Cellular communication network factor 2(CCN2)is a secreted extracellular matrix-associated protein,and its aberrantly increased expression has been implicated in a diversity of diseases involving pathological processe...Cellular communication network factor 2(CCN2)is a secreted extracellular matrix-associated protein,and its aberrantly increased expression has been implicated in a diversity of diseases involving pathological processes of fibrosis,chronic inflammation,or tissue injury,which has promoted the evaluation of CCN2 as therapeutic targets for multiple disorders.However,human phenotypes associated with CCN2 deficiency have remained enigmatic;variants in CCN2 have not yet been associated with a human phenotype.Here,we collected families diagnosed with spondyloepimetaphyseal dysplasia(SEMD),and screened candidate pathogenic genes for families without known genetic causes using next-generation sequencing.We identified a monoallelic variant in signal peptide of CCN2(NM_001901.2:c.65 G>C[p.Arg22Pro])as the cause of SEMD in 14 subjects presenting with different degree of short stature,premature osteoarthritis,and osteoporosis.Affected subjects showed decreased serum CCN2 levels.Cell lines harboring the variant displayed decreased amount of CCN2 proteins in culture medium and an increased intracellular retention,indicating impaired protein secretion.And the variant weakened the stimulation effect of CCN2 on osteogenesis of bone marrow mesenchymal stem cells.Zebrafish ccn2a knockout model and osteoblast lineage-specific Ccn2-deficient mice(Ccn2fl/fl;Prx1Cre)partially recapitulated the phenotypes including low bone mass observed in affected subjects.Pathological mechanism implicated in the skeletal abnormality in Ccn2fl/fl;Prx1Cre mice involved decreased bone formation,increased bone resorption,and abnormal growth plate formation.Collectively,our study indicate that monoallelic variants in CCN2 lead to a human inherited skeletal dysplasia,and highlight the critical role of CCN2 in osteogenesis in human.展开更多
目的探讨血管内皮生长因子(VEGF)诱导的成骨细胞中结缔组织生长因子(CTGF/CCN2)对人脐静脉血管内皮细胞(HUVECs)的影响。方法用Real time PCR法及ELISA法检测VEGF诱导成骨细胞(OSE)中CCN2含量;制备成骨细胞(OSE)上清液;将细胞分为contro...目的探讨血管内皮生长因子(VEGF)诱导的成骨细胞中结缔组织生长因子(CTGF/CCN2)对人脐静脉血管内皮细胞(HUVECs)的影响。方法用Real time PCR法及ELISA法检测VEGF诱导成骨细胞(OSE)中CCN2含量;制备成骨细胞(OSE)上清液;将细胞分为control组、OSE组和VEGF-OSE组(n=3)。用小干扰RNA(siRNA)转染法抑制成骨细胞中CCN2的表达;Transwell法检测内皮细胞迁移;Matrigel实验检测管样结构形成能力。结果VEGF呈时间和剂量依赖性上调成骨细胞中CCN2 mRNA和蛋白的表达;CCN2可促进内皮细胞的迁移和管样结构形成(P<0.05),当CCN2被siRNA基因沉默或者加入CCN2抗体后,CCN2对内皮细胞迁移和管样结构形成的促进作用均受到明显抑制(P<0.05)。结论 VEGF通过上调成骨细胞中CCN2的表达,促内皮细胞(HUVECs)的迁移和血管生成。展开更多
Background:We previously confirmed that mechanical stimulation is an important factor in the repair of tendon-bone insertion(TBI)injuries and that mechanoreceptors such as transient receptor potential ion-channel subf...Background:We previously confirmed that mechanical stimulation is an important factor in the repair of tendon-bone insertion(TBI)injuries and that mechanoreceptors such as transient receptor potential ion-channel subfamily V member 4(TRPV4;also known as transient receptor potential vanilloid 4)are key to transforming mechanical stimulation into intracellular biochemical signals.This study aims to elucidate the mechanism of mechanical stimulation regulating TRPV4.Methods:Immunohistochemical staining and western blotting were used to evaluate cartilage repair at the TBI after injury.The RNA expression and protein expression of mechanoreceptors and key pathway molecules regulating cartilage proliferation were analyzed.TBI samples were collected for transcriptome sequencing to detect gene expression.Calcium-ion imaging and flow cytometry were used to evaluate the function of TPRV4 and cellular communication network factor 2(CCN2)after the administration of siRNA,recombinant adenovirus and agonists.Results:We found that treadmill training improved the quality of TBI healing and enhanced fibrochondrocyte proliferation.The transcriptome sequencing results suggested that the elevated expression of the mechanistically stimulated regulator CCN2 and the exogenous administration of recombinant human CCN2 significantly promoted TRPV4 protein expression and fibrochondrocyte proliferation.In vitro,under mechanical stimulation conditions,small interfering RNA(siRNA)-CCN2 not only inhibited the proliferation of primary fibrochondrocytes but also suppressed TRPV4 protein expression and activity.Subsequently,primary fibrochondrocytes were treated with the TRPV4 agonist GSK1016790A and the recombinant adenovirus TRPV4(Ad-TRPV4),and GSK1016790A partially reversed the inhibitory effect of siRNA-CCN2.The phosphoinositide 3-kinase/protein kinase B(PI3K/AKT)signaling pathway participated in the above process.Conclusions:Mechanical stimulation promoted fibrochondrocyte proliferation and TBI healing by activating TRPV4 channels and the PI3K/AKT signaling pathway,and CCN2 may be a key regulatory protein in maintaining TRPV4 activation.展开更多
[目的]探讨肾细胞癌中TXNDC5的表达及其对786-O细胞增殖、侵袭和迁移的影响。[方法]2020年8月-2023年8月102例肾细胞癌患者纳入研究,免疫组化染色测定TXNDC5表达水平,χ^(2)检验分析TXNDC5表达与肾细胞癌临床病理特征的关系。同时设肾...[目的]探讨肾细胞癌中TXNDC5的表达及其对786-O细胞增殖、侵袭和迁移的影响。[方法]2020年8月-2023年8月102例肾细胞癌患者纳入研究,免疫组化染色测定TXNDC5表达水平,χ^(2)检验分析TXNDC5表达与肾细胞癌临床病理特征的关系。同时设肾细胞癌细胞786-O组、sh-NC组、TXNDC5-inhibitor组、TXNDC5-mimics组,测定各组细胞增殖、侵袭、凋亡以及TXNDC5、CCN2水平。[结果]肾细胞癌组TXNDC5蛋白表达阳性率高于癌旁组(P<0.05)。TXNDC5蛋白表达与肾细胞癌临床病理特征相关。TXNDC5-inhibitor组OD值(0.78±0.05 vs 0.41±0.03)、存活率(78.56%±6.18%vs 42.11%±7.24%)、单克隆形成数目[(619.33±89.20)/个]、穿膜数[(323.85±84.25 vs 93.85±95.34)/个]、TXNDC5、CCN2 mRNA和蛋白表达水平低于肾细胞癌细胞786-O组、sh-NC组,凋亡率高于肾细胞癌细胞786-O组、sh-NC组(P<0.05);TXNDC5-mimics组OD值、存活率、单克隆形成数目、穿膜数、TXNDC5、CCN2 mRNA和蛋白表达水平高于肾细胞癌细胞786-O组、sh-NC组,凋亡率低于肾细胞癌细胞786-O组、sh-NC组(P<0.05);TXNDC5-mimics组OD值、存活率、单克隆形成数目、穿膜数、TXNDC5、CCN2 mRNA和蛋白表达水平高于TXNDC5-inhibitor组,凋亡率低于TXNDC5-inhibitor组(P<0.05)。[结论]TXNDC5下调可以抑制肾细胞癌细胞的增殖、迁移和侵袭,同时诱导细胞凋亡,这一过程与调节CCN2相关。展开更多
Osteosarcoma (OS) is the most common histological form of primary bone cancer in childhood cancer and young adults. At present, OS is widely investigated because of the interaction between the tumor and bone microenvi...Osteosarcoma (OS) is the most common histological form of primary bone cancer in childhood cancer and young adults. At present, OS is widely investigated because of the interaction between the tumor and bone microenvironment and the effect of such interaction on OS progression and metastasis.1 The connective tissue growth factor (CTGF), also known as cellular communication network factor 2 (CCN2), is a secreted extracellular matrix-associated protein. CTGF is as active as the regulators of signaling activities of several different pathways and an orchestrator of their cross-talk.2 Therefore, we conducted experiments to investigate the effects of CTGF on OS tumor progress and the cross-talk with stromal cells in the tumor microenvironment.展开更多
基金funded by the National Key Research and Development Program of China (2018YFA0800801)the National Basic Research Program of China (2014CB942903)+1 种基金the National Natural Science Foundation of China (81900807)Shanghai Research Center for Endocrine and Metabolic Diseases (2022ZZ01002)。
文摘Cellular communication network factor 2(CCN2)is a secreted extracellular matrix-associated protein,and its aberrantly increased expression has been implicated in a diversity of diseases involving pathological processes of fibrosis,chronic inflammation,or tissue injury,which has promoted the evaluation of CCN2 as therapeutic targets for multiple disorders.However,human phenotypes associated with CCN2 deficiency have remained enigmatic;variants in CCN2 have not yet been associated with a human phenotype.Here,we collected families diagnosed with spondyloepimetaphyseal dysplasia(SEMD),and screened candidate pathogenic genes for families without known genetic causes using next-generation sequencing.We identified a monoallelic variant in signal peptide of CCN2(NM_001901.2:c.65 G>C[p.Arg22Pro])as the cause of SEMD in 14 subjects presenting with different degree of short stature,premature osteoarthritis,and osteoporosis.Affected subjects showed decreased serum CCN2 levels.Cell lines harboring the variant displayed decreased amount of CCN2 proteins in culture medium and an increased intracellular retention,indicating impaired protein secretion.And the variant weakened the stimulation effect of CCN2 on osteogenesis of bone marrow mesenchymal stem cells.Zebrafish ccn2a knockout model and osteoblast lineage-specific Ccn2-deficient mice(Ccn2fl/fl;Prx1Cre)partially recapitulated the phenotypes including low bone mass observed in affected subjects.Pathological mechanism implicated in the skeletal abnormality in Ccn2fl/fl;Prx1Cre mice involved decreased bone formation,increased bone resorption,and abnormal growth plate formation.Collectively,our study indicate that monoallelic variants in CCN2 lead to a human inherited skeletal dysplasia,and highlight the critical role of CCN2 in osteogenesis in human.
文摘目的探讨血管内皮生长因子(VEGF)诱导的成骨细胞中结缔组织生长因子(CTGF/CCN2)对人脐静脉血管内皮细胞(HUVECs)的影响。方法用Real time PCR法及ELISA法检测VEGF诱导成骨细胞(OSE)中CCN2含量;制备成骨细胞(OSE)上清液;将细胞分为control组、OSE组和VEGF-OSE组(n=3)。用小干扰RNA(siRNA)转染法抑制成骨细胞中CCN2的表达;Transwell法检测内皮细胞迁移;Matrigel实验检测管样结构形成能力。结果VEGF呈时间和剂量依赖性上调成骨细胞中CCN2 mRNA和蛋白的表达;CCN2可促进内皮细胞的迁移和管样结构形成(P<0.05),当CCN2被siRNA基因沉默或者加入CCN2抗体后,CCN2对内皮细胞迁移和管样结构形成的促进作用均受到明显抑制(P<0.05)。结论 VEGF通过上调成骨细胞中CCN2的表达,促内皮细胞(HUVECs)的迁移和血管生成。
基金supported by the National Natural Science Foundation of China(NSFC,Nos.82130071 and 82072516).
文摘Background:We previously confirmed that mechanical stimulation is an important factor in the repair of tendon-bone insertion(TBI)injuries and that mechanoreceptors such as transient receptor potential ion-channel subfamily V member 4(TRPV4;also known as transient receptor potential vanilloid 4)are key to transforming mechanical stimulation into intracellular biochemical signals.This study aims to elucidate the mechanism of mechanical stimulation regulating TRPV4.Methods:Immunohistochemical staining and western blotting were used to evaluate cartilage repair at the TBI after injury.The RNA expression and protein expression of mechanoreceptors and key pathway molecules regulating cartilage proliferation were analyzed.TBI samples were collected for transcriptome sequencing to detect gene expression.Calcium-ion imaging and flow cytometry were used to evaluate the function of TPRV4 and cellular communication network factor 2(CCN2)after the administration of siRNA,recombinant adenovirus and agonists.Results:We found that treadmill training improved the quality of TBI healing and enhanced fibrochondrocyte proliferation.The transcriptome sequencing results suggested that the elevated expression of the mechanistically stimulated regulator CCN2 and the exogenous administration of recombinant human CCN2 significantly promoted TRPV4 protein expression and fibrochondrocyte proliferation.In vitro,under mechanical stimulation conditions,small interfering RNA(siRNA)-CCN2 not only inhibited the proliferation of primary fibrochondrocytes but also suppressed TRPV4 protein expression and activity.Subsequently,primary fibrochondrocytes were treated with the TRPV4 agonist GSK1016790A and the recombinant adenovirus TRPV4(Ad-TRPV4),and GSK1016790A partially reversed the inhibitory effect of siRNA-CCN2.The phosphoinositide 3-kinase/protein kinase B(PI3K/AKT)signaling pathway participated in the above process.Conclusions:Mechanical stimulation promoted fibrochondrocyte proliferation and TBI healing by activating TRPV4 channels and the PI3K/AKT signaling pathway,and CCN2 may be a key regulatory protein in maintaining TRPV4 activation.
文摘[目的]探讨肾细胞癌中TXNDC5的表达及其对786-O细胞增殖、侵袭和迁移的影响。[方法]2020年8月-2023年8月102例肾细胞癌患者纳入研究,免疫组化染色测定TXNDC5表达水平,χ^(2)检验分析TXNDC5表达与肾细胞癌临床病理特征的关系。同时设肾细胞癌细胞786-O组、sh-NC组、TXNDC5-inhibitor组、TXNDC5-mimics组,测定各组细胞增殖、侵袭、凋亡以及TXNDC5、CCN2水平。[结果]肾细胞癌组TXNDC5蛋白表达阳性率高于癌旁组(P<0.05)。TXNDC5蛋白表达与肾细胞癌临床病理特征相关。TXNDC5-inhibitor组OD值(0.78±0.05 vs 0.41±0.03)、存活率(78.56%±6.18%vs 42.11%±7.24%)、单克隆形成数目[(619.33±89.20)/个]、穿膜数[(323.85±84.25 vs 93.85±95.34)/个]、TXNDC5、CCN2 mRNA和蛋白表达水平低于肾细胞癌细胞786-O组、sh-NC组,凋亡率高于肾细胞癌细胞786-O组、sh-NC组(P<0.05);TXNDC5-mimics组OD值、存活率、单克隆形成数目、穿膜数、TXNDC5、CCN2 mRNA和蛋白表达水平高于肾细胞癌细胞786-O组、sh-NC组,凋亡率低于肾细胞癌细胞786-O组、sh-NC组(P<0.05);TXNDC5-mimics组OD值、存活率、单克隆形成数目、穿膜数、TXNDC5、CCN2 mRNA和蛋白表达水平高于TXNDC5-inhibitor组,凋亡率低于TXNDC5-inhibitor组(P<0.05)。[结论]TXNDC5下调可以抑制肾细胞癌细胞的增殖、迁移和侵袭,同时诱导细胞凋亡,这一过程与调节CCN2相关。
基金supported by the National Natural Science Foundation of China(No.81102035,and 82060388)Doctor Foundation of Guizhou Provincial People's Hospital(No.GZSYBS[2019]08)+2 种基金Guizhou High-level Innovative Talents Project(No.QKPT[2017]5724-6)Guizhou Department and Platform Talents(No.[2017]5735-31)Science and Technology Department of Guizhou Province(No.QKHJC[2021-396]).
文摘Osteosarcoma (OS) is the most common histological form of primary bone cancer in childhood cancer and young adults. At present, OS is widely investigated because of the interaction between the tumor and bone microenvironment and the effect of such interaction on OS progression and metastasis.1 The connective tissue growth factor (CTGF), also known as cellular communication network factor 2 (CCN2), is a secreted extracellular matrix-associated protein. CTGF is as active as the regulators of signaling activities of several different pathways and an orchestrator of their cross-talk.2 Therefore, we conducted experiments to investigate the effects of CTGF on OS tumor progress and the cross-talk with stromal cells in the tumor microenvironment.
