Neurofibromatosis type 1(NF1)is a genetic disorder affecting 1 in 3000 people due to heterozygous mutations in the NF1 gene.Patients with NF1 can develop multiple symptoms,such as neurofibromas,skin hyperpigmentation,...Neurofibromatosis type 1(NF1)is a genetic disorder affecting 1 in 3000 people due to heterozygous mutations in the NF1 gene.Patients with NF1 can develop multiple symptoms,such as neurofibromas,skin hyperpigmentation,and bone abnormalities,including tibial pseudarthrosis and spine deformity.Here,we aimed to elucidate the cellular origin and pathogenic mechanism of NF1 spine deformity.We explored the Prss56-Nf1 knockout(KO)mouse model that recapitulates neurofibromas and pseudarthrosis by carrying Nf1 gene inactivation in Prss56-expressing boundary cap cells,a neural crest subset,and their derivatives.Micro-CT analyses showed that Prss56-Nf1 KO mice exhibit spine deformity from 12 months of age,associated with vertebral anomalies reminiscent of patients with NF1.Fate mapping revealed a significant increase in OSX^(+)osteoblasts of the Prss56 lineage in vertebrae of Prss56-Nf1 KO mice.Increased traced Nf1-deficient cells correlated with increased vertebral bone volume and kyphosis spine curvature.Finally,we showed that treating Prss56-Nf1 KO mice with RAS-MAPK pathway inhibitors prevented spine deformity.Overall,the Prss56-Nf1 KO mouse model unravels the role of osteoblasts from the Prss56 lineage as the cellular origin of NF1 spine deformity and highlights RAS-MAPK pathway inhibition as a promising therapeutic strategy for preventing NF1 spine deformity.展开更多
Oncogenic KRAS mutations drive metabolic reprogramming in pancreatic ductal adenocarcinoma(PDAC).Src-homology 2 domaincontaining phosphatase 2(SHP2)is essential for full KRAS activity,and promising dual SHP2/mitogen-a...Oncogenic KRAS mutations drive metabolic reprogramming in pancreatic ductal adenocarcinoma(PDAC).Src-homology 2 domaincontaining phosphatase 2(SHP2)is essential for full KRAS activity,and promising dual SHP2/mitogen-activated protein kinase(MAPK)inhibition is currently being tested in clinical trials.Exploitable metabolic adaptations may contribute to invariably evolving resistance.To understand the metabolic changes induced by dual inhibition,we comprehensively tested human and murine PDAC cell lines,endogenous tumor models,and patient-derived organoids,which are representative of the full spectrum of PDAC molecular subtypes.We found that dual SHP2/mitogen-activated protein kinase kinase(MEK1/2)inhibition induces major alterations in mitochondrial mass and function,impacts reactive oxygen species(ROS)homeostasis and triggers lipid peroxidase dependency.Anabolic pathways,autophagy and glycolysis were also profoundly altered.However,most strikingly,mitochondrial remodeling was evident,persisting into a therapy-resistant state.The resulting vulnerability to the induction of ferroptotic cell death via the combination of vertical SHP2/MEK1/2 with glutathione peroxidase(GPX4)inhibition was largely independent of the PDAC molecular subtype and was confirmed with direct targeting of RAS.The triple combination of SHP2/MEK1/2 inhibition and the ferroptosis-inducing natural compound withaferin A suppressed tumor progression in an endogenous PDAC tumor model in vivo.Our study offers a metabolic leverage point to reinforce RAS pathway interference for targeted PDAC treatment.展开更多
Objective:To characterize the tumor-suppressive role of LINC00936 in non-small cell lung cancer(NSCLC)through mechanistic exploration of its regulatory pathways.Methods:Bioinformatics interrogation of TCGA/NSCLC cohor...Objective:To characterize the tumor-suppressive role of LINC00936 in non-small cell lung cancer(NSCLC)through mechanistic exploration of its regulatory pathways.Methods:Bioinformatics interrogation of TCGA/NSCLC cohorts assessed LINC00936 expression,clinical correlations,and immune contexture.Functional enrichment analyses predicted pathway associations.In H1299 cells,LINC00936 overexpression(plasmid)and knockdown(siRNA)models were validated by RT-qPCR.Transcriptomic profiling identified differentially expressed genes(DEGs)subjected to KEGG pathway analysis.Results:LINC00936 was significantly downregulated in NSCLC tissues(TCGA,P<0.05)and cell lines(vs.16-HBE,P<0.05),correlating with poor prognosis and altered tumor-infiltrating immune subsets.DEG enrichment implicated Ras/MAPK signaling as the dominant pathway(FDR<0.05).Successful LINC00936 modulation(overexpression/knockdown,P<0.05)confirmed its regulatory capacity.Conclusion:LINC00936 acts as a tumor suppressor in NSCLC via Ras/MAPK pathway modulation,proposing its therapeutic candidacy for precision oncology strategies.展开更多
Background:Glioma is the most common tumor of the central nervous system with a poor prognosis.This study aims to explore the role of calcium/calmodulin-dependent protein kinase IIβ(CAMK2B)in regulating the malignant...Background:Glioma is the most common tumor of the central nervous system with a poor prognosis.This study aims to explore the role of calcium/calmodulin-dependent protein kinase IIβ(CAMK2B)in regulating the malignant progression of glioma cells,as well as the molecular mechanisms underlying these malignant behaviors.Methods:The correlation between CAMK2B expression in gliomas and patient prognosis was analyzed using immunohistochemistry,quantitative reverse transcription polymerase chain reaction(qRT-PCR),and western blot.Furthermore,the study explored the role of CAMK2B in glioma cell proliferation,invasion,and migration using cell counting kit-8(CCK-8),5-Ethynyl-2′-deoxyuridine(EdU),wound healing,transwell,and in vivo tumor xenograft assays.Result:Patients with high CAMK2B expression exhibited significantly better prognostic outcomes compared to those with low expression levels.Furthermore,CAMK2B expression was significantly lower in glioma tissues and cells compared to both normal brain tissue and human astrocyte cell lines.Notably,overexpression of CAMK2B in glioma cells led to an approximate 40%reduction in proliferative capacity and a 60–70%decrease in invasive and migratory abilities,compared to control glioma cells.These differences were statistically significant at p<0.05.Conversely,knockdown of CAMK2B using siRNA-CAMK2B significantly enhanced the proliferative,invasive,and migratory capabilities of glioma cells in both in vitro and in vivo settings,enhancing these abilities by 1.5 to 3 times.Notably,these effects were reversed through the application of the Rat Sarcoma viral oncogene homolog(Ras)pathway inhibitor,Salirasib.Western blot analysis revealed that knockdown of CAMK2B led to activation of the Ras/Rapidly Accelerated Fibrosarcoma(Raf)/Mitogen-activated protein kinase kinase(MEK)/Extracellular signal-regulated kinase(ERK)signaling pathway in glioma cell lines,whereas overexpression of CAMK2B resulted in the suppression of this pathway.Conclusion:CAMK2B inhibits glioma proliferation,invasion,andmigration through the Ras/Raf/MEK/ERK signaling pathway.展开更多
Nasopharyngeal carcinoma(NPC)is the third most common malignancy with a high recurrence and metastasis rate in South China.Natural compounds extracted from traditional Chinese herbal medicines have been developed and ...Nasopharyngeal carcinoma(NPC)is the third most common malignancy with a high recurrence and metastasis rate in South China.Natural compounds extracted from traditional Chinese herbal medicines have been developed and utilized for the treatment of a variety of cancers with modest properties and slight side effects.Maackiain(MA)is a type of flavonoid that was first isolated from leguminous plants,and it has been reported to relieve various nervous system disorders and exert anti-allergic as well as antiinflammatory effects.In this study,we demonstrated that MA inhibited proliferation,arrested cell cycle and induced apoptosis in nasopharyngeal carcinoma CNE1 and CNE2 cells in vitro and in vivo.The expression of the related proteins associated with these processes were consistent with the above effects.Moreover,transcriptome sequencing and subsequent Western blot experiments revealed that inhibition of the MAPK/Ras pathway may be responsible to the anti-tumor effect of MA on NPC cells.Therefore,the effects of MA and an activator of this pathway,tertiary butylhydroquinone(TBHQ),alone or combination,were investigated.The results showed TBHQ neutralized the inhibitory effects of MA.These data suggest that MA exerts its anti-tumor effect by inhibiting the MAPK/Ras signaling pathway and it has the potential to become a treatment for patients with NPC.展开更多
Objective To assess the effects of Yi Qi Jie Du Formula(YQJDF)combined with salinomycin(SAL)on nasopharyngeal carcinoma stem cells(NPC-SCs)and investigate the underlying molecular mechanisms.Methods Cell counting meth...Objective To assess the effects of Yi Qi Jie Du Formula(YQJDF)combined with salinomycin(SAL)on nasopharyngeal carcinoma stem cells(NPC-SCs)and investigate the underlying molecular mechanisms.Methods Cell counting methods,the CCK-8 assay,transwell migration assay and JC-1 staining,were used to observe the effects of the combination on the proliferation,migration and apoptosis of NPC stem cells,respectively.Western blot was used to detect the levels of protein in NPC-SCs.Results YQJDF combined with SAL had a synergistic effect on the inhibition of proliferation and migration and induction of NPCSC apoptosis.Mechanistically,YQJDF combined with SAL synergistically upregulated the levels of apoptotic proteins,including cleaved Caspase-3,cleaved Caspase-7 and cleaved Caspase-9.Moreover,YQJDF combined with SAL synergistically decreased the levels of CD44,p-c-Src,Ras,p-PKCδ,p-MEK,p-c-Raf,p-ERK1/2 and p-AKT proteins.Conclusions The combination of YQJDF and SAL has a synergistic effect on the inhibition of NPC-SC proliferation and migration and induction of apoptosis,which may be closely related to the downregulation of the CD44/Ras signaling pathway.展开更多
An increasing number of studies report that the Ras/Raf/extracellular signal-regulated kinase 1/2 （ERK1/2） signaling pathway has a death-promoting apoptotic function in neural cells. We hypothesized that the Ras/Raf...An increasing number of studies report that the Ras/Raf/extracellular signal-regulated kinase 1/2 （ERK1/2） signaling pathway has a death-promoting apoptotic function in neural cells. We hypothesized that the Ras/Raf/ERK1/2 signaling pathway may be abnormally regulated in rat injured spinal cord models. The weight drop method was used to establish rat spinal cord injury at T9. Western blot analysis and immunohistochemical staining revealed Ras expression was dramatically elevated, and the phosphorylations of A-Raf, B-Raf and C-Raf were all upregulated in the injured spinal cord. Both mitogen-activated protein kinase kinase 1/2 and ERK1/2, which belong to the Ras/Raf signaling kinases, were upregulated. These results indicate that Ras/Raf/ ERK1/2 signaling may be upregulated in injured spinal cord and are involved in recovery after spinal cord injury.展开更多
Objective:To investigate the effect of Bushen Antai Granule on the mRNA and protein expression of Ras protein/mitogen activated protein kinase mediated by vascular endothelial growth factor receptor 2 with small inter...Objective:To investigate the effect of Bushen Antai Granule on the mRNA and protein expression of Ras protein/mitogen activated protein kinase mediated by vascular endothelial growth factor receptor 2 with small interference RNA interference.Methods:Method to construct the placenta microvascular endothelial cells,and the preparation of kidney fetus granule drug-containing serum,select the best drug-containing serum concentration,it can be divided into normal group,the serum siRNA-NC normal serum group,drug serum,siRNA normal serum group,siRNA drug serum group,using real-time fluorescent quantitative PCR,Western blotting,immunofluorescence test respectively the RAS/MAPK mRNA and protein expression.Results:Results there was no significant difference in Ras and MAPK mRNA and protein expression between the normal group and the negative control group(P>0.05).The mRNA and protein expressions of Ras and MAPK in the drug serum group were significantly higher than those in the normal serum group(P<0.01).Ras and MAPK mRNA and protein expression were significantly decreased in siRNA1 normal serum group compared with normal serum group(P<0.01).Ras,MAPK mRNA and protein expression in siRNA1 drug serum group were significantly different from that in siRNA1 normal serum group(P<0.01).Conclusion:Conclusion The therapeutic effect of Bushen Antai Granule on recurrent abortion may be realized by upregulation of RAS/MAPK mRNA and protein expression.展开更多
[Objectives]To explore the neuroprotective effects and mechanism of Longan Aril(LA)effective parts on PC12 cells injured by H2O2.[Methods]The neuroprotective effects of LA were evaluated by the cell viability,SOD and ...[Objectives]To explore the neuroprotective effects and mechanism of Longan Aril(LA)effective parts on PC12 cells injured by H2O2.[Methods]The neuroprotective effects of LA were evaluated by the cell viability,SOD and MDA content,apoptosis assay and relative protein expression of Aβand p-Tau.The neuroprotective mechanism of LA was studied by using metabolomics and network pharmacology,and the expressions of RAS/MEK/ERK signaling pathway-related proteins were detected by western blotting.[Results]LA could improve the cell survival rate and SOD content,and reduce apoptosis and expression of Aβand p-tau.Inhibition of RAS/MEK/ERK signaling pathway is a possible mechanism of LA neuroprotective effects.[Conclusions]LA has a neuroprotective effects in vitro and be likely to inhibit the process of AD by inhibition of RAS/MEK/ERK signalling pathway.展开更多
Spaceflight-associated immune system weakening ultimately limits the ability of humans to expand their presence beyond the earth's orbit. A mechanistic study of microgravity-regulated immune cell function is neces...Spaceflight-associated immune system weakening ultimately limits the ability of humans to expand their presence beyond the earth's orbit. A mechanistic study of microgravity-regulated immune cell function is necessary to overcome this challenge. Here, we demonstrate that both spaceflight (real) and simulated microgravity significantly reduce macrophage differentiation, decrease macrophage quantity and functional polarization, and lead to metabolic reprogramming, as demonstrated by changes in gene expression profiles. Moreover, we identified RAS/ERK/NFκB as a major microgravity-regulated pathway. Exogenous ERK and NFκB activators significantly counteracted the effect of microgravity on macrophage differentiation. In addition, microgravity also affects the p53 pathway, which we verified by RT-qPCR and Western blot. Collectively, our data reveal a new mechanism for the effects of microgravity on macrophage development and provide potential molecular targets for the prevention or treatment of macrophage differentiation deficiency in spaceflight.展开更多
Stromal cell-derived factor-1 and its receptor C-X-C chemokine receptor 4(CXCR4) have been shown to regulate neural regeneration after stroke.Howeve r,whether stromal cell-derived factor-1 receptor CXCR7,which is wide...Stromal cell-derived factor-1 and its receptor C-X-C chemokine receptor 4(CXCR4) have been shown to regulate neural regeneration after stroke.Howeve r,whether stromal cell-derived factor-1 receptor CXCR7,which is widely distributed in the develo ping and adult central nervous system,participates in neural regeneration remains poorly unde rstood.In this study,we established rat models of focal cerebral ischemia by injecting endothelin-1 into the cerebral co rtex and striatum.Starting on day 7 after injury,CXCR7-neutralizing antibody was injected into the lateral ventricle using a micro drug delivery system for 6 consecutive days.Our results showed that CXCR7-neutralizing antibody increased the total length and number of sprouting co rticospinal tra ct fibers in rats with cerebral ischemia,increased the expression of vesicular glutamate transporter 1 and growth-related protein 43,marke rs of the denervated spinal cord synapses,and promoted the differentiation and maturation of oligodendrocyte progenitor cells in the striatum.In addition,CXCR7 antibody increased the expression of CXCR4 in the striatum,increased the protein expression of RAS and ERK1/2 associated with the RAS/ERK signaling pathway,and im proved rat motor function.These findings suggest that CXCR7 improved neural functional recovery after ischemic stroke by promoting axonal regeneration,synaptogenesis,and myelin regeneration,which may be achieved by activation of CXCR4 and the RAS/ERK1/2 signaling pathway.展开更多
Abnormal activation of the Ras/Raf/Mek/Erk signaling cascade plays an important role in glioma. Inhibition of this aberrant activity could effectively hinder glioma cell proliferation and promote cell apoptosis. To in...Abnormal activation of the Ras/Raf/Mek/Erk signaling cascade plays an important role in glioma. Inhibition of this aberrant activity could effectively hinder glioma cell proliferation and promote cell apoptosis. To investigate the mechanism of gJioblastoma treatment by neural stem ceiJ trans- plantation with respect to the Ras/Raf/Mek/Erk pathway, C6 glioma cells were prepared in sus- pension and then infused into the rat brain to establish a glioblastoma model. Neural stem cells isolated from fetal rats were then injected into the brain of this glioblastoma model. Results showed that Raf-1, Erk and Bcl-2 protein expression significantly increased, while Caspase-3 protein expression decreased. After transplantation of neural stem cells, Raf-1, Erk and Bcl-2 protein expression significantly decreased, while Caspase-3 protein expression significantly in-creased. Our findings indicate that transplantation of neural stem cells may promote apoptosis of glioma cells by inhibiting Ras/Raf/Mek/Erk signaling, and thus may represent a novel treatment approach for glioblastoma.展开更多
BACKGROUND： Electromagnetic radiation can influence dopamine （DA） synthesis in brain tissues or ceils, but electromagnetic frequencies, intensities, and radiation time can produce different effects. In addition, th...BACKGROUND： Electromagnetic radiation can influence dopamine （DA） synthesis in brain tissues or ceils, but electromagnetic frequencies, intensities, and radiation time can produce different effects. In addition, the signal pathway by which electromagnetic radiation influences DA synthesis remains controversial. OBJECTIVE： To determine tyrosine hydroxylase （TH） expression in PC12 cells and DA levels in cell culture media after different periods of low-frequency pulsed electric field （LF-PEF） stimulation, and to determine how LF-PEF signaling stimulates TH synthesis using inhibitors. DESIGN, TIME AND SETTING： A parallel, controlled, cell experiment was performed at the Laboratory of Cell Biology, School of Life Science, East China Normal University, between January and October 2006. MATERIALS： PC12 cells were purchased from the Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, China. Nerve growth factor was purchased from PeproTech, USA. The protein kinase A inhibitor, H-89, and mitogen-activated protein kinase kinase inhibitor, U0126, were purchased from Sigma, USA. METHODS： （1） Following routine culture in Dulbecco＇s modified eagle medium, primary PC12 cells were stimulated under LF-PEF （pulse frequency 50.Hz, pulse width 20 μs, peak field strength 1 V/m） for 5, 10, 15, 20, and 30 minutes. （2） Inhibitors （H-89 or U0126, 1 μmol/L） were added 30 minutes before LF-PEF stimulation for 10 minutes. MAIN OUTCOME MEASURES： （1） TH expression was determined by Western blot in PC12 cells at 0.5, 1,2, 3, and 4 days after LF-PEF stimulation. Similarly, DA was measured by high-performance liquid chromatography in media at 2, 3, 4, or 5 days after LF-PEE （2） TH expression was detected 1 day after H-89 or U0126 treatment and LF-PEE RESULTS： （1） Short-term LF-PEF stimulation （5 and 10 minutes） increased TH expression and media DA levels after short-term culture （2 days） （P 〈 0.01）, but both parameters decreased with longer culture （3 4 days） （P 〈 0.01）. Long-term LF-PEF stimulation （15, 20, or 30 minutes） decreased TH and DA synthesis, followed by a rapid increase （P 〈 0.01）. （2） H89 could completely inhibit TH expression in PC12 cells stimulated by LF-PEF for 10 minutes, while the inhibition rate of U0126 was 53.2%. CONCLUSION： Short-term LF-PEF first promotes then inhibits, while long-term LF-PEF first inhibits then promotes, TH and DA synthesis. LF-PEF stimulation regulates TH expression primarily by activating protein kinase A to regulate DA synthesis.展开更多
Subject Code:H16With the support by the National Natural Science Foundation of China,a collaborative study by the research groups led by Prof.Xiao Zhixiong(肖智雄)from the College of Life Science,Sichuan University de...Subject Code:H16With the support by the National Natural Science Foundation of China,a collaborative study by the research groups led by Prof.Xiao Zhixiong(肖智雄)from the College of Life Science,Sichuan University demonstrates thatΔNp63αis a common inhibitory target in oncogenic PI3K/Ras/Her2-induced展开更多
Tetrabromobisphenol A(TBBPA)is a flame retardant that adversely affects the environment and human health.The present study exposed HepG2 cells to low concentrations of TBBPA daily to investigate the changes in gene re...Tetrabromobisphenol A(TBBPA)is a flame retardant that adversely affects the environment and human health.The present study exposed HepG2 cells to low concentrations of TBBPA daily to investigate the changes in gene regulation,mainly related to pathways associated with the endocrine system.The quantitative polymerase chain reaction(qPCR)confirmed that prolonged exposure gradually activated the thyroid hormone and parathyroid hormone signaling pathways.The expression levels of genes related to the thyroid hormone signaling pathway were upregulated(1.15-8.54 times)after five generations of exposure to 1 and 81 nM TBBPA.Furthermore,co-exposure to 81 nM TBBPA and 0.5 nM thyroid hormone receptor antagonist for five generations significantly reduced the expression of thyroid hormone and parathyroid hormone receptors.Meanwhile,81 nM TBBPA inhibited the activation of the Ras pathway and downregulated Ras gene expression level(3.7 times),indicating the association between the toxic effect and thyroid hormone receptors.Additionally,our experiments revealed that the thyroid hormone pathway regulated the induction of the Ras signaling pathway by TBBPA.The study thus proves that daily exposure to TBBPA interferes with the thyroid hormone signaling pathway and subsequently the endocrine system.展开更多
Growth of the Pacific white shrimp Litopenaeus vannamei,the most important farmed crustacean,has consistently been a focal point for breeders.Over the past decades,some candidate genes for shrimp growth have been iden...Growth of the Pacific white shrimp Litopenaeus vannamei,the most important farmed crustacean,has consistently been a focal point for breeders.Over the past decades,some candidate genes for shrimp growth have been identified.However,fur-ther research is needed to elucidate the molecular regulatory mechanism of these genes.LvMmd2 was previously identified as a candidate gene that may inhibit the growth of L.vannamei.In this study,we analyzed the genotype and expression of the LvMmd2 gene in a breeding family and indicated its role as a growth-inhibiting gene.We found that LvMmd2 co-localized with its homolog LvPAQR3 at the Golgi apparatus.Using co-immunoprecipitation(Co-IP)and DUAL membrane system yeast two-hybrid(MbY2H),we indicated the interactions between LvMmd2 and LvPAQR3,LvPAQR3 and LvRaf1,as well as LvMmd2 and LvRho.These results suggest that LvMmd2 directly and indirectly regulates the Ras signaling pathway.Furthermore,we show that the LvMmd2 gene may indirectly affect the PI3K/AKT,insulin,and Hippo signaling pathways to regulate cell proliferation and differentiation via LvPAQR3 and LvRaf1.Through transcriptome and MbY2H analyses,we have also revealed the interaction between LvMmd2 and proteins involved in growth,immunity,protein transport,synthesis,and modification.These findings demonstrate the various molecular pathways through which LvMmd2 regulates L.vannamei growth.This study provides insights into the mechanism of shrimp growth regulated by Mmd2,enhances our understanding of LvMmd2 function,and highlights its potential application in shrimp breeding.展开更多
RNA modifications affect many biological processes and physiological diseases.The 5-methylcytosine(m^(5)C)modification regulates the progression of multiple tumors.However,its characteristics and functions in hepatoce...RNA modifications affect many biological processes and physiological diseases.The 5-methylcytosine(m^(5)C)modification regulates the progression of multiple tumors.However,its characteristics and functions in hepatocellular carcinoma(HCC)remain largely unknown.Here,we found that HCC tissues had a higher m^(5)C methylation level than the adjacent normal tissues.Transcriptome analysis revealed that the hypermethylated genes mainly participated in the phosphokinase signaling pathways,such as the Ras and PI3K-Akt pathways.The m^(5)C methyltransferase NSUN2 was highly expressed in HCC tissues.Interestingly,the expression of many genes was positively correlated with the expression of NSUN2,including GRB2,RNF115,AATF,ADAM15,RTN3,and HDGF.Real-time PCR assays further revealed that the expression of the mRNAs of GRB2,RNF115,and AATF decreased significantly with the down-regulation of NSUN2 expression in HCC cells.Furthermore,NSUN2 could regulate the cellular sensitivity of HCC cells to sorafenib via modulating the Ras signaling pathway.Moreover,knocking down NSUN2 caused cell cycle arrest.Taken together,our study demonstrates the vital role of NSUN2 in the progression of HCC.展开更多
基金Association Neurofibromatoses et Recklinghausen(C.C)Agence Nationale de la Recherche-18-CE14-0033(C.C.,P.T.)+2 种基金Agence Nationale de la Recherche-21-CE18-007-01(C.C.,P.T.)US Department of the Army NF220019(C.C.)supported by a PhD fellowship from the University Paris-Est Créteil and the Fondation pour la Recherche Médicale(FRM)(FDT202304016600 to FK and ECO202306017399 to CG)。
文摘Neurofibromatosis type 1(NF1)is a genetic disorder affecting 1 in 3000 people due to heterozygous mutations in the NF1 gene.Patients with NF1 can develop multiple symptoms,such as neurofibromas,skin hyperpigmentation,and bone abnormalities,including tibial pseudarthrosis and spine deformity.Here,we aimed to elucidate the cellular origin and pathogenic mechanism of NF1 spine deformity.We explored the Prss56-Nf1 knockout(KO)mouse model that recapitulates neurofibromas and pseudarthrosis by carrying Nf1 gene inactivation in Prss56-expressing boundary cap cells,a neural crest subset,and their derivatives.Micro-CT analyses showed that Prss56-Nf1 KO mice exhibit spine deformity from 12 months of age,associated with vertebral anomalies reminiscent of patients with NF1.Fate mapping revealed a significant increase in OSX^(+)osteoblasts of the Prss56 lineage in vertebrae of Prss56-Nf1 KO mice.Increased traced Nf1-deficient cells correlated with increased vertebral bone volume and kyphosis spine curvature.Finally,we showed that treating Prss56-Nf1 KO mice with RAS-MAPK pathway inhibitors prevented spine deformity.Overall,the Prss56-Nf1 KO mouse model unravels the role of osteoblasts from the Prss56 lineage as the cellular origin of NF1 spine deformity and highlights RAS-MAPK pathway inhibition as a promising therapeutic strategy for preventing NF1 spine deformity.
基金This study was funded by the German Cancer Aid,Deutsche Krebshilfe,Project ID 70113697(to D.A.R.)the German Research Foundation,Deutsche Forschungsgemeinschaft(DFG),CRC1479(Project ID 441891347,P17 to D.A.R.,P01 to R.Z.,P06 to O.G.,S01 to M.B.and O.S.,S02 to W.R.)+10 种基金supported by the DFG,within CRC1160(Project ID 256073931,Z02),CRC/TRR167(Project ID 259373024,Z01),CRC1453(Project ID 431984000,S01),TRR 359(Project ID 491676693,Z01),and FOR 5476 UcarE(Project ID 493802833,P07)funding from the German Federal Ministry of Education and Research(BMBF)within the Medical Informatics Funding SchemePM4Onco-FKZ 01ZZ2322A.G.A.is supported by the BMBF via EkoEstMed-FKZ 01ZZ2015.O.Gr.is supported by the DFG,within CRC1160(Project ID 256073931,C02),CRC/TRR167(Project ID 259373024,A08),CRC1425(Project ID 422681845,P10),GRK2606(Project ID 423813989)the European Research Council(ERC)through Starting Grant 337689,Proof-of-Concept Grant 966687the EU-H2020-MSCA-COFUND EURIdoc program(No.101034170)L.H.and O.Gr.are supported by the DFG under Germany’s Excellence Strategy(CIBSS-EXC-2189-Project ID 390939984)the Core Facility AMIRCF(DFGRIsources N°RI_00052)for support in MR imagingthe Lighthouse Core Facility of the Medical Faculty,University of Freiburg(Project Numbers 2023/A2-Fol,2021/B3-Fol)the DKTK,and the DFG(Project Number 450392965)The Proteomic Platform-Core Facility was supported by the Medical Faculty of the University of Freiburg to O.S.(2021/A3-Sch2023/A3-Sch).
文摘Oncogenic KRAS mutations drive metabolic reprogramming in pancreatic ductal adenocarcinoma(PDAC).Src-homology 2 domaincontaining phosphatase 2(SHP2)is essential for full KRAS activity,and promising dual SHP2/mitogen-activated protein kinase(MAPK)inhibition is currently being tested in clinical trials.Exploitable metabolic adaptations may contribute to invariably evolving resistance.To understand the metabolic changes induced by dual inhibition,we comprehensively tested human and murine PDAC cell lines,endogenous tumor models,and patient-derived organoids,which are representative of the full spectrum of PDAC molecular subtypes.We found that dual SHP2/mitogen-activated protein kinase kinase(MEK1/2)inhibition induces major alterations in mitochondrial mass and function,impacts reactive oxygen species(ROS)homeostasis and triggers lipid peroxidase dependency.Anabolic pathways,autophagy and glycolysis were also profoundly altered.However,most strikingly,mitochondrial remodeling was evident,persisting into a therapy-resistant state.The resulting vulnerability to the induction of ferroptotic cell death via the combination of vertical SHP2/MEK1/2 with glutathione peroxidase(GPX4)inhibition was largely independent of the PDAC molecular subtype and was confirmed with direct targeting of RAS.The triple combination of SHP2/MEK1/2 inhibition and the ferroptosis-inducing natural compound withaferin A suppressed tumor progression in an endogenous PDAC tumor model in vivo.Our study offers a metabolic leverage point to reinforce RAS pathway interference for targeted PDAC treatment.
文摘Objective:To characterize the tumor-suppressive role of LINC00936 in non-small cell lung cancer(NSCLC)through mechanistic exploration of its regulatory pathways.Methods:Bioinformatics interrogation of TCGA/NSCLC cohorts assessed LINC00936 expression,clinical correlations,and immune contexture.Functional enrichment analyses predicted pathway associations.In H1299 cells,LINC00936 overexpression(plasmid)and knockdown(siRNA)models were validated by RT-qPCR.Transcriptomic profiling identified differentially expressed genes(DEGs)subjected to KEGG pathway analysis.Results:LINC00936 was significantly downregulated in NSCLC tissues(TCGA,P<0.05)and cell lines(vs.16-HBE,P<0.05),correlating with poor prognosis and altered tumor-infiltrating immune subsets.DEG enrichment implicated Ras/MAPK signaling as the dominant pathway(FDR<0.05).Successful LINC00936 modulation(overexpression/knockdown,P<0.05)confirmed its regulatory capacity.Conclusion:LINC00936 acts as a tumor suppressor in NSCLC via Ras/MAPK pathway modulation,proposing its therapeutic candidacy for precision oncology strategies.
基金supported by the Natural Science Foundation of Hebei Province(H2021206037)the Government-funded Project on Training of Outstanding Clinical Medical Personnel of Hebei Province in the year 2021(303-16-20-06)the Medical Research Project of Hebei Provincial Health Commission(20230031).
文摘Background:Glioma is the most common tumor of the central nervous system with a poor prognosis.This study aims to explore the role of calcium/calmodulin-dependent protein kinase IIβ(CAMK2B)in regulating the malignant progression of glioma cells,as well as the molecular mechanisms underlying these malignant behaviors.Methods:The correlation between CAMK2B expression in gliomas and patient prognosis was analyzed using immunohistochemistry,quantitative reverse transcription polymerase chain reaction(qRT-PCR),and western blot.Furthermore,the study explored the role of CAMK2B in glioma cell proliferation,invasion,and migration using cell counting kit-8(CCK-8),5-Ethynyl-2′-deoxyuridine(EdU),wound healing,transwell,and in vivo tumor xenograft assays.Result:Patients with high CAMK2B expression exhibited significantly better prognostic outcomes compared to those with low expression levels.Furthermore,CAMK2B expression was significantly lower in glioma tissues and cells compared to both normal brain tissue and human astrocyte cell lines.Notably,overexpression of CAMK2B in glioma cells led to an approximate 40%reduction in proliferative capacity and a 60–70%decrease in invasive and migratory abilities,compared to control glioma cells.These differences were statistically significant at p<0.05.Conversely,knockdown of CAMK2B using siRNA-CAMK2B significantly enhanced the proliferative,invasive,and migratory capabilities of glioma cells in both in vitro and in vivo settings,enhancing these abilities by 1.5 to 3 times.Notably,these effects were reversed through the application of the Rat Sarcoma viral oncogene homolog(Ras)pathway inhibitor,Salirasib.Western blot analysis revealed that knockdown of CAMK2B led to activation of the Ras/Rapidly Accelerated Fibrosarcoma(Raf)/Mitogen-activated protein kinase kinase(MEK)/Extracellular signal-regulated kinase(ERK)signaling pathway in glioma cell lines,whereas overexpression of CAMK2B resulted in the suppression of this pathway.Conclusion:CAMK2B inhibits glioma proliferation,invasion,andmigration through the Ras/Raf/MEK/ERK signaling pathway.
基金the Independent Subject of Guangxi Key Laboratory of Medicinal Resources Protection and Genetic Improvement(No.KL2022ZZ01)Guangxi Innovation-Driven Development Project(No.GuiKe AA18242040)+3 种基金the National Key R&D Program of China(Nos.2019YFC1711000 and 2019YFC1711008)the Innovation Team Project of Guangxi Botanical Garden of Medicinal Plant(No.GuiYaoChuang2019002)the Innovation and Demonstration of Guangxi Academician Workstation(No.2021AV07007)Government Guided Local Science and Technology Development Project(No.GuiKeZY 20198018).
文摘Nasopharyngeal carcinoma(NPC)is the third most common malignancy with a high recurrence and metastasis rate in South China.Natural compounds extracted from traditional Chinese herbal medicines have been developed and utilized for the treatment of a variety of cancers with modest properties and slight side effects.Maackiain(MA)is a type of flavonoid that was first isolated from leguminous plants,and it has been reported to relieve various nervous system disorders and exert anti-allergic as well as antiinflammatory effects.In this study,we demonstrated that MA inhibited proliferation,arrested cell cycle and induced apoptosis in nasopharyngeal carcinoma CNE1 and CNE2 cells in vitro and in vivo.The expression of the related proteins associated with these processes were consistent with the above effects.Moreover,transcriptome sequencing and subsequent Western blot experiments revealed that inhibition of the MAPK/Ras pathway may be responsible to the anti-tumor effect of MA on NPC cells.Therefore,the effects of MA and an activator of this pathway,tertiary butylhydroquinone(TBHQ),alone or combination,were investigated.The results showed TBHQ neutralized the inhibitory effects of MA.These data suggest that MA exerts its anti-tumor effect by inhibiting the MAPK/Ras signaling pathway and it has the potential to become a treatment for patients with NPC.
基金We thank for the funding support from the National Natural Science Foundation of China(No.81874408,No.81973914 and No.81573721)the Natural Science Foundation of Hunan Province,China(No.2019JJ40216)Study Fundation of Hunan Provincial Education Department(No.19B439).
文摘Objective To assess the effects of Yi Qi Jie Du Formula(YQJDF)combined with salinomycin(SAL)on nasopharyngeal carcinoma stem cells(NPC-SCs)and investigate the underlying molecular mechanisms.Methods Cell counting methods,the CCK-8 assay,transwell migration assay and JC-1 staining,were used to observe the effects of the combination on the proliferation,migration and apoptosis of NPC stem cells,respectively.Western blot was used to detect the levels of protein in NPC-SCs.Results YQJDF combined with SAL had a synergistic effect on the inhibition of proliferation and migration and induction of NPCSC apoptosis.Mechanistically,YQJDF combined with SAL synergistically upregulated the levels of apoptotic proteins,including cleaved Caspase-3,cleaved Caspase-7 and cleaved Caspase-9.Moreover,YQJDF combined with SAL synergistically decreased the levels of CD44,p-c-Src,Ras,p-PKCδ,p-MEK,p-c-Raf,p-ERK1/2 and p-AKT proteins.Conclusions The combination of YQJDF and SAL has a synergistic effect on the inhibition of NPC-SC proliferation and migration and induction of apoptosis,which may be closely related to the downregulation of the CD44/Ras signaling pathway.
基金funded by the National Natural Science Foundation for Young Scholars of China,No.81101362 and 81401784the Key Project of National Natural Science Foundation of China,No.81330042
文摘An increasing number of studies report that the Ras/Raf/extracellular signal-regulated kinase 1/2 （ERK1/2） signaling pathway has a death-promoting apoptotic function in neural cells. We hypothesized that the Ras/Raf/ERK1/2 signaling pathway may be abnormally regulated in rat injured spinal cord models. The weight drop method was used to establish rat spinal cord injury at T9. Western blot analysis and immunohistochemical staining revealed Ras expression was dramatically elevated, and the phosphorylations of A-Raf, B-Raf and C-Raf were all upregulated in the injured spinal cord. Both mitogen-activated protein kinase kinase 1/2 and ERK1/2, which belong to the Ras/Raf signaling kinases, were upregulated. These results indicate that Ras/Raf/ ERK1/2 signaling may be upregulated in injured spinal cord and are involved in recovery after spinal cord injury.
基金National Natural Science Foundation of China(No.81574017)。
文摘Objective:To investigate the effect of Bushen Antai Granule on the mRNA and protein expression of Ras protein/mitogen activated protein kinase mediated by vascular endothelial growth factor receptor 2 with small interference RNA interference.Methods:Method to construct the placenta microvascular endothelial cells,and the preparation of kidney fetus granule drug-containing serum,select the best drug-containing serum concentration,it can be divided into normal group,the serum siRNA-NC normal serum group,drug serum,siRNA normal serum group,siRNA drug serum group,using real-time fluorescent quantitative PCR,Western blotting,immunofluorescence test respectively the RAS/MAPK mRNA and protein expression.Results:Results there was no significant difference in Ras and MAPK mRNA and protein expression between the normal group and the negative control group(P>0.05).The mRNA and protein expressions of Ras and MAPK in the drug serum group were significantly higher than those in the normal serum group(P<0.01).Ras and MAPK mRNA and protein expression were significantly decreased in siRNA1 normal serum group compared with normal serum group(P<0.01).Ras,MAPK mRNA and protein expression in siRNA1 drug serum group were significantly different from that in siRNA1 normal serum group(P<0.01).Conclusion:Conclusion The therapeutic effect of Bushen Antai Granule on recurrent abortion may be realized by upregulation of RAS/MAPK mRNA and protein expression.
基金Liaoning Natural Science Foundation(20180530033)Open Fund of Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications,Liaoning University of Traditional Chinese Medicine。
文摘[Objectives]To explore the neuroprotective effects and mechanism of Longan Aril(LA)effective parts on PC12 cells injured by H2O2.[Methods]The neuroprotective effects of LA were evaluated by the cell viability,SOD and MDA content,apoptosis assay and relative protein expression of Aβand p-Tau.The neuroprotective mechanism of LA was studied by using metabolomics and network pharmacology,and the expressions of RAS/MEK/ERK signaling pathway-related proteins were detected by western blotting.[Results]LA could improve the cell survival rate and SOD content,and reduce apoptosis and expression of Aβand p-tau.Inhibition of RAS/MEK/ERK signaling pathway is a possible mechanism of LA neuroprotective effects.[Conclusions]LA has a neuroprotective effects in vitro and be likely to inhibit the process of AD by inhibition of RAS/MEK/ERK signalling pathway.
基金supported by grants from the National Key Research and Development Program of China(2017YFA0105002,Y.Z.2017YFA0104402,L.L.)Joint Funds of the National Natural Science Foundation of China(U1738111,Y.Z.)+1 种基金the China Manned Space Flight Technology Project(TZ-1)the National Natural Science Foundation Youth Fund(31800741,L.S.).
文摘Spaceflight-associated immune system weakening ultimately limits the ability of humans to expand their presence beyond the earth's orbit. A mechanistic study of microgravity-regulated immune cell function is necessary to overcome this challenge. Here, we demonstrate that both spaceflight (real) and simulated microgravity significantly reduce macrophage differentiation, decrease macrophage quantity and functional polarization, and lead to metabolic reprogramming, as demonstrated by changes in gene expression profiles. Moreover, we identified RAS/ERK/NFκB as a major microgravity-regulated pathway. Exogenous ERK and NFκB activators significantly counteracted the effect of microgravity on macrophage differentiation. In addition, microgravity also affects the p53 pathway, which we verified by RT-qPCR and Western blot. Collectively, our data reveal a new mechanism for the effects of microgravity on macrophage development and provide potential molecular targets for the prevention or treatment of macrophage differentiation deficiency in spaceflight.
基金supported by the National Natural Science Foundation of China,Nos.81401002 (to SSZ),81801 053 (to XQZ)。
文摘Stromal cell-derived factor-1 and its receptor C-X-C chemokine receptor 4(CXCR4) have been shown to regulate neural regeneration after stroke.Howeve r,whether stromal cell-derived factor-1 receptor CXCR7,which is widely distributed in the develo ping and adult central nervous system,participates in neural regeneration remains poorly unde rstood.In this study,we established rat models of focal cerebral ischemia by injecting endothelin-1 into the cerebral co rtex and striatum.Starting on day 7 after injury,CXCR7-neutralizing antibody was injected into the lateral ventricle using a micro drug delivery system for 6 consecutive days.Our results showed that CXCR7-neutralizing antibody increased the total length and number of sprouting co rticospinal tra ct fibers in rats with cerebral ischemia,increased the expression of vesicular glutamate transporter 1 and growth-related protein 43,marke rs of the denervated spinal cord synapses,and promoted the differentiation and maturation of oligodendrocyte progenitor cells in the striatum.In addition,CXCR7 antibody increased the expression of CXCR4 in the striatum,increased the protein expression of RAS and ERK1/2 associated with the RAS/ERK signaling pathway,and im proved rat motor function.These findings suggest that CXCR7 improved neural functional recovery after ischemic stroke by promoting axonal regeneration,synaptogenesis,and myelin regeneration,which may be achieved by activation of CXCR4 and the RAS/ERK1/2 signaling pathway.
文摘Abnormal activation of the Ras/Raf/Mek/Erk signaling cascade plays an important role in glioma. Inhibition of this aberrant activity could effectively hinder glioma cell proliferation and promote cell apoptosis. To investigate the mechanism of gJioblastoma treatment by neural stem ceiJ trans- plantation with respect to the Ras/Raf/Mek/Erk pathway, C6 glioma cells were prepared in sus- pension and then infused into the rat brain to establish a glioblastoma model. Neural stem cells isolated from fetal rats were then injected into the brain of this glioblastoma model. Results showed that Raf-1, Erk and Bcl-2 protein expression significantly increased, while Caspase-3 protein expression decreased. After transplantation of neural stem cells, Raf-1, Erk and Bcl-2 protein expression significantly decreased, while Caspase-3 protein expression significantly in-creased. Our findings indicate that transplantation of neural stem cells may promote apoptosis of glioma cells by inhibiting Ras/Raf/Mek/Erk signaling, and thus may represent a novel treatment approach for glioblastoma.
基金the National Natural Science Foundation of China,No.50677022
文摘BACKGROUND： Electromagnetic radiation can influence dopamine （DA） synthesis in brain tissues or ceils, but electromagnetic frequencies, intensities, and radiation time can produce different effects. In addition, the signal pathway by which electromagnetic radiation influences DA synthesis remains controversial. OBJECTIVE： To determine tyrosine hydroxylase （TH） expression in PC12 cells and DA levels in cell culture media after different periods of low-frequency pulsed electric field （LF-PEF） stimulation, and to determine how LF-PEF signaling stimulates TH synthesis using inhibitors. DESIGN, TIME AND SETTING： A parallel, controlled, cell experiment was performed at the Laboratory of Cell Biology, School of Life Science, East China Normal University, between January and October 2006. MATERIALS： PC12 cells were purchased from the Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, China. Nerve growth factor was purchased from PeproTech, USA. The protein kinase A inhibitor, H-89, and mitogen-activated protein kinase kinase inhibitor, U0126, were purchased from Sigma, USA. METHODS： （1） Following routine culture in Dulbecco＇s modified eagle medium, primary PC12 cells were stimulated under LF-PEF （pulse frequency 50.Hz, pulse width 20 μs, peak field strength 1 V/m） for 5, 10, 15, 20, and 30 minutes. （2） Inhibitors （H-89 or U0126, 1 μmol/L） were added 30 minutes before LF-PEF stimulation for 10 minutes. MAIN OUTCOME MEASURES： （1） TH expression was determined by Western blot in PC12 cells at 0.5, 1,2, 3, and 4 days after LF-PEF stimulation. Similarly, DA was measured by high-performance liquid chromatography in media at 2, 3, 4, or 5 days after LF-PEE （2） TH expression was detected 1 day after H-89 or U0126 treatment and LF-PEE RESULTS： （1） Short-term LF-PEF stimulation （5 and 10 minutes） increased TH expression and media DA levels after short-term culture （2 days） （P 〈 0.01）, but both parameters decreased with longer culture （3 4 days） （P 〈 0.01）. Long-term LF-PEF stimulation （15, 20, or 30 minutes） decreased TH and DA synthesis, followed by a rapid increase （P 〈 0.01）. （2） H89 could completely inhibit TH expression in PC12 cells stimulated by LF-PEF for 10 minutes, while the inhibition rate of U0126 was 53.2%. CONCLUSION： Short-term LF-PEF first promotes then inhibits, while long-term LF-PEF first inhibits then promotes, TH and DA synthesis. LF-PEF stimulation regulates TH expression primarily by activating protein kinase A to regulate DA synthesis.
文摘Subject Code:H16With the support by the National Natural Science Foundation of China,a collaborative study by the research groups led by Prof.Xiao Zhixiong(肖智雄)from the College of Life Science,Sichuan University demonstrates thatΔNp63αis a common inhibitory target in oncogenic PI3K/Ras/Her2-induced
基金supported by the National Key Research and Development Project(2019YFC1804504 and 2019YFC1804503)National Natural Science Foundation of China(41731279)Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program(2017BT01Z032).
文摘Tetrabromobisphenol A(TBBPA)is a flame retardant that adversely affects the environment and human health.The present study exposed HepG2 cells to low concentrations of TBBPA daily to investigate the changes in gene regulation,mainly related to pathways associated with the endocrine system.The quantitative polymerase chain reaction(qPCR)confirmed that prolonged exposure gradually activated the thyroid hormone and parathyroid hormone signaling pathways.The expression levels of genes related to the thyroid hormone signaling pathway were upregulated(1.15-8.54 times)after five generations of exposure to 1 and 81 nM TBBPA.Furthermore,co-exposure to 81 nM TBBPA and 0.5 nM thyroid hormone receptor antagonist for five generations significantly reduced the expression of thyroid hormone and parathyroid hormone receptors.Meanwhile,81 nM TBBPA inhibited the activation of the Ras pathway and downregulated Ras gene expression level(3.7 times),indicating the association between the toxic effect and thyroid hormone receptors.Additionally,our experiments revealed that the thyroid hormone pathway regulated the induction of the Ras signaling pathway by TBBPA.The study thus proves that daily exposure to TBBPA interferes with the thyroid hormone signaling pathway and subsequently the endocrine system.
基金funded by the National Key R&D Program of China(2022YFF1000304)the National Natural Sciences Foundation of China(32273102 and 31972782)+1 种基金the Strategic Priority Research Program of the Chinese Academy of Sciences(Grant No.XDA24030105)the Taishan Scholars Program.
文摘Growth of the Pacific white shrimp Litopenaeus vannamei,the most important farmed crustacean,has consistently been a focal point for breeders.Over the past decades,some candidate genes for shrimp growth have been identified.However,fur-ther research is needed to elucidate the molecular regulatory mechanism of these genes.LvMmd2 was previously identified as a candidate gene that may inhibit the growth of L.vannamei.In this study,we analyzed the genotype and expression of the LvMmd2 gene in a breeding family and indicated its role as a growth-inhibiting gene.We found that LvMmd2 co-localized with its homolog LvPAQR3 at the Golgi apparatus.Using co-immunoprecipitation(Co-IP)and DUAL membrane system yeast two-hybrid(MbY2H),we indicated the interactions between LvMmd2 and LvPAQR3,LvPAQR3 and LvRaf1,as well as LvMmd2 and LvRho.These results suggest that LvMmd2 directly and indirectly regulates the Ras signaling pathway.Furthermore,we show that the LvMmd2 gene may indirectly affect the PI3K/AKT,insulin,and Hippo signaling pathways to regulate cell proliferation and differentiation via LvPAQR3 and LvRaf1.Through transcriptome and MbY2H analyses,we have also revealed the interaction between LvMmd2 and proteins involved in growth,immunity,protein transport,synthesis,and modification.These findings demonstrate the various molecular pathways through which LvMmd2 regulates L.vannamei growth.This study provides insights into the mechanism of shrimp growth regulated by Mmd2,enhances our understanding of LvMmd2 function,and highlights its potential application in shrimp breeding.
基金supported by grants from the National Natural Science Foundation of China(Grant Nos.32170594 and 31870809)the Province and Ministry Coconstruction Major Program of Medical Science and Technique Foundation of Henan Province(Grant No.SBGJ202001007)the Special Fund for Young and Middle School Leaders of Henan Health Commission(Grant No.HNSWJW-2020017),China.
文摘RNA modifications affect many biological processes and physiological diseases.The 5-methylcytosine(m^(5)C)modification regulates the progression of multiple tumors.However,its characteristics and functions in hepatocellular carcinoma(HCC)remain largely unknown.Here,we found that HCC tissues had a higher m^(5)C methylation level than the adjacent normal tissues.Transcriptome analysis revealed that the hypermethylated genes mainly participated in the phosphokinase signaling pathways,such as the Ras and PI3K-Akt pathways.The m^(5)C methyltransferase NSUN2 was highly expressed in HCC tissues.Interestingly,the expression of many genes was positively correlated with the expression of NSUN2,including GRB2,RNF115,AATF,ADAM15,RTN3,and HDGF.Real-time PCR assays further revealed that the expression of the mRNAs of GRB2,RNF115,and AATF decreased significantly with the down-regulation of NSUN2 expression in HCC cells.Furthermore,NSUN2 could regulate the cellular sensitivity of HCC cells to sorafenib via modulating the Ras signaling pathway.Moreover,knocking down NSUN2 caused cell cycle arrest.Taken together,our study demonstrates the vital role of NSUN2 in the progression of HCC.
