Objective To investigate effect of tumor necrosis factor-or （TNF-α） on the Src-suppressed C kinase substrate （SSeCKS） in C6 glioma cells. Methods Cultured C6 glioma cells were randomly divided into two groups. In...Objective To investigate effect of tumor necrosis factor-or （TNF-α） on the Src-suppressed C kinase substrate （SSeCKS） in C6 glioma cells. Methods Cultured C6 glioma cells were randomly divided into two groups. In time-dependent group, cells were cultured with TNF-α （2 ng/mL） for 0 h, 1 h, 3 h, 6 h, 12 or 24 h, respectively; in dose-dependent group, cells were cultured with TNF-α （0 ng/mL, 0.02 ng/mL, 0.2 ng/mL, or 2 ng/mL） for 6 h. The expression of SSeCKS was detected by Realtime PCR and Western blot analysis, and immunocytochemistry was used to investigate SSeCKS＇s subcellular localization. Results TNF-α induced rapid phosphorylations of protein kinase C （PKC） substrates in C6 glioma cells, and upregulated SSeCKS expression in a time and concentration dependent manner. Immunocytochemistry suggested that SSeCKS was localized in the cyroplasm and the leading end of podosomal extensions in control groups, while TNF-α induced translocation of SSeCKS perinuclear. This effect could be partly reversed by PKC inhibitor Ro-31-8220. Conclusion TNF-α activates PKC and upregulates SSeCKS expression in C6 glioma cells. These effects are associated with PKC activity, suggesting that SSeCKS plays a role in response to glia activation in PKC mediated pathway.展开更多
It is believed that amyloid-βpeptide（Aβ）plays a central role in the pathogenesis of Alzheimer’s disease（AD）.Thus,the process of amyloid precursor protein（APP）cleavage is a key event and has raised much attent...It is believed that amyloid-βpeptide（Aβ）plays a central role in the pathogenesis of Alzheimer’s disease（AD）.Thus,the process of amyloid precursor protein（APP）cleavage is a key event and has raised much attention in the field of AD research.It is proposed that APP,β-andγ-secretases are all located on the lipid raft,and the meeting of them is an indispensable step for Aβgeneration.Endocytosis can lead to clustering of APP,β-andγ-secretases from separate smaller lipid rafts into a larger one.On the other hand,for myristoylated alanine-rich C kinase substrate（MARCKS）,phosphorylation by protein kinase C（PKC）or interaction with Ca2＋can lead to its release from membrane into cytoplasm.This process induces the release of actins and phosphatidylinositol 4,5-bisphosphate（PIP2）,which are important factors for endocytosis.Thus,the present review proposes that MARCKS may be implicated in Aβgeneration,by modulating free PIP 2 level and actin movement,causing endocytosis.展开更多
BACKGROUND Long non-coding RNAs(lncRNAs)with differential expression characteristics have been found to be closely related to the tumorigenesis and development of gastric cancer(GC),but their specific mechanisms and r...BACKGROUND Long non-coding RNAs(lncRNAs)with differential expression characteristics have been found to be closely related to the tumorigenesis and development of gastric cancer(GC),but their specific mechanisms and roles still need to be further elucidated.AIM To investigate the expression of LINC01268 in GC and its mechanism of affecting GC progression.METHODS Real-time quantitative polymerase chain reaction was used to detect the expression of LINC01268 in GC tissues,cell lines and plasma.The Kaplan-Meier method was used to evaluate the value of LINC01268 in the prognostication of GC patients.An receiver operating characteristic curve was constructed to evaluate the value of LINC01268 in the diagnosis of GC.Transwell migration and invasion assays and wound healing assays were used to confirm the effect of LINC01268 on the invasion and migration of GC cells.The regulatory relationship between LINC01268 and myristoylated alanine rich protein kinase C substrate(MARCKS),the PI3K/Akt signaling pathway,and the epithelial-mesenchymal transition(EMT)process in GC was demonstrated by western blot analysis.RESULTS The expression of LINC01268 was increased in GC tissues and cell lines.The expression level of LINC01268 was significantly correlated with lymph node metastasis,TNM stage,and tumor differentiation in patients with GC.Over-expression of LINC01268 indicated a poor prognosis for patients with GC,and it had a certain auxiliary diagnostic value for GC.In vitro functional experiments proved that the abnormal expression of LINC01268 further activated the PI3K/Akt signaling pathway and promoted EMT by targeting and regulating MARCKS and ultimately promoted the invasion and metastasis of GC.CONCLUSION This study elucidates that LINC01268 in GC may be an oncogene that further activates the PI3K/Akt signaling pathway and EMT by targeting and regulating MARCKS,and ultimately promotes the invasion and metastasis of GC.LINC01268 may be a potential effective target for the treatment of GC.展开更多
目的:研究细菌脂多糖(LPS)、肿瘤坏死因子(TNF-α)对牛肺动脉内皮细胞(BPAEC)Src抑制的蛋白激酶C底物(Src-suppressed C Kinase Substrate,SSeCKS)表达的影响和对细胞骨架结构的影响,探讨SSeCKS参与细胞骨架结构改变的可能机制。方法:应...目的:研究细菌脂多糖(LPS)、肿瘤坏死因子(TNF-α)对牛肺动脉内皮细胞(BPAEC)Src抑制的蛋白激酶C底物(Src-suppressed C Kinase Substrate,SSeCKS)表达的影响和对细胞骨架结构的影响,探讨SSeCKS参与细胞骨架结构改变的可能机制。方法:应用LPS、TNF-α刺激体外培养的BPAEC,应用原位杂交、免疫印迹方法检测不同刺激条件下BPAEC中SSeCKS mRNA和蛋白的表达情况;免疫细胞荧光法观察LPS、TNF-α对内皮细胞中SSeCKS与纤维状肌动蛋白(F-actin)定位和结构的影响。结果:静息状态的BPAEC表达极少量的SSeCKS;经LPS刺激后,SSeCKS表达没有明显变化;而TNF-α以浓度和时间依赖的方式诱导内皮细胞SSeCKS表达增加;LPS和TNF-α刺激后,F-actin发生重构,且SSeCKS向核周、细胞膜纤维、板状伪足聚集;蛋白激酶C(PKC)抑制剂Ro-31-8220抑制LPS和TNF-α对内皮细胞F-actin和SSeCKS细胞内定位改变的影响。结论:TNF-α能够诱导内皮细胞SSeCKS表达增加,PKC参与LPS、TNF-α诱导内皮细胞F-actin的重构和SSeCKS重新分布,提示SSeCKS可能与LPS和TNF-α诱导内皮细胞F-actin的重构有关。展开更多
黏液高分泌是慢性气道炎症性疾病的重要病理特征。豆蔻酰化富丙氨酸C激酶底物蛋白(myristoy-lated alanine-rich C kinase substrate,MARCKS)是黏液分泌过程中的关键蛋白。蛋白激酶C或其他激酶、细胞内钙离子浓度、钙调蛋白、纤维型-肌...黏液高分泌是慢性气道炎症性疾病的重要病理特征。豆蔻酰化富丙氨酸C激酶底物蛋白(myristoy-lated alanine-rich C kinase substrate,MARCKS)是黏液分泌过程中的关键蛋白。蛋白激酶C或其他激酶、细胞内钙离子浓度、钙调蛋白、纤维型-肌动蛋白、磷脂酰肌醇4,5二磷酸、热休克蛋白70等均可通过MARCKS的桥梁作用,介导气道分泌颗粒的胞内运动和黏蛋白的出胞释放。展开更多
基金supported by the National Natural Science Foundation of China(No.30300099)the Natural Science Foundation of Jiangsu Province,China(No.BK2003035)the Natural Science Research Program in College and University of Jiangsu Province(No.03KJB180109).
文摘Objective To investigate effect of tumor necrosis factor-or （TNF-α） on the Src-suppressed C kinase substrate （SSeCKS） in C6 glioma cells. Methods Cultured C6 glioma cells were randomly divided into two groups. In time-dependent group, cells were cultured with TNF-α （2 ng/mL） for 0 h, 1 h, 3 h, 6 h, 12 or 24 h, respectively; in dose-dependent group, cells were cultured with TNF-α （0 ng/mL, 0.02 ng/mL, 0.2 ng/mL, or 2 ng/mL） for 6 h. The expression of SSeCKS was detected by Realtime PCR and Western blot analysis, and immunocytochemistry was used to investigate SSeCKS＇s subcellular localization. Results TNF-α induced rapid phosphorylations of protein kinase C （PKC） substrates in C6 glioma cells, and upregulated SSeCKS expression in a time and concentration dependent manner. Immunocytochemistry suggested that SSeCKS was localized in the cyroplasm and the leading end of podosomal extensions in control groups, while TNF-α induced translocation of SSeCKS perinuclear. This effect could be partly reversed by PKC inhibitor Ro-31-8220. Conclusion TNF-α activates PKC and upregulates SSeCKS expression in C6 glioma cells. These effects are associated with PKC activity, suggesting that SSeCKS plays a role in response to glia activation in PKC mediated pathway.
基金supported by the National Natural Science Foundation of China(No.30701137)
文摘It is believed that amyloid-βpeptide（Aβ）plays a central role in the pathogenesis of Alzheimer’s disease（AD）.Thus,the process of amyloid precursor protein（APP）cleavage is a key event and has raised much attention in the field of AD research.It is proposed that APP,β-andγ-secretases are all located on the lipid raft,and the meeting of them is an indispensable step for Aβgeneration.Endocytosis can lead to clustering of APP,β-andγ-secretases from separate smaller lipid rafts into a larger one.On the other hand,for myristoylated alanine-rich C kinase substrate（MARCKS）,phosphorylation by protein kinase C（PKC）or interaction with Ca2＋can lead to its release from membrane into cytoplasm.This process induces the release of actins and phosphatidylinositol 4,5-bisphosphate（PIP2）,which are important factors for endocytosis.Thus,the present review proposes that MARCKS may be implicated in Aβgeneration,by modulating free PIP 2 level and actin movement,causing endocytosis.
基金Supported by the National Natural Science Foundation of China,No.81070378 and 81270561Natural Science Foundation of Sichuan Province,China,No.2022NSFSC0050 and 2023NSFSC1896.
文摘BACKGROUND Long non-coding RNAs(lncRNAs)with differential expression characteristics have been found to be closely related to the tumorigenesis and development of gastric cancer(GC),but their specific mechanisms and roles still need to be further elucidated.AIM To investigate the expression of LINC01268 in GC and its mechanism of affecting GC progression.METHODS Real-time quantitative polymerase chain reaction was used to detect the expression of LINC01268 in GC tissues,cell lines and plasma.The Kaplan-Meier method was used to evaluate the value of LINC01268 in the prognostication of GC patients.An receiver operating characteristic curve was constructed to evaluate the value of LINC01268 in the diagnosis of GC.Transwell migration and invasion assays and wound healing assays were used to confirm the effect of LINC01268 on the invasion and migration of GC cells.The regulatory relationship between LINC01268 and myristoylated alanine rich protein kinase C substrate(MARCKS),the PI3K/Akt signaling pathway,and the epithelial-mesenchymal transition(EMT)process in GC was demonstrated by western blot analysis.RESULTS The expression of LINC01268 was increased in GC tissues and cell lines.The expression level of LINC01268 was significantly correlated with lymph node metastasis,TNM stage,and tumor differentiation in patients with GC.Over-expression of LINC01268 indicated a poor prognosis for patients with GC,and it had a certain auxiliary diagnostic value for GC.In vitro functional experiments proved that the abnormal expression of LINC01268 further activated the PI3K/Akt signaling pathway and promoted EMT by targeting and regulating MARCKS and ultimately promoted the invasion and metastasis of GC.CONCLUSION This study elucidates that LINC01268 in GC may be an oncogene that further activates the PI3K/Akt signaling pathway and EMT by targeting and regulating MARCKS,and ultimately promotes the invasion and metastasis of GC.LINC01268 may be a potential effective target for the treatment of GC.
文摘目的:研究细菌脂多糖(LPS)、肿瘤坏死因子(TNF-α)对牛肺动脉内皮细胞(BPAEC)Src抑制的蛋白激酶C底物(Src-suppressed C Kinase Substrate,SSeCKS)表达的影响和对细胞骨架结构的影响,探讨SSeCKS参与细胞骨架结构改变的可能机制。方法:应用LPS、TNF-α刺激体外培养的BPAEC,应用原位杂交、免疫印迹方法检测不同刺激条件下BPAEC中SSeCKS mRNA和蛋白的表达情况;免疫细胞荧光法观察LPS、TNF-α对内皮细胞中SSeCKS与纤维状肌动蛋白(F-actin)定位和结构的影响。结果:静息状态的BPAEC表达极少量的SSeCKS;经LPS刺激后,SSeCKS表达没有明显变化;而TNF-α以浓度和时间依赖的方式诱导内皮细胞SSeCKS表达增加;LPS和TNF-α刺激后,F-actin发生重构,且SSeCKS向核周、细胞膜纤维、板状伪足聚集;蛋白激酶C(PKC)抑制剂Ro-31-8220抑制LPS和TNF-α对内皮细胞F-actin和SSeCKS细胞内定位改变的影响。结论:TNF-α能够诱导内皮细胞SSeCKS表达增加,PKC参与LPS、TNF-α诱导内皮细胞F-actin的重构和SSeCKS重新分布,提示SSeCKS可能与LPS和TNF-α诱导内皮细胞F-actin的重构有关。
文摘黏液高分泌是慢性气道炎症性疾病的重要病理特征。豆蔻酰化富丙氨酸C激酶底物蛋白(myristoy-lated alanine-rich C kinase substrate,MARCKS)是黏液分泌过程中的关键蛋白。蛋白激酶C或其他激酶、细胞内钙离子浓度、钙调蛋白、纤维型-肌动蛋白、磷脂酰肌醇4,5二磷酸、热休克蛋白70等均可通过MARCKS的桥梁作用,介导气道分泌颗粒的胞内运动和黏蛋白的出胞释放。
