cytoskeletal disruptor may be effective in combating cancer.展开更多
AIM:To investigate the effects of dexamethasone(DEX) and 1-(5-isoquinolinesulfonyl)-homopiperazine(HA1077) on actin cytoskeleton and β-catenin in cultured human trabecular meshwork(HTM) cells.METHODS: The HTM cells w...AIM:To investigate the effects of dexamethasone(DEX) and 1-(5-isoquinolinesulfonyl)-homopiperazine(HA1077) on actin cytoskeleton and β-catenin in cultured human trabecular meshwork(HTM) cells.METHODS: The HTM cells were separated from human eyeball and cultured in vitro.They were divided into control group,DEX(1×10^(-6)mol/L) group,HA1077(3×10^(-5)mol/L)group,and DEX(1×10^(-6)mol/L) and HA1077(3×10^(-5)mol/L)group.Actin cytoskeleton and β-catenin in HTM cells of the four groups were examined by immunofluorescence and Western blot analyses.RESULTS: In DEX group,there were reorganization of actin cytoskeleton and formation of cross linked actin networks(CLANs),which were partially reversed in DEX and HA1077 group.DEX treatment also induced an increased expression of β-catenin,which was obviously reduced in DEX and HA1077 group.Meanwhile,the cultured HTM cells in HA1077 group had lower expression of β-catenin than that in the control group. CONCLUSION: Our results show that HA1077 can reverse the changes of actin organization and expression of β-catenin induced by DEX in cultured HTM cells,suggesting that HA1077 may play an important role in increasing outflow and reducing intraocular pressure.展开更多
We have demonstrated that the distribution of microtubules (MT), mlcrofilaments (MF) and fibronectin (FN) were diminished, while the gene expression of the calmodulin and c- fos enhanced in the transformed C3H10T1/2 c...We have demonstrated that the distribution of microtubules (MT), mlcrofilaments (MF) and fibronectin (FN) were diminished, while the gene expression of the calmodulin and c- fos enhanced in the transformed C3H10T1/2 cells. After treatment with 1 mM db-cAMP for 1 hour and 2 hours, there was an early and repldly reduced in gene expression of Calmodulin and c-fos respectively. After db-cAMP treatment for 4 -5 days, the number of capping cells of ConA binding decreased significantly and the cell surface microvllll decreased as well. The growth of treated cells was inhibited markedly. By using 4F1 cDNA probe, which is preferentially expressed In G1 phase, we have found that the db- cAMP treated cells were accumulated at G1 phase. Of particular interest is the fact that the distribution of microtubules, mlcrofilaments and fibronectln were recovered after treatment with 1 mM db-cAMP for 6 days. It is suggested that the Inhibition of proliferation, alteration, of phenotype and reco- very of cytoskeleton is展开更多
To evaluate the effects of Hantaan(HT) virus on the cytoskeleton of human endothelial cells(HECs).The human umbilical vein endothelial cells were isolated and cultured in vitro.After confluence,the SR-ll strain of HT ...To evaluate the effects of Hantaan(HT) virus on the cytoskeleton of human endothelial cells(HECs).The human umbilical vein endothelial cells were isolated and cultured in vitro.After confluence,the SR-ll strain of HT virus(100 and,300 TCID_(50)) were inoc展开更多
Focal adhesions are polyproteins linked to extracellular matrix and cytoskeleton,which play an important role in the process of transforming force signals into intracellular chemical signals and subsequently triggerin...Focal adhesions are polyproteins linked to extracellular matrix and cytoskeleton,which play an important role in the process of transforming force signals into intracellular chemical signals and subsequently triggering related physiological or pathological reactions.The cytoskeleton is a network of protein fibers in the cytoplasm,which is composed of microfilaments,microtubules,intermediate filaments,and cross-linked proteins.It is a very important structure for cells to maintain their basic morphology.This review summarizes the process of fluid shear stress transduction mediated by focal adhesion and the key role of the cytoskeleton in this process,which focuses on the focal adhesion and cytoskeleton systems.The important proteins involved in signal transduction in focal adhesion are introduced emphatically.The relationship between focal adhesion and mechanical transduction pathways are discussed.In this review,we discuss the relationship between fluid shear stress and associated diseases such as atherosclerosis,as well as its role in clinical research and drug development.展开更多
Objective: To study the mechanisms of the injury of the cytomembrane and cytoskeleton of cultured myocardial cells exposed to the combined stimulation of hypoxia and burn serum. Methods: The myocardial cells of neonat...Objective: To study the mechanisms of the injury of the cytomembrane and cytoskeleton of cultured myocardial cells exposed to the combined stimulation of hypoxia and burn serum. Methods: The myocardial cells of neonatal rats were cultured in vitro. The changes of the cytoskeleton and cytomembrane of the cultured myocardial cells before and after the combined stimulation of hypoxia and burn serum were observed dynamically with immunohistochemistry, flow cytometry and viscoelasticity determination technique. The deformity of the cultured myocardial cells was determined mathematically. Results: In the early stage after the combined impact of hypoxia and burn serum, the cultured myocardial cells were deformed. The elastic coefficient k1 and k2 and the viscosity coefficient μ of the cytomembrane were significantly decreased (P < 0.05 - 0. 01 ). The fluorescent intensity of the cytoskeleton protein and the number of the intermediate filaments and imcrotubules were markedly decreased. Conclusion: The cytoskeleton is essential to the integrity of the cytomembrane. The damage of the cytoskeleton induces the increase of fragility and decrease of viscoelasticity of the cytomembrane . These changes of the biomechanic characteistics participate directly in the initiation and development myocardial damage.展开更多
The manuscript deals with our studies and experiences in the assessment of cytoskeleton in different cellular models and situations.The immunofluorescent study of several cytoskeletal proteins was relevant in the eval...The manuscript deals with our studies and experiences in the assessment of cytoskeleton in different cellular models and situations.The immunofluorescent study of several cytoskeletal proteins was relevant in the evaluation of a therapy for osteoarthritis,in case of alkaptonuria and in testing the efficacy of docetaxel in neuroblastoma cancer cells leading to apoptosis.A relevant part of our experience focus on the study of cytoskeleton in seminiferous epithelium and spermatozoa,identifying alterations affecting blood-testis barrier after a silver nanoparticle treatment,chromosomal segregation in case of varicocele,sperm motility and diagnosing systematic sperm defects as“Primary ciliary dyskinesia”and“Dysplasia of the fibrous sheath”.The evaluation of cytoskeleton represents a specific and sensitive analysis in establishing the health status of different cells.展开更多
Cytoskeletal remodeling affects the shape,adhesion,and motility of cells.Cytoskeletal dynamics are modulated by matrix proteins,integrins,and several cytokines in the tumor microenvironment.In this scenario,signaling ...Cytoskeletal remodeling affects the shape,adhesion,and motility of cells.Cytoskeletal dynamics are modulated by matrix proteins,integrins,and several cytokines in the tumor microenvironment.In this scenario,signaling is activated by integrins and interferons,which can induce ISG15 gene expression.This gene encodes a ubiquitin-like protein that functions as a protein modifier via the ISGylation system.Furthermore,non-conjugated ISG15 acts as a cytokine-like protein.In this viewpoint,the interplay between free ISG15,protein ISGylation,and cytoskeletal dynamics in the tumor microenvironment is discussed.展开更多
Two different simple cases of plane tensegrity cytoskeleton geometries are presented and investigated in terms of stability. The tensegrity frames are used to model adherent cell cytoskeletal behaviour under the appli...Two different simple cases of plane tensegrity cytoskeleton geometries are presented and investigated in terms of stability. The tensegrity frames are used to model adherent cell cytoskeletal behaviour under the application of plane substrate stretching and describe thoroughly the experimentally observed reorientation phenomenon. Both models comprise two elastic bars (microtubules), four elastic strings (actin filaments) and are attached on an elastic substrate. In the absence of external loading shape stability of the cytoskeleton is dominated by its prestress. Upon application of external loading, the cytoskeleton is reorganized in a new direction such that its total potential energy is rendered a global minimum. Considering linear constitutive relations, yet large deformations, it is revealed that the reorientation phenomenon can be successfully treated as a problem of ma- thematical stability. It is found that apart from the magnitude of contractile prestress and the magnitude of extracellular stretching, the reorientation is strongly shape–dependent as well. Numerical applications not only justify laboratory data reported in literature but such experimental evidence as the concurrent appearance of two distinct and symmetric directions of orientation, indicating the cellular coexistence of phases phenomenon, are clearly detected and incorporated in the proposed mathematical treatment.展开更多
Objective:To observe the effect of lipopolysaccharide(LPS)on cytoskeleton and the effect of Dahuang Lingxianfang on nF-KB/MAPK signaling pathway.Methods:Biliary epithelial cells of each group were stained with photoli...Objective:To observe the effect of lipopolysaccharide(LPS)on cytoskeleton and the effect of Dahuang Lingxianfang on nF-KB/MAPK signaling pathway.Methods:Biliary epithelial cells of each group were stained with photolipin fluorescent staining,and the arrangement of cytoskeleton was observed under laser confocal microscope.Western blotting was used to detect the expression of f-actin.Results:After LPS intervention,the biliary epithelial cells showed nuclear shrinkage or damage,and the skeleton was broken or lumped.The cytoskeleton was partially repaired after the intervention of pathway blocking preparation combined with RHUbarb Lingxianfang.All the other groups had different degree of cytoskeleton fracture.Compared with normal group,the expression of F-actin protein in LPS group was decreased(P<0.05);Compared with LPS group,the expression of F-actin in LPS+TCM group,LPS+PDTC+TCM group,LPS+SB203580+TCM group and LPS+PDTC+SB203580+TCM group was significantly increased(P<0.05);Compared with traditional Chinese medicine group,the expression of F-actin in LPS+PDTC+traditional Chinese medicine group,LPS+SB203580+traditional Chinese medicine group and LPS+PDTC+SB203580+traditional Chinese medicine group had no significant difference(P>0.05).Conclusion:RHUbarb Lingxianfang can restore the sequence of biliary epithelial cytoskeleton and protect its microfilament structure under inflammation,and its mechanism may be related to the regulation of NF-KB/MAPK signaling pathway.展开更多
Cytoskeleton exists in all eukaryotes and is involved in many significant cytobiological processes, especially the movements and developmental changes of plant cells. The cytoskeleton consists of microtubule (MT), mic...Cytoskeleton exists in all eukaryotes and is involved in many significant cytobiological processes, especially the movements and developmental changes of plant cells. The cytoskeleton consists of microtubule (MT), microfilament (MF), and intermediate filament (IF). MT and MF are vital components of plant cytoskeleton. Crosslinking factor acts as a bridge between MF and MT. They play an important role in cellular life process and have always been a hot topic and key point in plant cytobiology, and the IF is a difficult point in this field. In this paper, the latest research on the cytoskeleton of plants is introduced, which focuses on the structure and dynamics of MT, MF, and IF, and summarizes the crosslinking factors between MT and MF. Also, the paper prospects the future research direction of plant cytoskeleton and the possible research hotspot, which provides a certain reference for people to continue to explore the function of plant cytoskeleton in the future.展开更多
Background:Cantharidin (CTD),a natural toxin produced from Chinese blister beetles,has extensive anti-tumor activity.The present study investigated the effect of CTD on a human colon cancer cell line to elucidate pote...Background:Cantharidin (CTD),a natural toxin produced from Chinese blister beetles,has extensive anti-tumor activity.The present study investigated the effect of CTD on a human colon cancer cell line to elucidate potential new insights regarding the mechanism(s) through which CTD exerts its anti-tumor effects.Materials and methods:The inhibitory effect of CTD on human colon cancer HCT116 cells was evaluated using the IncuCyte ZOOMTM analyzer.Apoptotic cells were detected by Annexin V-FITC/PI assay and cell cycle was evaluated with flow cytometry following propidium iodide staining.Alterations in F-actin microfilaments were analyzed by FITC-phalloidin staining and morphological changes were evaluated with a laser scanning confocal microscope.Cell migration assay was carried out to investigate the effects of CTD on migration of HCT116 cells in vitro.Results:CTD exhibited a significant growth inhibitory effect on HCT116 cells accompanied by an increase in G2/M phase cells,without a significant effect on apoptosis.CTD-treated cells also exhibited a dramatic collapse in their microfilament network and a significant reduction in cell adhesion.Conclusion:CTD inhibits growth by increasing G2/M phase cells and decreasing S phase cells,revealing that CTD exerts a significant growth inhibitory effect primarily through an inhibition of cell cycle progression (a cytostatic effect).Moreover,a negative effect on cell migration may also constitute a contributing factor to its anti-tumor potential.These findings suggest the potential use for developing CTD as a novel anti-cancer therapy that targets metastasis Giving full play to CTD may inhibit tumor transfer.展开更多
基金funded by the Gerald Kerkut Charitable Trust (GKT)(to BR)
文摘Aging is the leading risk factor for Alzheimer’s disease and other neurodegenerative diseases. We now understand that a breakdown in the neuronal cytoskeleton, mainly underpinned by protein modifications leading to the destabilization of microtubules, is central to the pathogenesis of Alzheimer’s disease. This is accompanied by morphological defects across the somatodendritic compartment, axon, and synapse. However, knowledge of what occurs to the microtubule cytoskeleton and morphology of the neuron during physiological aging is comparatively poor. Several recent studies have suggested that there is an age-related increase in the phosphorylation of the key microtubule stabilizing protein tau, a modification, which is known to destabilize the cytoskeleton in Alzheimer’s disease. This indicates that the cytoskeleton and potentially other neuronal structures reliant on the cytoskeleton become functionally compromised during normal physiological aging. The current literature shows age-related reductions in synaptic spine density and shifts in synaptic spine conformation which might explain age-related synaptic functional deficits. However, knowledge of what occurs to the microtubular and actin cytoskeleton, with increasing age is extremely limited. When considering the somatodendritic compartment, a regression in dendrites and loss of dendritic length and volume is reported whilst a reduction in soma volume/size is often seen. However, research into cytoskeletal change is limited to a handful of studies demonstrating reductions in and mislocalizations of microtubule-associated proteins with just one study directly exploring the integrity of the microtubules. In the axon, an increase in axonal diameter and age-related appearance of swellings is reported but like the dendrites, just one study investigates the microtubules directly with others reporting loss or mislocalization of microtubule-associated proteins. Though these are the general trends reported, there are clear disparities between model organisms and brain regions that are worthy of further investigation. Additionally, longitudinal studies of neuronal/cytoskeletal aging should also investigate whether these age-related changes contribute not just to vulnerability to disease but also to the decline in nervous system function and behavioral output that all organisms experience. This will highlight the utility, if any, of cytoskeletal fortification for the promotion of healthy neuronal aging and potential protection against age-related neurodegenerative disease. This review seeks to summarize what is currently known about the physiological aging of the neuron and microtubular cytoskeleton in the hope of uncovering mechanisms underpinning age-related risk to disease.
文摘BACKGROUND Glioma is one of the most common intracranial tumors,characterized by invasive growth and poor prognosis.Actin cytoskeletal rearrangement is an essential event of tumor cell migration.The actin dynamics-related protein scinderin(SCIN)has been reported to be closely related to tumor cell migration and invasion in several cancers.AIM To investigate the role and mechanism of SCIN in glioma.METHODS The expression and clinical significance of SCIN in glioma were analyzed based on public databases.SCIN expression was examined using real-time quantitative polymerase chain reaction and Western blotting.Gene silencing was performed using short hairpin RNA transfection.Cell viability,migration,and invasion were assessed using cell counting kit 8 assay,wound healing,and Matrigel invasion assays,respectively.F-actin cytoskeleton organization was assessed using F-actin staining.RESULTS SCIN expression was significantly elevated in glioma,and high levels of SCIN were associated with advanced tumor grade and wild-type isocitrate dehydrogenase.Furthermore,SCIN-deficient cells exhibited decreased proliferation,migration,and invasion in U87 and U251 cells.Moreover,knockdown of SCIN inhibited the RhoA/focal adhesion kinase(FAK)signaling to promote F-actin depolymerization in U87 and U251 cells.CONCLUSION SCIN modulates the actin cytoskeleton via activating RhoA/FAK signaling,thereby promoting the migration and invasion of glioma cells.This study identified the cancer-promoting effect of SCIN and provided a potential therapeutic target for the treatment of glioma.
基金funded by"Beijing Natural Science Foundation,grant number 6224060","Young Elite Scientists Sponsorship Program by BAST,grant number",BYESS2023192"Program of Beijing Municipal Education Commission,grant number KM202310020006"+1 种基金"Bejing University of Agriculture science and Technology innovation Sparkling support plan,grant number,BUA-HHXD2022007""2022 Research and Innovation ability improvement plan for young teachers of Beijing University of Agriculture,grant number QJKC2022028".
文摘Anticancer drugs are one of the most direct means of cancer therapy.However,the various cancer progressions hamper the development and discovery of anticancer drugs.In fact,the mechanical properties of the tumor cytoskeleton are extremely vital for any phase of cancer,especially in tumor invasion and metastasis.However,in the current category of anticancer drugs,the cytoskeleton-targeting drugs are limited and their role in tumor progression is unclear.Here,we present the mechanical characteristics of tumor stiffness that are tightly regulated by the cancer cytoskeleton,including actin filaments and microtubules during tumor initiation,growth and metastasis,and review the natural drugs that target the cancer cytoskeleton.We define cytoskeleton dynamics as target mechanisms for anticancer drugs and summarize the plant,microbial and marine sources of natural products.Furthermore,this paper also provides a material pathway to study active tumor mechanics,and introduces the unique advantages and future application potential of tumor cytoskeleton-targeting drugs in clinical use.The material approaches to active cancer mechanics are supplied in this review.We aim to promote the development of anticancer drugs that target tumor mechanics by using those material approaches and finding their pharmacological application.
基金the National Natural Science Foundation of China(81473552 and 81774031)partial experiment was funded by the Beijing University of Chinese Medicine(2018-JYBZZ-XS011).
文摘Background:In myocardial ischemia,hypoxia leads to destruction of the cytoskeleton,and especially the imbalance of microtubule polymerization-depolymerization,which seriously affects the structure and function of cardiomyocytes.We previously showed that a Yiqi Huoxue Decoction (YQHX) improves mitochondrial function and decreases anti-oxidative effects in hypoxia-induced H9c2 cell injury.Therefore,in this study we investigated whether YQHX protects against hypoxia-induced damage by decreasing damage to the cardiac cytoskeleton.Methods:After reaching 70%-80% confluence,H9c2 cells were synchronized in serum-free Dulbecco's Modified Eagle Medium for 6 hours,then divided into control,model,and YQHX (100,200,400 μg/mL) groups,which were then grown in a hypoxic atmosphere for 12 hours.Cardiac cell viability was assessed using an xCELLigence system.The levels of lactate dehydrogenase,maleic dialdehyde,and superoxide dismutase in H9c2 cell supernatants were measured.Hoechst 33258 staining was employed to observe cardiac cell apoptosis.Confocal microscopy,immunofluorescence,and western blot analysis were performed to evaluate the protective effects of the YQHX against hypoxia-induced injury in the H9c2 cell line.Results:Cells that were pretreated with YQHX were more able to maintain their microtubule structure in the early stages of hypoxia and had better myocardial fitness in response to hypoxia compared with cells that were not pretreated.However,hypoxia-induced upregulation of α-tubulin and β-tubulin expression antagonized the protective effect of YQHX (100 μg/mL).In addition,YQHX (100 μg/mL) treatment significantly upregulated MAP4 protein expression (P =.003) and downregulated p-AMPKα protein expression (P <.001) compared with the model group.Conclusion:The results indicate that YQHX plays a role in protecting against oxidative stress injury and apoptosis in H9c2 cells.Notably,our results suggested that the YQHX could mitigate the damage to the cardiac cytoskeleton and the dysregulation of AMPK-related protein signaling pathways that are induced by hypoxia.
基金funded by the National Natural Science Foundation of China ( 31660258, 31771014,31860262,11762006)the Science and Technology Foundation of Guizhou Province ( 2019-2787,2018-1412, 2016-5676,2017-5718)+2 种基金the Science and Technology Innovative Talent Team of Guizhou Province ( 2015-4021)the 2011 Collaborative Innovation Program of Guizhou Province ( 2015-04 )the Cell and Gene Engineering Innovative Research Groups of Guizhou Province ( KY-2016-031)
文摘Objective Fumonisin B1(FB1)is an important mycotoxin in nature worldwide.The biomechanical properties of cells are closely related to their structure and function,and the cytoskeleton is the structural and functional basis of cells motility,and therefore,from a biomechanical point of view,the purpose of this study is to investigate the effects of FB1 on the biomechanical properties,migration capacity and cytoskeletal structure of human umbilical vein endothelial cells(HUVECs),which may lay an experimental foundation for further exploration of the toxicity mechanism of fumonisin.Methods HUVECs were cultured and treated with different concentrations of FB1.Then,CCK-8 kit was used to detect the effect of FB1 on the survival rate.The osmotic fragility of the cells was measured after treatment with different osmotic pressures for30 min.The cell membrane fluidity was measured by fluorescence polarization method.The cell electrophoretic mobility was measured by cell electrophoretic apparatus.The migration capacity of the cells was observed by scratch repair assay.The changes of reactive oxygen species and cytoskeletal structure were observed by confocal laser scanning microscopy.Finally,the mRNA and protein relative expression levels of cytoskeletal binding proteins were detected by real-time PCR,Western blotting and confocal laser scanning.Results The results of CCK-8 showed that FB1 could significantly inhibit the proliferation of HUVECs in a dose-and time-dependent manner.After treatment of HUVECs with FB1,the hypotonic resistance of the cell,cell surface charge,cell membrane fluidity and migration capacity were all weakened,while reactive oxygen species were significantly increased and the cytoskeletal structure was significantly reorganized.Furthermore,RTPCR results showed that the mRNA relative expression levels of cytoskeletal binding proteins,exception of actin,were down-regulated after treated with FB1.Besides,Western blotting and statistical analysis based on fluorescence intensity of laser confocal microscopy confirmed theses changes in protein level.Conclusions FB1 can significantly affect the biomechanical properties and motility of HUVECs,which may be directly correlated to the remodel of F-actin cytoskeleton,as well as the relative expression changes of cytoskeletal binding proteins.It is significant for further exploring the toxicity mechanism of fumonisin.
基金funded by the National Natural Science Foundation of China ( 31660258,31771014, 31860262,31570938,31260227)the Science and Technology Foundation of Guizhou Province ( 2019-2787,2018-1412, 2016-5676,2017-5718)+2 种基金the Science and Technology Innovative Talent Team of Guizhou Province ( 2015-4021)the 2011 Collaborative Innovation Program of Guizhou Province ( 2015-04 )the Cell and Gene Engineering Innovative Research Groups of Guizhou Province ( KY-2016-031)
文摘Background Dendritic cells(DCs)are the most important antigen-presenting cells due to their professional and extremely efficient antigen-presenting function.The dynamics of cytoskeleton plays crucial regulated roles on DCs’immune functions and biophysical properties.Several evidences show that tumor-derived suppressive cytokines deteriorate DCs’immune functions through remodeling their F-actin cytoskeleton.But the underlying mechanism is still elusive.Tropomodulin1(Tmod1),a cytoskeleton-binding protein,regulates and stabilizes actin filaments lengths and cytoskeleton architecture,which involves in the regulations of the morphology,formation of neural dendrites and biophysical properties of cells.Our previous studies found that mature DCs(mDCs)had a higher expression of Tmod1 than immature DCs(imDCs). Therefore,it’s hypothesized that Tmod1 maybe involve in the modification of DCs’functions.Objective The aim of the study is to investigate the effects of Tmodl on the immune functions and biophysical properties of DCs and the underlying mechanisms in order to further understand the biological behaviors of DCs.Methods Bone marrow-derived cells were harvested from wild type(C57BL/6 J)mice and Tmod1 knockout mice(Tmod1 overexpressing transgenic(TOT)/Tmod1-/-)and differentiated to immature dendritic cells(imDCs)by rmGM-CSF and rmIL-4.imDCs were then matured by lipopolysaccharides(LPS)treatment.The expressions of the surface markers in DCs,including CD80,CD86,CD40,MHC-Ⅱand CCR7,were detected by flow cytometry,Western blot and qRT-PCR.The inflammation cytokines such as IL-6,IFN-γ,IFN-βand IL-10 were also detected by flow cytometry.The immune functions and the biophysical properties of DCs were compared between the wild type and Tmod1 knockout mice.The F-actin content and dendritic pseudopodia of these two kinds of DCs were detected by flow cytometry and laser scanning confocal microscope respectively.Finally,we detected the MyD88 dependent and independent signaling pathway to discover the molecular mechanisms.Results We found that Tmod1-deficient mDCs showed deficient antigen-presenting ability and they failed to express enough MHC-Ⅱ,co-stimulated molecules(CD80/86,CD40)and CCR7 on their cell surface.The secretions of the inflammatory cytokines IL-6 and IFN-γwere decreased while the anti-inflammatory cytokines IFN-βand IL-10 were increased in the supernatant of Tmod1-deficient mDCs.As compared to DCs of wild type mice,the migration ability of DCs from Tmod1 knockout mice were dramatically damaged including their free migration and CCL19 mediated chemotaxis migration.However,we found that Tmod1 knockout had no effects on the imDCs’endocytosis ability.Furthermore,Tmod1 knockout DCs showed higher osmotic fragility,lower Young’s modulus,less F-actin content and shorter dendritic pseudopodia.Under LPS stimulation,the phosphorylation level of p65 and p38 were significantly downregulated in Tmod1 knockout mice while the expression of p-IRF3 was upregulated.Conclusions These results indicated that Tmodl knockout leads to deficient antigen-presenting ability and impaired migration of DCs as well as their biophysical properties.The underlying mechanisms are due to the inhibitions of the TLR4-mediated NF-κB and p38 MAPK singling pathway and the activation of the IRF3 signaling pathway,as well as the disturbed reorganization of the F-actin cytoskeleton.Our results provide a new insight on the functions of Tmod1 which can affect the DCs’immune functions and biophysical properties through regulating the TLR4-mediated singling pathways and cytoskeleton remodeling.
基金supported by Department of Biotechnology,Department of Science and Technology,Government of India
文摘Objective:To examine the role of heat shock protein 90(Hsp90) in the maintenance of actin cytoskeleton in human neuroblastoma tumor cells.Methods:Co-precipitation experiments were performed to examine Hsp90 interaction with actin.Hsp90 and actin interactions were evaluated by protein refolding and acto-myosin motility assays.17-(AUylamino)-17- demethoxygeldanamycin(17AAG) induced actin-cytoskeleton re-organization was examined by laser scanning confocal microcopy.Results:It was shown that inhibition of Hsp90 by 17AAC accelerates detergent induced cell lysis of neuroblastoma tumor cells through destabilization of actin cytoskeleton.The in vitro co-precipitation experiments showed that functional but not mutant Hsp90 binds with F-actin.Among biochemical modifications,phopshorylation and oligomerization enhanced Hsp90 binding with F-actin.F-actin binding to Hsp90 interfered with Hsp90 chaperone activity in protein refolding assays,and Hsp90 binding to F-actin interfered with actin motility on myosin coated flow cell.In the combination treatment,17AAG irreversibly augmented the effect of cytochalasin D,an inhibitor of actin polymerization.Conclusions:It can be concluded that Hsp90 binds to F-actin in tumor cells and maintains the cellular integrity. The results display a novel element of Hsp90 inhibition in destabilizing the actin cytoskeleton of tumor cells,therefore suggest that 17AAG combination with cytoskeletal disruptor may be effective in combating cancer.
基金Supported by the Natural Science Foundation of China (No.81300768 No.81371048+4 种基金 No.81670853)Science and Technology Department of Sichuan Province of China (No.2015HH0031)Health and Family Planning Commission of Sichuan Province of China (No.100539 No. 090505 No.090514)
文摘AIM:To investigate the effects of dexamethasone(DEX) and 1-(5-isoquinolinesulfonyl)-homopiperazine(HA1077) on actin cytoskeleton and β-catenin in cultured human trabecular meshwork(HTM) cells.METHODS: The HTM cells were separated from human eyeball and cultured in vitro.They were divided into control group,DEX(1×10^(-6)mol/L) group,HA1077(3×10^(-5)mol/L)group,and DEX(1×10^(-6)mol/L) and HA1077(3×10^(-5)mol/L)group.Actin cytoskeleton and β-catenin in HTM cells of the four groups were examined by immunofluorescence and Western blot analyses.RESULTS: In DEX group,there were reorganization of actin cytoskeleton and formation of cross linked actin networks(CLANs),which were partially reversed in DEX and HA1077 group.DEX treatment also induced an increased expression of β-catenin,which was obviously reduced in DEX and HA1077 group.Meanwhile,the cultured HTM cells in HA1077 group had lower expression of β-catenin than that in the control group. CONCLUSION: Our results show that HA1077 can reverse the changes of actin organization and expression of β-catenin induced by DEX in cultured HTM cells,suggesting that HA1077 may play an important role in increasing outflow and reducing intraocular pressure.
文摘We have demonstrated that the distribution of microtubules (MT), mlcrofilaments (MF) and fibronectin (FN) were diminished, while the gene expression of the calmodulin and c- fos enhanced in the transformed C3H10T1/2 cells. After treatment with 1 mM db-cAMP for 1 hour and 2 hours, there was an early and repldly reduced in gene expression of Calmodulin and c-fos respectively. After db-cAMP treatment for 4 -5 days, the number of capping cells of ConA binding decreased significantly and the cell surface microvllll decreased as well. The growth of treated cells was inhibited markedly. By using 4F1 cDNA probe, which is preferentially expressed In G1 phase, we have found that the db- cAMP treated cells were accumulated at G1 phase. Of particular interest is the fact that the distribution of microtubules, mlcrofilaments and fibronectln were recovered after treatment with 1 mM db-cAMP for 6 days. It is suggested that the Inhibition of proliferation, alteration, of phenotype and reco- very of cytoskeleton is
文摘To evaluate the effects of Hantaan(HT) virus on the cytoskeleton of human endothelial cells(HECs).The human umbilical vein endothelial cells were isolated and cultured in vitro.After confluence,the SR-ll strain of HT virus(100 and,300 TCID_(50)) were inoc
基金the Innovative Research Team of Taizhou Polytechnic College(No.TZYTD-16-4)Natural Science Research General Project of Jiangsu Higher Education Institutions(No.18KJD350002)the Doctoral Research Foundation of Taizhou Polytechnic College(No.1322819004).
文摘Focal adhesions are polyproteins linked to extracellular matrix and cytoskeleton,which play an important role in the process of transforming force signals into intracellular chemical signals and subsequently triggering related physiological or pathological reactions.The cytoskeleton is a network of protein fibers in the cytoplasm,which is composed of microfilaments,microtubules,intermediate filaments,and cross-linked proteins.It is a very important structure for cells to maintain their basic morphology.This review summarizes the process of fluid shear stress transduction mediated by focal adhesion and the key role of the cytoskeleton in this process,which focuses on the focal adhesion and cytoskeleton systems.The important proteins involved in signal transduction in focal adhesion are introduced emphatically.The relationship between focal adhesion and mechanical transduction pathways are discussed.In this review,we discuss the relationship between fluid shear stress and associated diseases such as atherosclerosis,as well as its role in clinical research and drug development.
基金National Nature Science Foundation of China, No. 39290700-03P
文摘Objective: To study the mechanisms of the injury of the cytomembrane and cytoskeleton of cultured myocardial cells exposed to the combined stimulation of hypoxia and burn serum. Methods: The myocardial cells of neonatal rats were cultured in vitro. The changes of the cytoskeleton and cytomembrane of the cultured myocardial cells before and after the combined stimulation of hypoxia and burn serum were observed dynamically with immunohistochemistry, flow cytometry and viscoelasticity determination technique. The deformity of the cultured myocardial cells was determined mathematically. Results: In the early stage after the combined impact of hypoxia and burn serum, the cultured myocardial cells were deformed. The elastic coefficient k1 and k2 and the viscosity coefficient μ of the cytomembrane were significantly decreased (P < 0.05 - 0. 01 ). The fluorescent intensity of the cytoskeleton protein and the number of the intermediate filaments and imcrotubules were markedly decreased. Conclusion: The cytoskeleton is essential to the integrity of the cytomembrane. The damage of the cytoskeleton induces the increase of fragility and decrease of viscoelasticity of the cytomembrane . These changes of the biomechanic characteistics participate directly in the initiation and development myocardial damage.
文摘The manuscript deals with our studies and experiences in the assessment of cytoskeleton in different cellular models and situations.The immunofluorescent study of several cytoskeletal proteins was relevant in the evaluation of a therapy for osteoarthritis,in case of alkaptonuria and in testing the efficacy of docetaxel in neuroblastoma cancer cells leading to apoptosis.A relevant part of our experience focus on the study of cytoskeleton in seminiferous epithelium and spermatozoa,identifying alterations affecting blood-testis barrier after a silver nanoparticle treatment,chromosomal segregation in case of varicocele,sperm motility and diagnosing systematic sperm defects as“Primary ciliary dyskinesia”and“Dysplasia of the fibrous sheath”.The evaluation of cytoskeleton represents a specific and sensitive analysis in establishing the health status of different cells.
基金This research was supported by the School of Science and Technology(CCyT-2021-5)from the Autonomous University of Mexico City(UACM).
文摘Cytoskeletal remodeling affects the shape,adhesion,and motility of cells.Cytoskeletal dynamics are modulated by matrix proteins,integrins,and several cytokines in the tumor microenvironment.In this scenario,signaling is activated by integrins and interferons,which can induce ISG15 gene expression.This gene encodes a ubiquitin-like protein that functions as a protein modifier via the ISGylation system.Furthermore,non-conjugated ISG15 acts as a cytokine-like protein.In this viewpoint,the interplay between free ISG15,protein ISGylation,and cytoskeletal dynamics in the tumor microenvironment is discussed.
文摘Two different simple cases of plane tensegrity cytoskeleton geometries are presented and investigated in terms of stability. The tensegrity frames are used to model adherent cell cytoskeletal behaviour under the application of plane substrate stretching and describe thoroughly the experimentally observed reorientation phenomenon. Both models comprise two elastic bars (microtubules), four elastic strings (actin filaments) and are attached on an elastic substrate. In the absence of external loading shape stability of the cytoskeleton is dominated by its prestress. Upon application of external loading, the cytoskeleton is reorganized in a new direction such that its total potential energy is rendered a global minimum. Considering linear constitutive relations, yet large deformations, it is revealed that the reorientation phenomenon can be successfully treated as a problem of ma- thematical stability. It is found that apart from the magnitude of contractile prestress and the magnitude of extracellular stretching, the reorientation is strongly shape–dependent as well. Numerical applications not only justify laboratory data reported in literature but such experimental evidence as the concurrent appearance of two distinct and symmetric directions of orientation, indicating the cellular coexistence of phases phenomenon, are clearly detected and incorporated in the proposed mathematical treatment.
基金Guangxi Natural Science Foundation Project(No.2020GXNSFAA238012)2021"Qihuang Project"High-level Talent Team Cultivation Project(2021006)2021 Graduate Education Innovation Project(No.YCXJ2021047)。
文摘Objective:To observe the effect of lipopolysaccharide(LPS)on cytoskeleton and the effect of Dahuang Lingxianfang on nF-KB/MAPK signaling pathway.Methods:Biliary epithelial cells of each group were stained with photolipin fluorescent staining,and the arrangement of cytoskeleton was observed under laser confocal microscope.Western blotting was used to detect the expression of f-actin.Results:After LPS intervention,the biliary epithelial cells showed nuclear shrinkage or damage,and the skeleton was broken or lumped.The cytoskeleton was partially repaired after the intervention of pathway blocking preparation combined with RHUbarb Lingxianfang.All the other groups had different degree of cytoskeleton fracture.Compared with normal group,the expression of F-actin protein in LPS group was decreased(P<0.05);Compared with LPS group,the expression of F-actin in LPS+TCM group,LPS+PDTC+TCM group,LPS+SB203580+TCM group and LPS+PDTC+SB203580+TCM group was significantly increased(P<0.05);Compared with traditional Chinese medicine group,the expression of F-actin in LPS+PDTC+traditional Chinese medicine group,LPS+SB203580+traditional Chinese medicine group and LPS+PDTC+SB203580+traditional Chinese medicine group had no significant difference(P>0.05).Conclusion:RHUbarb Lingxianfang can restore the sequence of biliary epithelial cytoskeleton and protect its microfilament structure under inflammation,and its mechanism may be related to the regulation of NF-KB/MAPK signaling pathway.
文摘Cytoskeleton exists in all eukaryotes and is involved in many significant cytobiological processes, especially the movements and developmental changes of plant cells. The cytoskeleton consists of microtubule (MT), microfilament (MF), and intermediate filament (IF). MT and MF are vital components of plant cytoskeleton. Crosslinking factor acts as a bridge between MF and MT. They play an important role in cellular life process and have always been a hot topic and key point in plant cytobiology, and the IF is a difficult point in this field. In this paper, the latest research on the cytoskeleton of plants is introduced, which focuses on the structure and dynamics of MT, MF, and IF, and summarizes the crosslinking factors between MT and MF. Also, the paper prospects the future research direction of plant cytoskeleton and the possible research hotspot, which provides a certain reference for people to continue to explore the function of plant cytoskeleton in the future.
文摘Background:Cantharidin (CTD),a natural toxin produced from Chinese blister beetles,has extensive anti-tumor activity.The present study investigated the effect of CTD on a human colon cancer cell line to elucidate potential new insights regarding the mechanism(s) through which CTD exerts its anti-tumor effects.Materials and methods:The inhibitory effect of CTD on human colon cancer HCT116 cells was evaluated using the IncuCyte ZOOMTM analyzer.Apoptotic cells were detected by Annexin V-FITC/PI assay and cell cycle was evaluated with flow cytometry following propidium iodide staining.Alterations in F-actin microfilaments were analyzed by FITC-phalloidin staining and morphological changes were evaluated with a laser scanning confocal microscope.Cell migration assay was carried out to investigate the effects of CTD on migration of HCT116 cells in vitro.Results:CTD exhibited a significant growth inhibitory effect on HCT116 cells accompanied by an increase in G2/M phase cells,without a significant effect on apoptosis.CTD-treated cells also exhibited a dramatic collapse in their microfilament network and a significant reduction in cell adhesion.Conclusion:CTD inhibits growth by increasing G2/M phase cells and decreasing S phase cells,revealing that CTD exerts a significant growth inhibitory effect primarily through an inhibition of cell cycle progression (a cytostatic effect).Moreover,a negative effect on cell migration may also constitute a contributing factor to its anti-tumor potential.These findings suggest the potential use for developing CTD as a novel anti-cancer therapy that targets metastasis Giving full play to CTD may inhibit tumor transfer.
