cell proliferation

摘要： Activating V600E in v-Raf murine sarcoma viral oncogene homolog B(BRAF)is a common driver mutation in cancers of multiple tissue origins,including melanoma and glioma.BRAF^(V600E) has also been implicated in neurodegeneration.The present study aims to characterize BRAF^(V600E) during cell death and proliferation of three major cell types of the central nervous system:neurons,astrocytes,and microglia.Multiple primary cultures(primary cortical mixed culture)and cell lines of glial cells(BV2)and neurons(SH-SY5Y)were employed.BRAF^(V600E) and BRAF^(WT) expression was mediated by lentivirus or retrovirus.Blockage of downstream effectors(extracellular signal-regulated kinase 1/2 and JNK1/2)were achieved by siRNA.In astrocytes and microglia,BRAF^(V600E) induces cell proliferation,and the proliferative effect in microglia is mediated by activated extracellular signal-regulated kinase,but not c-Jun N-terminal kinase.Conditioned medium from BRAF^(V600E)-expressing microglia induced neuronal death.In neuronal cells,BRAF^(V600E) directly induces neuronal death,through c-Jun N-terminal kinase but not extracellular signal-regulated kinase.We further show that BRAF-related genes are enriched in pathways in patients with Parkinson’s disease.Our study identifies distinct consequences mediated by distinct downstream effectors in dividing glial cells and in neurons following the same BRAF mutational activation and a causal link between BRAF-activated microglia and neuronal cell death that does not require physical proximity.It provides insight into a possibly important role of BRAF in neurodegeneration as a result of either dysregulated BRAF in neurons or its impact on glial cells.

摘要： Objective:Cleavage and polyadenylation specific factor 6(CPSF6)has been documented as an oncoprotein in different types of cancer.However,functions of CPSF6 have not been investigated yet in esophageal squamous cell carcinoma(ESCC).Here,we aimed to investigate the potential clinical values and biological functions of CPSF6 in ESCC.Methods:For determining the expression level of CPSF6 in ESCC patients,we analyzed published data,performed quantitative real-time polymerase chain reaction(RT-qPCR)and immunohistochemistry assays.Kaplan-Meier curves and log-rank tests were used for survival analyses.GO and KEGG analyses were done for CPSF6-related genes.Cell proliferation,colony formation and xenograft assays were conducted to verify the effects of CPSF6 on ESCC.In addition,cell cycle and apoptosis assays were also performed to manifest the functions of CPSF6 and circCPSF6.RNA pulldown and radioimmunoprecipitation(RIP)assays were used for confirming the interaction between circCPSF6(hsa_circ_0000417)and CPSF6 protein.The regulatory relationship between CPSF6 protein and circCPSF6 was determined by RT-qPCR.Results:We found that CPSF6 was upregulated in ESCC tissues and overexpression of cytoplasmic CPSF6 was associated with poor prognosis.GO and KEGG analyses suggested that CPSF6 could mainly affect cell division in ESCC.Further experiments manifested that CPSF6 promoted cell proliferation and colony formation in vitro.Xenograft assay showed that knockdown of CPSF6 significantly decreased tumor growth rate in vivo.Subsequently,we verified that depletion of CPSF6 led to cell cycle arrest and apoptosis.Finally,we validated that CPSF6,as a circRNA-binding protein,interacted with and regulated its circular isoform circCPSF6(hsa_circ_0000417),of which depletion also resulted in cell cycle arrest and cell apoptosis in ESCC.Conclusions:These findings gave us insight that overexpression of cytoplasmic CPSF6 protein is associated with poor prognosis in ESCC and CPSF6 may function as an oncoprotein,at least in part,through regulating circCPSF6 expression.

摘要： Hepatocytes can divide rapidly and proliferate in the absence of inflammation and fibroplasia in the damaged or partial hepatectomy(PHx)of the liver,which is essential for the recovery of related patients.Recent studies have found that bile acids(BA)play an important role in the process of liver regeneration.In the early stages of PHx,bile acid overload occurs,and liver injury is aggravated by loading.Later bile acids can induce protective and proliferative responses in the liver and promote liver regeneration.In this paper,we summarize the negative effects after bile acid overload and its positive role as a signaling molecule involved in related signaling pathways on liver regeneration,including protection of the liver and promotion of liver regeneration,and its double-edswordged"in Liver regeneration.This provides a theoretical basis for subsequent in-depth study of the mechanism and benefit avoidance in clinical treatment.

摘要： Objective:To explore the anti-glioma effect and influence of luteolin through mi R-384/PIWIL4 pathway.Methods:30 cases of human glioblastoma multiform U251,U87,A172,SHG44 cells,and 30 cases of the normal glial cell NHA were treated with luteolin.The control group was detected by Western blot,and the level of PIWIL4 in the cytoplasm and nucleus of each cell line was analyzed.The expression levels of PIWIL4 m RNA and mi R-384 in each cell line were detected by qRT-PCR.Results:the glial cell NHA with normal PIWIL4 level in the cytoplasm was related to U251,U87,A172,and SHG44 human pleomorphic glioma cells.The m RNA expression levels of U251,U87,A172,and SHG44 human pleomorphic glioma cells were significantly higher than those of normal gliomas(P<0.05),and the mi R-384 expression levels of U251,U87,A172,and SHG44 human pleomorphic glioma cells were significantly higher than those of normal gliomas(P<0.05).With the continuous increase of luteolin concentration,glioma cells showed a significant increase in dependence(P<0.05).Conclusion:Luteolin could up-regulate mi R-384,inhibit PIWIL4,and regulate the biological activity of glioma cell proliferation and migration.

摘要： Ulcerative colitis(UC)is an incurable and highly complex digestive disease affecting millions of people worldwide.Compared to the current therapeutic drugs,bioactive peptides are more promising and safe substances as functional foods or drugs for the prevention and treatment of UC.The alcohol-soluble components from fermentation broth by fresh wheat germ and apple(AC-WGAF)were found to be effective in UC prevention in dextran sulfate sodium-induced mice in vivo.Herein,4 novel peptides are identifi ed from AC-WGAF by membrane ultrafi ltration,recycling preparative high-performance liquid chromatography,and matrix-assisted laser desorption–ionization time-of-fl ight/time-of-fl ight mass spectrometry,possessing anticolitis activity via using an in vitro model.One of those peptides named T24(PVLGPVRGPFPLL)exhibited the most remarkable anti-colitis activity by preventing tight junction protein loss,maintaining epithelial barrier integrity,and promoting cell proliferation during in vitro and in vivo studies by regulating mitogen-activated protein kinase signaling pathways.Thus,T24 is a promising peptide as a functional food or novel drug for UC prevention and treatment.

摘要： The COP9 signalosome(CSN)is a highly conserved protein complex composed of 8 subunits(CSN1 to CSN8).The individual subunits of the CSN play essential roles in cell proliferation,tumorigenesis,cell cycle regulation,DNA damage repair,angiogenesis,and microenvironmental homeostasis.The CSN complex has an intrinsic metalloprotease that removes the ubiquitin-like activator NEDD8 from cullin-RING ligases(CRLs).Binding of neddylated CRLs to CSN is sensed by CSN4 and communicated to CSN5 with the assistance of CSN6,thus leading to the activation of deneddylase.Therefore,CSN is a crucial regulator at the intersection between neddylation and ubiquitination in cancer progression.Here,we summarize current understanding of the roles of individual CSN subunits in cancer progression.Furthermore,we explain how the CSN affects tumorigenesis through regulating transcription factors and the cell cycle.Finally,we discuss individual CSN subunits as potential therapeutic targets to provide new directions and strategies for cancer therapy.

摘要： BACKGROUND Esophageal squamous cell carcinoma(ESCC)is one of the most common malignancies.A total of 45 kinesin superfamily proteins(KIFs)have been identified in humans,among which several family members have demonstrated varied functions in tumor pathobiology via different mechanisms,including regulation of cell cycle progression and metastasis.KIFC3 has microtubule motor activity and is involved in cancer cell invasion and migration,as well as survival.However,the role of KIFC3 in ESCC is still unknown.AIM To evaluate the role of KIFC3 in ESCC and the underlying mechanisms.METHODS Expression of KIFC3 was evaluated in ESCC tissues and adjacent normal esophageal tissues.The prognostic value of KIFC3 was analyzed using Kaplan-Meier Plotter.Colony formation,EdU assays,cell cycle analysis,Transwell assay,immunofluorescence,and western blotting were performed in ESCC cell lines after transfection with pLVX-Puro-KIFC3-shRNA-and pLVX-Puro-KIFC3-expressing lentiviruses.A xenograft tumor model in nude mice was used to evaluate the role of KIFC3 in tumorigenesis.Inhibitor ofβ-catenin,XAV-939,was used to clarify the mechanism of KIFC3 in ESCC.To analyze the differences between groups,t test and nonparametric tests were used.P<0.05 was considered statistically significant.RESULTS Immunohistochemical staining indicated that KIFC3 was upregulated in ESCC tissues compared with adjacent normal tissues.Kaplan-Meier Plotter revealed that overexpressed KIFC3 was associated with poor prognosis in ESCC patients.Colony formation and EdU assay showed that KIFC3 overexpression promoted cell proliferation,while KIFC3 knockdown inhibited cell proliferation in ESCC cell lines.In addition,cell cycle analysis showed that KIFC3 overexpression promoted cell cycle progression.KIFC3 knockdown suppressed ESCC tumorigenesis in vivo.Transwell assay and western blotting revealed that KIFC3 overexpression promoted cell migration and invasion,as well as epithelial-mesenchymal transition(EMT),while KIFC3 knockdown showed the opposite results.Mechanistically,KIFC3 overexpression promoted β-catenin signaling in KYSE450 cells;however,the role of KIFC3 was abolished by XAV-939,the inhibitor of β-catenin signaling.CONCLUSION KIFC3 was overexpressed in ESCC and was associated with poor prognosis.Furthermore,KIFC3 promoted proliferation,migration and invasion of ESCC via β-catenin signaling and EMT.

摘要： Objective Long non-coding RNAs(lncRNAs)regulate tumor development and progression by promoting tumor proliferation,invasion,and metastasis.The aim of the study was to investigate the effects of lncRNA growth arrest-special 5(GAS5)on proliferation and apoptosis of hepatocellular carcinoma(HCC)cells through miR-26a-5p action.Methods Expression levels of GAS5 were detected in cancerous and paracancerous tissue of 80 HCC patients by RT-qPCR.The starBase tool predicted that GAS5 had binding sites for the miRNA miR-26a-5p,which was also highly expressed in HCC tissue.The relationship between GAS5 and miR-26a-5p was confirmed using a luciferase reporter assay.The role of these lncRNAs was further explored by transfecting plasmids into SMMC-7721 cells and classifying the cells as follows:NC group,GAS5 group,anti-miR-26a-5p group,and GAS5+miR-26a-5p group.Cell proliferation,cell cycle,and apoptosis were detected in each group.The relationship between miR-26a-5p and phosphatase and tensin homolog deleted on chromosome 10(PTEN)was analyzed by TargetScan database prediction and luciferase reporter assay.Western blotting was used to quantify PTEN,phosphatidylinositol 3-kinase(PI3K),phosphorylated protein kinase B(p-Akt),cyclin D1,and human P27 protein(P27).Results GAS5 was downregulated,while miR-26a-5p was upregulated in HCC tissue compared to in paracancerous tissue.High GAS5 levels and low miR-26a-5p levels inhibited cell proliferation,increased the number of G0/G1 phase cells,promoted cell apoptosis,promoted PTEN and P27 expression,and inhibited PI3K,P-Akt,and cyclin D1 expression at the protein level.Upregulation of miR-26a-5p attenuated the effects of GAS5 upregulation on the proliferation,cell cycle,and apoptosis of HCC cells and on the expression of PTNE/PI3K/Akt signaling pathway-related proteins.Conclusion Low GAS5 levels regulate the proliferation and apoptosis of HCC cells via the PTNE/PI3K/Akt signaling pathway and are linked to upregulation of miR-26a-5p.

摘要： In order to explore the role of forkhead box protein O1(FoxO1)in the lipid metabolism and cell proliferation,goose primary hepatocytes were isolated and incubated with insulin or PI3K-Akt-mTOR pathway dual inhibitor NVPBEZ235,and then transfected with FoxO1 interference plasmid.The related parameters of lipid metabolism and cell proliferation were measured.The results firstly showed that FoxO1 interference increased the intracellular TG and lipids concentration(P0.05).Lastly,the decrease of intracellular TG and lipids concentration and PI induced by NVP-BEZ235 was up-regulated by FoxO1 interference significantly(P<0.05).In summary,FoxO1 could regulate the lipids metabolism and cell proliferation mediated by PI3K-Akt-mTOR signaling pathway in goose primary hepatocytes.Further investigations are required to highlight the potential role of FoxO1 in the lipid metabolism and cell proliferation mediated by insulin in goose primary hepatocyte.

摘要： Korean pine is an important afforestation tree species in Northeast China,which has a high ecological and economic value.Although regeneration of somatic embryogenesis using immature zygotic embryos of Korean pine as explants has been successful,it cannot be applied to automation and large-scale production.Therefore,we urgently need a method that can increase the output of somatic embryos(SEs)to meet the needs of large-scale production.We used Korean pine 1-1 and 1-100 cell lines as research materials to evaluate the effects of inoculum-density,culture time,orbiting speed,vessel volume,plant growth regulator(PGR)concentration,and carbon source on the proliferation of embryogenic tissue(ET).The somatic embryogenesis ability of ET cultured in different liquid suspension media was also evaluated.We found that during liquid suspension culture of Korean pine ET,the sedimented cell volume(SCV),fresh weight(FW)and dry weight(DW)were affected by inoculumdensity,culture time,orbiting speed,2,4-D concentration,6-BA concentration and carbon source type.Fourty mg⋅mL^(−1)ET were transferred to a 200 mL Erlenmeyer flask containing 20 mL liquid medium,and cultured at 100 rpm/min for 14 days to obtain the maximum proliferation.In addition,we also found that SCV,FW and DW were higher when PGRs were reduced in the liquid suspension medium.The substitution of maltose for sucrose resulted in slow growth of cultures and limited SE yield(13 SEs g^(−1)FW).Although culture proliferation was high at 50 rpm,SE yield was inhibited by 48%compared with 100 rpm(50 rpm=33 SEs g^(−1)FW;100 rpm/min=70 SEs g^(−1)FW).Cultivation in low-concentration PGR(1.15μM⋅L^(−1)2,4-D,0.25μM⋅L^(−1)6-BA)and sucrose liquid medium at 100 rpm/min(80 SEs g^(−1)FW)could not only promote culture proliferation but also increase SE yield.The determination of the suspension culture scheme of Korean pine ET provides a reference for further expansion to bioreactor culture in the future and lays a foundation for the automation and scale of somatic embryogenesis of Korean pine.