Analysis and Review of Downregulated Actin Cytoskeletal Proteins in Non-Small Cell Lung Cancer

Actin, a highly conserved protein, plays a dominant role in Non-small cell lung cancer (NSCLC). Late diagnosis and the aggressive nature of NSCLC pose a significant threat. Studying the clinic pathological properties of NSCLC proteins is a potential alternative for developing treatment strategies. Towards this, 35 downregulated actin cytoskeletal proteins on NSCLC prognosis and treatment were studied by examining their protein-protein interactions, gene ontology enrichment terms, and signaling pathways. Using PubMed, various proteins in NSCLC were identified. The protein-protein interactions and functional associations of these proteins were examined using the STRING database. The focal adhesion signaling pathway was selected from all available KEGG and Wiki pathways because of its role in regulating gene expression, facilitating cell movement and reproduction, and significantly impacting NSCLC. The protein-protein interaction network of the 35 downregulated actin cytoskeleton proteins revealed that ACTG1, ACTR2, ACTR3, ANXA2, ARPC4, FLNA, TLN1, CALD1, MYL6, MYH9, MYH10, TPM1, TPM3, TPM4, PFN1, IQGAP1, MSN, and ZXY exhibited the highest number of interactions. Whereas HSPB1, CTNNA1, KRT17, KRT7, FLNB, SEPT2, and TUBA1B displayed medium interactions, while UTRN, TUBA1B, and DUSP23 had relatively fewer interactions. It was discovered that focal adhesions are critical in connecting membrane receptors with the actin cytoskeleton. In addition, protein kinases, phosphatases, and adapter proteins were identified as key signaling molecules in this process, greatly influencing cell shape, motility, and gene expression. Our analysis shows that the focal adhesion pathway plays a crucial role in NSCLC and is essential for developing effective treatment strategies and improving patient outcomes.

Effect of glycosides of Cistanche on the expression of mitochondrial precursor protein and keratin type Ⅱ cytoskeletal 6A in a rat model of vascular dementia

Glycosides of Cistanche（GC）is a preparation used extensively for its neuroprotective effect against neurological diseases,but its mechanisms of action remains incompletely understood.Here,we established a bilateral common carotid artery occlusion model of vascular dementia in rats and injected the model rats with a suspension of GC（10 mg/kg/day,intraperitoneally）for 14 consecutive days.Immunohistochemistry showed that GC significantly reduced p-tau and amyloid beta（Aβ）immunoreactivity in the hippocampus of the model rats.Proteomic analysis demonstrated upregulation of mitochondrial precursor protein and downregulation of keratin type II cytoskeletal6A after GC treatment compared with model rats that had received saline.Western blot assay confirmed these findings.Our results suggest that the neuroprotective effect of GC in vascular dementia occurs via the promotion of neuronal cytoskeleton regeneration.

Acrylamide Alters Cytoskeletal Protein Level in Rat Serum

To study biomarker of acrylamide （ACR） induced neuropathy, Wistar rats received 20 or 40 mg/kg of ACR by ip injection and the levels of light neurofilament （NF-L）, middle NF （NF-M）, heavy NF （NF-H）, β-actin, α-tubulin, and β-tubulin proteins in serum were evaluated using both SDS-PAGE and Western blotting. Compared to controls, NF-L and NF-M decreased,

Distinct Cytoskeletal Injuries Induced by As,Cd,Co,Cr,and Ni Compounds

The risks of metal compounds to human health are highlighted by the ubiquity of exposure and their persistence in the environment.Although compounds of As,Cd,Co,Cr,and Ni are known or“reasonably anticipated”to be carcinogenic to humans and/or experimental animals, the cellular targets of these health hazards and the underlying mechanisms of their carcinogenic- ity are still unclear.We show in this report that dramatic,time-and dose-dependent cytoskeletal perturbations,especially in the distribution and organization patterns of microtubules and mi- crofilaments,two of the principal components of the cytoskeleton,occurred in 3T3 cells upon exposure to these metal salts.Each metal salt appeared to induce a different,typical pattern of cytoskeletal injury,probably reflecting the specificity of action of each metal ion.These results suggest that the cytoskeleton can indeed act as a target for injury by epigenetic carcinogenic metal compounds in the environment.These findings should help our efforts to understand the mechanisms of action of metal compounds at the subcellular and molecular levels.1989 Academic Press,Inc.

Gene expression of 49 kDa apyrase,cytoskeletal proteins,ATPase,ADPase and amino acid contents of Pisum sativum(L.)cells germinated in Euryops arabicus(Steud.ex Jaub.&Spach)water extract

The present research reports of quick and marked changes induced by plant extract of Euryops arabicus in the gene expression of 49-kDa apyrases,cytoskeletal proteins,ATPases,ADPase and amount of amino acid of pea(Pisum sativum L.var.Alaska).Pellets of cytoskeletals proteins(27000 xg)were probed with anti-apyrase antibody,biotinylated anti-rat,actin and alpha and beta-tubulin for Western blotting.ATPase and ADPase activities were determined based on the hydrolytic efficacy of adenine triphosphate and adenine diphosphate.By 72 hours,the abundance of apyrases,cytoskeletal proteins and amount of amino acid in pellets of 27000 xg of germinated pea seeds in E.arabicus extracts were sharply increased than those sown in distilled water.All the samples exhibited that the stems had more amount from apyrases,cytoskeletal proteins,amino acids and ATPase and ADPase activities than primary leaves and primary roots that were germinated either on E.arabicus water extract or in distilled water.Based on the enzyme’s capability to hydrolyse nucleotide triphosphate and nucleotide diphosphate as well as the direct association between expression of 49-kDa apyrase and cytoskeletal proteins,E.arabicus water extract had an important effect on plant germinations.

Thermal Fluctuation Induced Piezoelectric Effect in Cytoskeletal Microtubules: Model for Energy Harvesting and Their Intracellular Communication

Cytoskeletal microtubules have long been conjectured to have piezoelectric properties. They have been shown to behave as nematic liquid crystals which oscillate along their director axis due to the prevalent thermal fluctuations. In this work, we develop a theoretical model of the mechanics of microtubules in the cytosolic space based on the buckling of its structure due to these thermal fluctuations. This cytosolic space has been considered as a viscoelastic medium in which microtubule oscillations have been considered. As a result of resilience of cytosol and neighbouring filaments from the axial force due to thermal fluctuations, the surface traction acting laterally on the microtubule structure has been further used to elucidate its piezoelectric behaviour in vivo. After the piezoelectric properties induced by thermal fluctuations (in addition to the buckling) of microtubules have been discussed, we propose a model discussing how microtubules behave as energy harvesters and communicate via electromagnetic radiation, with each other, in an intracellular environment.

IgA1 from HSP Patients Trigger Apoptosis and Inhibit Cytoskeletal Proteins in HUVEC

Background: Henoch-Schonlein purpura (HSP) is a kind of systemic small vessel vasculitis in children. Endothelium cells injury induced by IgA1 is considered important in the pathogenesis of HSP. Research found that the apoptosis of vein endothelial cells was related to the vasculitis in HSP patients. Purpose: To observe the effect of IgA1 from HSP patients on the apoptosis of HUVEC and firstly analyze the mechanism of the apoptosis of HUVEC induced by IgA1. Methods: HUVECs were cultured in 3 different conditional media with IgA1 from HSP patients, normal healthy children and simply medium (blank control). Serum IgA1 was purified by jacalin affinity chromatography. The rates of apoptosis in HUVEC incubated with IgA1 were determined by TUNEL method and flow cytometry, respectively. The expression of the cytoskeletal proteins, such as FAK, Vinculin and MLCK was detected with the methods of Real-time PCR and Westernblot, respectively. Results: The present study showed that the apoptosis rate of HUVEC by IgA1 isolated from HSP patients was higher than blank control (14.77% ± 2.23% vs 2.25% ± 0.77%) (P < 0.01) and the rate of HUVEC by IgA1 from normal healthy children was higher than blank control (9.97% ± 1.48% vs 2.25% ± 0.77%) (P < 0.01). The cytoskeletal proteins, such as FAK, Vinculin and MLCK expression were down-regulated in HUVEC co-cultured with IgA1 isolated from HSP patients for 24h. Conclusion: These findings firstly on IgA1 from HSP patients may induce apoptosis of vascular endothelial cells through inhibiting the cytoskeletal proteins expression. IgA1 may accelerate progression of HSP by inducing apoptosis of vascular endothelial cells.

Use of artificial neural networks to identify and analyze polymerized actin-based cytoskeletal structures in 3D confocal images

Background:Living cells need to undergo subtle shape adaptations in response to the topography of their substrates.These shape changes are mainly determined by reorganization of their internal cytoskeleton,with a major contribution from filamentous(F)actin.Bundles of F-actin play a major role in determining cell shape and their interaction with substrates,either as“stress fibers,”or as our newly discovered“Concave Actin Bundles”(CABs),which mainly occur while endothelial cells wrap micro-fibers in culture.Methods:To better understand the morphology and functions of these CABs,it is necessary to recognize and analyze as many of them as possible in complex cellular ensembles,which is a demanding and time-consuming task.In this study,we present a novel algorithm to automatically recognize CABs without further human intervention.We developed and employed a multilayer perceptron artificial neural network(“the recognizer”),which was trained to identify CABs.Results:The recognizer demonstrated high overall recognition rate and reliability in both randomized training,and in subsequent testing experiments.Conclusion:It would be an effective replacement for validation by visual detection which is both tedious and inherently prone to errors.

Towards a better recording of microtubule cytoskeletal spatial organization and dynamics in plant cells

Numerous fluorescent marker lines are currently available to visualize microtubule(MT)architecture and dynamics in living plant cells, such as markers expressing p35S::GFP-MBD or p35S::GFP-TUB6.However, these MT marker lines display obvious defects that affect plant growth or produce unstable fluorescent signals. Here, a series of new marker lines were developed, including the pTUB6::VisGreen-TUB6-expressing line in which TUB6 is under the control of its endogenous regulatory elements and e GFP is replaced with VisGreen, a brighter fluorescent protein. Moreover, two different markers were combined into one expression vector and developed two dual-marker lines.These marker lines produce bright, stable fluorescent signals in various tissues, and greatly shorten the screening process for generating dual-marker lines.These new marker lines provide a novel resource for MT research.

The p21-activated kinases in neural cytoskeletal remodeling and related neurological disorders

The serine/threonine p21-activated kinases(PAKs),as main effectors of the Rho GTPases Cdc42 and Rac,represent a group of important molecular switches linking the complex cytoskeletal networks to broad neural activity.PAKs show wide expression in the brain,but they differ in specific cell types,brain regions,and developmental stages.PAKs play an essential and differential role in controlling neural cytoskeletal remodeling and are related to the development and fate of neurons as well as the structural and functional plasticity of dendritic spines.PAK-mediated actin signaling and interacting functional networks represent a common pathway frequently affected in multiple neurodevelopmental and neurodegenerative disorders.Considering specific small-molecule agonists and inhibitors for PAKs have been developed in cancer treatment,comprehensive knowledge about the role of PAKs in neural cytoskeletal remodeling will promote our understanding of the complex mechanisms underlying neurological diseases,which may also represent potential therapeutic targets of these diseases.

Reorganization of cytoskeletal proteins of mouse oocytes mediated by integrins

To study whether integrins on cell membrane ligate with intracellular cytoskeletal proteins and mediate their reorganization in egg activation, female mice were used for su- perovulation. The zona-free oocytes were incubated separately with specific ligand of integrins, an active RGD peptide, in vitro for certain period of time. RGE peptide and mouse capacitated sperm were used as controls. Freshly ovulated oocytes and those treated with different factors were immunostained with FITC-labeled anti-actin antibody, then detected with confocal micro- scope. The results demonstrated that freshly ovulated mouse oocytes, oocytes incubated for 2 h in vitro and those treated with control RGE peptide for 15 min showed hardly visible fluorescene or only thin fluorescence in plasma membrane region. Oocytes coincubated with sperms for 15 min and those treated with active RGD peptide for 10 min, 30 min and 2 hours respectively had strong and thick fluorescence in the plasma membrane and cortical region of oocytes, and some of them showed asymmetrically fluorescent distribution. It is proved that integrins on membrane are ligated directly with cytoskeletal protein. Integrins binding with their ligands regulate reor- ganization of cytoskelal protein, which may be involved in transmembrane signaling in egg acti- vation.

Synthesized flavanoid-derived ligand reduced dengue virus type-2 replication in vitro

Objective: To investigate the antiviral property of a lead ligand, YK51 that was synthesized based on the flavanoid of a natural product toward dengue virus type-2(DENV2)replication.Methods: c RNA was isolated from HepG2 cells inoculated with 1 000 median tissue culture infective dose of DENV2 and treated with different doses of the ligand followed by RT-PCR to quantify the virus gene copies. Confocal microscopy of actin and tubulin redistribution was also performed.Results: The quantitative RT-PCR result showed reduction of the DENV2 gene copies as the ligand concentration was increased. The confocal microscopy result showed increase in the tubulin intensity(79.6%) of infected BHK21 cells treated with the ligand,compared with the non-treated cells(54.8%). The 1.5-fold increase in the intensity of tubulin suggested that the ligand inhibitory effect stabilized the cellular microtubule structure.Conclusions: The synthesized ligand YK51 reduced DENV2 viral load by inhibiting virus replication thus is highly potential to be developed as antiviral agent.

Impact of chitosan-based nanocarriers on cytoskeleton dynamics:Current status and challenges

Chitosan-based nanocarriers(CS-NCs)show a promising role in improving drugs and bioactive compounds delivery for therapy.However,the effects exerted by CS-NCs at the cellular level,including their recognition and uptake,have not been fully investigated yet.Many factors,including size,shape,concentration,and surface chemistry of CS-NCs,play an important role in determining the types of intracellular signals triggered.The mechanism of uptake and the involvement of the cytoskeleton during the CS-NCs endocytosis variates among the different cell types as well as further effects observed inside cells.In the present work,we discuss the effects induced by CS-NCs per se on the cytoskeleton,a key component in cell architecture and physiology.The focus of this report is made on tumoral and normal biological models in which CS-NCs could differentially affect the cell cytoskeleton.The recent years reports regarding the impact of CS-NCs on cytoskeleton dynamics and the current techniques for its evaluation are summarized and discussed.Understanding mechanisms underlying cytoskeletal impact after cell exposure to CS-NCs is critical for the design of safest value-added formulations in the biomedical field.Furthermore,this revision points out some interesting aspects of cytoskeletal changes and cell death encompassing anti-tumoral effects.

Protective Effect of Space Ginseng Yeast on Human Primary Fibroblasts under Simulated Microgravity Condition

[Objectives] To study the protective effect of Space Ginseng Yeast on human primary fibroblasts. [Methods] Multiple indicators were used to simulate the microgravity effect,so as to study the protective effect of Space Ginseng Yeast on human primary fibroblasts under simulated microgravity condition. [Results]Space Ginseng Yeast could improve changes of cell morphology and decline of the survival rate caused by microgravity,restore the expression and effective arrangement of cytoskeletal protein Tubulin,extracellular matrix proteins Integrin and Fibronectin,and can inhibit the abnormal expression of MMP-1. The protective effect of Space Ginseng Yeast diluted 20-500 times is better,and the protective effect of Space Ginseng Yeast diluted 100 times is the most obvious. [Conclusions] Space Ginseng Yeast can protect damage of human primary fibroblasts caused by simulated microgravity.

Glucocorticoid-induced alterations in titin, nebulin, myosin heavy chain isoform content and viscoelastic properties of rat skeletal muscle

Viscoelastic properties of skeletal muscle are associated with a complex network of cytoskeletal proteins where titin and nebulin play a substantial role. The need for evaluation of muscle viscoelastic properties is widely accepted in clinical use to evaluate the effect of treatment or progression of muscle pathology (atrophy). We tested the hypothesis that the viscoelastic properties (elasticity, tone and stiffness) change in atrophied muscles with concomitant changes in cytoskeletal proteins (titin, nebulin) and contractile protein (myosin heavy chain) proportion. Sixteen 24- week-old male rats of the Wistar strain were randomly allocated to two groups: dexamethasone group treated each day for 10 consecutive days with dexamethasone in order to induce atrophy and control group. Skeletal muscle viscoelastic properties (elasticity, tone and stiffness) were determined using a myotonometer. Titin, nebulin and myosin heavy chain content were quantified using SDS-PAGE electrophoresis. We found that glucocorticoid-induced muscle atrophy is accompanied by reduced elasticity and increased tone and stiffness, with concomitant changes in titin, nebulin and myosin heavy chain con- tent. The elasticity decreased by 10.9% (P P < 0.05), and stiffness was significantly lower in dexamethasone group (627.3 N/m vs 758.6 N/m);(P < 0.05). Compared with the control group, the content of titin, nebulin and myosin heavy chain in atrophied muscle was 76.4%, 70.6% and 82.3%, respectively. Our results may lead to a better understanding of the mechanism of muscle atrophy and provide better guidance for rehabilitation practices and help to find rational therapeutic intervention in the future.

MicroRNA-142-3p and microRNA-142-5p are downregulated in hepatocellular carcinoma and exhibit synergistic effects on cell motility

MicroRNAs （miRNAs）, an important class of small non-coding RNAs, regulate gene expression at the post-transcriptional level, miRNAs are involved in a wide range of biological processes and implicated in different diseases, including cancers. In this study, miRNA profiling and qRT-PCR validation revealed that miR-142-3p and miR-142-5p were significantly downregulated in hepatocellular carcinoma （HCC） and their expression levels decreased as the disease progressed. The ectopic expression of miR-142 significantly reduced HCC cell migration and invasion. Overexpression of either miR-142-3p or miR-142-5p suppressed HCC cell migration, and overexpression of both synergistically inhibited cell migration, which indicated that miR-142-3p and miR-142-5p may cooperatively regulate cell movement, miR-142-3p and miR-142-5p, which are mature miRNAs derived from the 3＇- and 5＇-strands of the precursor miR-142, target distinct pools of genes because of their different seed sequences. Pathway enrichment analysis showed a strong association of the putative gene targets of miR-142-3p and miR-142-5p with several cell motility-associated pathways, including those regulating actin cytoskeleton, adherens junctions, and focal adhesion. Importantly, a number of the putative gene targets were also significantly upregulated in human HCC cells. Moreover, overexpression of miR-142 significantly abrogated stress fiber formation in HCC cells and led to cell shrinkage. This study shows that mature miR-142 pairs collaboratively regulate different components of distinct signaling cascades and therefore affects the motility of HCC cells.

Effect of manipulation on cartilage in rats with knee osteoarthritis based on the Rho-associated protein kinase/LIM kinase 1/Cofilin signaling pathways

OBJECTIVE:To investigate the effect of manipulation treatment on knee osteoarthritis rats and the effect on Rho-associated protein kinase(ROCK)/LIM-kinase1(LIMK1)/Cofilin signaling pathway.METHOD:Fifty Specific pathogen Free Sprague-Dawley rats were randomly divided into five groups(n=8 each):blank group,model group,manipulation group,celecoxib group,and manipulation combined with celecoxib group(MC group).The osteoarthritis model was established by injecting 0.2 m L 4%papain into the articular disc of the rats.After successfully establishing the model,we treated the manipulation group with pushing manipulation using one-finger-meditation to the Neixiyan(EX-LE4),Waixiyan(EX-LE5),Xuehai(SP10),Liangqiu(ST34),and Zusanli(ST36)acupoints for 10 min each time.Also,the celecoxib group was gavaged with 24 mg·kg^(-1)·d^(-1 )celecoxib,while the MC group was treated using both of these two methods.After four weeks,the cartilage of the right femur was removed for hematoxylin-eosin staining of the cartilage tissue.The expressions of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in serum were observed using the enzyme-linked immunosorbent assay.Besides,we detected the expressions of ROCK,LIMK1,Phospho-LIM-kinase1(Phospho-LIMK1),Cofilin,and Phospho-Cofilin by Western blot.RESULTS:Compared to the model group,the manipulation group,celecoxib group,and MC group all exhibited superior results concerning pathological morphologic changes of cartilage,as observed by hematoxylin-eosin staining and calculated using the Mankin score.Besides,in contrast to the blank group,the model group exhibited elevated serum levels of IL-1βand TNF-α(P<0.01),while the expression of ROCK,LIMK1,Phospho-LIMK1,Cofilin,and Phospho-Cofilin in cartilage were all higher(P<0.01).Also,the serum levels of IL-1βand TNF-αin each treatment group were lower(P<0.01)than in the model group.Moreover,there were lower expressions of ROCK,LIMK1,Phospho-LIMK1,Cofilin,and Phospho-Cofilin in cartilage in the manipulation group and the MC group(P<0.01).Compared with the model group,the expression of ROCK,LIMK1,PhosphoLIMK1,Cofilin,and Phospho-Cofilin in cartilage in the celecoxib group were not statistically different(P>0.05).CONCLUSION:In this study,we established that manipulation has a better curative effect than celecoxib.Manipulation inhibits the development of cytoskeleton damage in cartilage and slows articular degeneration by regulating the expression of related proteins in the cytoskeletal signaling pathway.

Joining actions: crosstalk between intermediate filaments and actin orchestrates cellular physical dynamics and signaling

Many key cellular functions are regulated by the interplay of three distinct cytoskeletal networks, made of actin filaments,microtubules, and intermediate filaments(IFs), which is a hitherto poorly investigated area of research. However, there are growing evidence in the last few years showing that the IFs cooperate with actin filaments to exhibit strongly coupled functions.This review recapitulates our current knowledge on how the crosstalk between IFs and actin filaments modulates the migration properties, mechano-responsiveness and signaling transduction of cells, from both biophysical and biochemical point of view.

Effect of Electroacupuncture on Spermatogenesis in Rats with Oligozoospermia of Insufficiency of Shen(Kidney) Essence Syndrome

Objective: To assess the effect of electroacupuncture(EA) on expression of cytoskeletal proteins from Sertoli cells(SCs) and spermatogenesis in rats with oligozoospermia of insufficiency of Shen(Kidney)essence syndrome(OIKES).Methods: Twenty healthy male Sprague-Dawley rats were randomly assigned to four groups using a random number table: control,tripterygium glycosides(TG) treatment,sham and EA groups(n=5 in each group).A rat model of OIKES was established by oral gavage with TG.The EA group was treated with TG and received EA at Shenshu(BL 23) and Zusanli(ST 36) acupoints for 20 min,once daily for 30 days,while the sham group received EA at identical acupoints with skin penetration without stimulation.After 30 days,the ?nal body weight and coef?cients for the testis and epididymis were calculated and sperm parameters were measured.Immunohistochemical analyses were performed to detect expression of vimentin and α-tubulin in SCs and proliferating cell nuclear antigen(PCNA) immunoreactivity in germ cells.Apoptosis in germ cells was quanti?ed by the transferase biotin-dUTP nick end labeling assay.Results: Compared with the control group,the final body weight and testis/epididymis coefficients of rats in the TG-treated group were not significantly different,but the sperm count and motility were lower(P<0.05).Expressions of vimentin and α-tubulin were also signi?cantly weaker(P<0.01).The PCNA immunoreactivity of germ cells was decreased(P=0.059),whereas the apoptotic index of germ cells was increased signi?cantly(P<0.01).In contrast,EA at BL 23 and ST 36 acupoints signi?cantly improved the ?nal body weight as well as the sperm count,concentration and motility(P<0.01 or P<0.05).EA increased expression of vimentin and α-tubulin in SCs markedly,and signi?cantly enhanced PCNA immunoreactivity with decreased apoptosis in germ cells(P<0.01 or P<0.05).Conclusions: EA at BL 23 and ST 36 acupoints has protective effects on spermatogenesis in rats with OIKES.This effect seems to be achieved by attenuating TG-induced disruption of cytoskeletal protein in SCs.

Strong contact coupling of neuronal growth cones with height-controlled vertical silicon nanocolumns

In this study, we report that height-controlled vertically etched silicon nano- column arrays （vSNAs） induce strong growth cone-to-substrate coupling and accelerate in vitro neurite development while preserving the essential features of initial neurite formation. Large-scale preparation of vSNAs with flat head morphology enabled the generation of well-controlled topographical stimulation without cellular impalement. A systematic analysis on topography- induced variations on cellular morphology and cytoskeletal dynamics was conducted. In addition, neurite development on the grid-patterned vSNAs exhibited preferential adhesion to the nanostructured region and outgrowth directionality. The arrangement of cytoskeletal proteins and the expression of a focal adhesion complex indicated that a strong coupling existed between the underlying nanocolumns and growth cones. Furthermore, the height-controlled nanocolumn substrates differentially modulated neurite polarization and elongation. Our findings provide an important insight into neuron-nanotopography interactions and their role in cell adhesion and neurite development.