目的研究G3BP1(Ras-GTPase-activating protein SH3 domain binding protein 1)在宫颈癌组织中的表达情况,分析G3BP1的表达水平及其相关临床意义。方法利用公开的GEO宫颈癌数据集分析G3BP1的mRNA水平在癌和癌旁组织的表达差异情况;取30...目的研究G3BP1(Ras-GTPase-activating protein SH3 domain binding protein 1)在宫颈癌组织中的表达情况,分析G3BP1的表达水平及其相关临床意义。方法利用公开的GEO宫颈癌数据集分析G3BP1的mRNA水平在癌和癌旁组织的表达差异情况;取306例宫颈癌组织制作组织芯片,通过免疫组织化学检测癌组织中G3BP1的表达水平,统计分析G3BP1在宫颈癌中与正常宫颈组织和鳞状上皮内病变组织的表达差异情况,分析宫颈癌组织中G3BP1的表达与患者临床病理资料的相关性及对患者生存预后的影响。结果在GSE6791、GSE7803和GSE7410数据集中,我们发现G3BP1的mRNA水平在宫颈癌组织中的表达显著高于癌旁组织。免疫组织化学检测结果显示G3BP1的蛋白表达随着宫颈鳞状上皮内病变和宫颈癌的发生有逐渐增高的趋势。免疫组织化学和χ2检验结果显示G3BP1高表达与年龄、肿瘤大小、肿瘤分化、肌层浸润、阴道浸润、盆腔淋巴结转移、FIGO分期以及不良预后相关。Cox多因素分析显示G3BP1高表达是该批宫颈癌患者的独立危险预后因素。通过与G3BP1的表达水平整合可以显著促进FIGO分期在判断宫颈癌患者预后中的准确性。结论G3BP1高表达可能是宫颈癌患者的独立危险预后因素,并可能参与宫颈癌的发生发展等进程。展开更多
目的:构建GTP酶激活蛋白SH3功能区结合蛋白1(G3BP1)的真核表达载体p EGFP-C3-G3BP1,并观察其在人食管癌EC109细胞中的表达及定位。方法:采用RT-PCR法从EC109细胞中扩增G3BP1 c DNA全长序列,用限制性内切酶HindⅢ和BamHⅠ双酶切PCR产物...目的:构建GTP酶激活蛋白SH3功能区结合蛋白1(G3BP1)的真核表达载体p EGFP-C3-G3BP1,并观察其在人食管癌EC109细胞中的表达及定位。方法:采用RT-PCR法从EC109细胞中扩增G3BP1 c DNA全长序列,用限制性内切酶HindⅢ和BamHⅠ双酶切PCR产物后克隆至pEGFP-C3载体,转化大肠杆菌DH5α后,挑取阳性克隆提取质粒,经双酶切及测序鉴定;将重组质粒用脂质体法转染EC109细胞,荧光定量RT-PCR和Western印迹检测G3BP1的表达,荧光显微镜观察G3BP1的定位。结果:目的基因G3BP1的序列完全正确,并在EC109细胞中获得表达,荧光显微镜观察显示G3BP1定位于细胞质。结论:构建了p EGFP-C3-G3BP1真核表达载体,并在EC109细胞中过表达,G3BP1蛋白定位于细胞质,形成应激颗粒(stress granules,SGs)。为进一步探讨G3BP1在食管癌细胞中的功能及其与SGs的相关性奠定了基础。展开更多
目的:探讨岩藻多糖对胰腺癌的影响,并分析其机制。方法:MTT法分析细胞增殖抑制率,GEPIA数据库分析G3BP1在胰腺癌组织中的表达水平及与生存率的关系。qRT-PCR分析GTP酶激活蛋白结合蛋白1(Ras-GTPase-activating protein binding proteins...目的:探讨岩藻多糖对胰腺癌的影响,并分析其机制。方法:MTT法分析细胞增殖抑制率,GEPIA数据库分析G3BP1在胰腺癌组织中的表达水平及与生存率的关系。qRT-PCR分析GTP酶激活蛋白结合蛋白1(Ras-GTPase-activating protein binding proteins,G3BP1)水平,Western blot法分析p-NF-κBp65、NF-κBp65和IκB-α水平。免疫共沉淀检测G3BP1与p-NF-κBp65之间相互作用。敲低或G3BP1过表达,观察其对岩藻多糖调控细胞增殖以及NF-κB信号通路的影响。裸鼠成瘤实验验证岩藻多糖对裸鼠体内瘤体的瘤重、瘤体积及G3BP1、p-NF-κBp65、NF-κBp65和IκBα水平的影响。结果:1~32μg/mL岩藻多糖抑制capan-1细胞增殖,岩藻多糖48 h IC_(50)为7.729μg/mL。G3BP1在胰腺癌肿瘤组织中的表达明显高于正常组织,G3BP1高表达患者的生存率低于G3BP1低表达患者。2、4、8μg/mL岩藻多糖能下调G3BP1、p-NF-κBp65,上调IκBα水平。Co-IP实验发现G3BP1与p-NF-κBp65相互结合,并且岩藻多糖作用后结合能力降低。敲低G3BP1能促进岩藻多糖抑制capan-1细胞增殖,下调G3BP1、p-NF-κBp65,上调IκBα水平(P<0.05);G3BP过表达能下调岩藻多糖抑制capan-1细胞增殖效果,上调G3BP1、p-NF-κBp65,下调IκBα水平(P<0.05)。体内实验显示,敲低G3BP1能促进岩藻多糖减少瘤体体积、瘤体质量,下调瘤体G3BP1、p-NF-κBp65,上调IκBα水平(P<0.05)。结论:岩藻多糖抑制capan-1细胞增殖,对体内移植瘤抑瘤效果显著,其机制与调控G3BP1/NF-κB信号通路有关。展开更多
BACKGROUND The increased expression of G3BP1 was positively correlated with the prognosis of liver failure.AIM To investigate the effect of G3BP1 on the prognosis of acute liver failure(ALF)and acute-on-chronic liver ...BACKGROUND The increased expression of G3BP1 was positively correlated with the prognosis of liver failure.AIM To investigate the effect of G3BP1 on the prognosis of acute liver failure(ALF)and acute-on-chronic liver failure(ACLF)after the treatment of artificial liver support system(ALSS).METHODS A total of 244 patients with ALF and ACLF were enrolled in this study.The levels of G3BP1 on admission and at discharge were detected.The validation set of 514 patients was collected to verify the predicted effect of G3BP1 and the viability of prognosis.RESULTS This study was shown that lactate dehydrogenase(LDH),alpha-fetoprotein(AFP)and prothrombin time were closely related to the prognosis of patients.After the ALSS treatment,the patient’amount of decreased G3BP1 index in difference of G3BP1 between the value of discharge and admission(difG3BP1)<0 group had a nearly 10-fold increased risk of progression compared with the amount of increased G3BP1 index.The subgroup analysis showed that the difG3BP1<0 group had a higher risk of progression,regardless of model for end-stage liver disease high-risk or low-risk group.At the same time,compared with the inflam matory marks[tumor necrosis factor-α,interleukin(IL)-1βand IL-18],G3BP1 had higher discrimination and was more stable in the model analysis and validation set.When combined with AFP and LDH,concordance index was respectively 0.84 and 0.8 in training and validation cohorts.CONCLUSION This study indicated that G3BP1 could predict the prognosis of ALF or ACLF patients treated with ALSS.The combination of G3BP1,AFP and LDH could accurately evaluate the disease condition and predict the clinical endpoint of patients.展开更多
Stress granules(SGs)are membraneless cytoplasmic condensates that dynamically assemble in response to various stressors and reversibly disassemble after stimulus removal;however,the mechanisms underlying SG dynamics a...Stress granules(SGs)are membraneless cytoplasmic condensates that dynamically assemble in response to various stressors and reversibly disassemble after stimulus removal;however,the mechanisms underlying SG dynamics and their physiological roles in germ cell development are elusive.Here,we show that SERBP1(SERPINE1 mRNA binding protein 1)is a universal SG component and conserved regulator of SG clearance in somatic and male germ cells.SERBP1 interacts with the SG core component G3BP1 and 26S proteasome proteins PSMD10 and PSMA3 and recruits them to SGs.In the absence of SERBP1,reduced 20S proteasome activity,mislocalized valosin containing protein(VCP)and Fas associated factor family member 2(FAF2),and diminished K63-linked polyubiquitination of G3BP1 during the SG recovery period were observed.Interestingly,the depletion of SERBP1 in testicular cells in vivo causes increased germ cell apoptosis upon scrotal heat stress.Accordingly,we propose that a SERBP1-mediated mechanism regulates 26S proteasome activity and G3BP1 ubiquitination to facilitate SG clearance in both somatic and germ cell lines.展开更多
文摘目的研究G3BP1(Ras-GTPase-activating protein SH3 domain binding protein 1)在宫颈癌组织中的表达情况,分析G3BP1的表达水平及其相关临床意义。方法利用公开的GEO宫颈癌数据集分析G3BP1的mRNA水平在癌和癌旁组织的表达差异情况;取306例宫颈癌组织制作组织芯片,通过免疫组织化学检测癌组织中G3BP1的表达水平,统计分析G3BP1在宫颈癌中与正常宫颈组织和鳞状上皮内病变组织的表达差异情况,分析宫颈癌组织中G3BP1的表达与患者临床病理资料的相关性及对患者生存预后的影响。结果在GSE6791、GSE7803和GSE7410数据集中,我们发现G3BP1的mRNA水平在宫颈癌组织中的表达显著高于癌旁组织。免疫组织化学检测结果显示G3BP1的蛋白表达随着宫颈鳞状上皮内病变和宫颈癌的发生有逐渐增高的趋势。免疫组织化学和χ2检验结果显示G3BP1高表达与年龄、肿瘤大小、肿瘤分化、肌层浸润、阴道浸润、盆腔淋巴结转移、FIGO分期以及不良预后相关。Cox多因素分析显示G3BP1高表达是该批宫颈癌患者的独立危险预后因素。通过与G3BP1的表达水平整合可以显著促进FIGO分期在判断宫颈癌患者预后中的准确性。结论G3BP1高表达可能是宫颈癌患者的独立危险预后因素,并可能参与宫颈癌的发生发展等进程。
文摘目的:探讨岩藻多糖对胰腺癌的影响,并分析其机制。方法:MTT法分析细胞增殖抑制率,GEPIA数据库分析G3BP1在胰腺癌组织中的表达水平及与生存率的关系。qRT-PCR分析GTP酶激活蛋白结合蛋白1(Ras-GTPase-activating protein binding proteins,G3BP1)水平,Western blot法分析p-NF-κBp65、NF-κBp65和IκB-α水平。免疫共沉淀检测G3BP1与p-NF-κBp65之间相互作用。敲低或G3BP1过表达,观察其对岩藻多糖调控细胞增殖以及NF-κB信号通路的影响。裸鼠成瘤实验验证岩藻多糖对裸鼠体内瘤体的瘤重、瘤体积及G3BP1、p-NF-κBp65、NF-κBp65和IκBα水平的影响。结果:1~32μg/mL岩藻多糖抑制capan-1细胞增殖,岩藻多糖48 h IC_(50)为7.729μg/mL。G3BP1在胰腺癌肿瘤组织中的表达明显高于正常组织,G3BP1高表达患者的生存率低于G3BP1低表达患者。2、4、8μg/mL岩藻多糖能下调G3BP1、p-NF-κBp65,上调IκBα水平。Co-IP实验发现G3BP1与p-NF-κBp65相互结合,并且岩藻多糖作用后结合能力降低。敲低G3BP1能促进岩藻多糖抑制capan-1细胞增殖,下调G3BP1、p-NF-κBp65,上调IκBα水平(P<0.05);G3BP过表达能下调岩藻多糖抑制capan-1细胞增殖效果,上调G3BP1、p-NF-κBp65,下调IκBα水平(P<0.05)。体内实验显示,敲低G3BP1能促进岩藻多糖减少瘤体体积、瘤体质量,下调瘤体G3BP1、p-NF-κBp65,上调IκBα水平(P<0.05)。结论:岩藻多糖抑制capan-1细胞增殖,对体内移植瘤抑瘤效果显著,其机制与调控G3BP1/NF-κB信号通路有关。
文摘BACKGROUND The increased expression of G3BP1 was positively correlated with the prognosis of liver failure.AIM To investigate the effect of G3BP1 on the prognosis of acute liver failure(ALF)and acute-on-chronic liver failure(ACLF)after the treatment of artificial liver support system(ALSS).METHODS A total of 244 patients with ALF and ACLF were enrolled in this study.The levels of G3BP1 on admission and at discharge were detected.The validation set of 514 patients was collected to verify the predicted effect of G3BP1 and the viability of prognosis.RESULTS This study was shown that lactate dehydrogenase(LDH),alpha-fetoprotein(AFP)and prothrombin time were closely related to the prognosis of patients.After the ALSS treatment,the patient’amount of decreased G3BP1 index in difference of G3BP1 between the value of discharge and admission(difG3BP1)<0 group had a nearly 10-fold increased risk of progression compared with the amount of increased G3BP1 index.The subgroup analysis showed that the difG3BP1<0 group had a higher risk of progression,regardless of model for end-stage liver disease high-risk or low-risk group.At the same time,compared with the inflam matory marks[tumor necrosis factor-α,interleukin(IL)-1βand IL-18],G3BP1 had higher discrimination and was more stable in the model analysis and validation set.When combined with AFP and LDH,concordance index was respectively 0.84 and 0.8 in training and validation cohorts.CONCLUSION This study indicated that G3BP1 could predict the prognosis of ALF or ACLF patients treated with ALSS.The combination of G3BP1,AFP and LDH could accurately evaluate the disease condition and predict the clinical endpoint of patients.
基金This work,in part,was supported by grants from the National Natural Science Foundation of China(82171605 to S.Y.and 31900511 to F.W.)the Fundamental Research Funds for the Central Universities,Huazhong University of Science and Technology(No.2019kfyXJJS089 to F.W.)。
文摘Stress granules(SGs)are membraneless cytoplasmic condensates that dynamically assemble in response to various stressors and reversibly disassemble after stimulus removal;however,the mechanisms underlying SG dynamics and their physiological roles in germ cell development are elusive.Here,we show that SERBP1(SERPINE1 mRNA binding protein 1)is a universal SG component and conserved regulator of SG clearance in somatic and male germ cells.SERBP1 interacts with the SG core component G3BP1 and 26S proteasome proteins PSMD10 and PSMA3 and recruits them to SGs.In the absence of SERBP1,reduced 20S proteasome activity,mislocalized valosin containing protein(VCP)and Fas associated factor family member 2(FAF2),and diminished K63-linked polyubiquitination of G3BP1 during the SG recovery period were observed.Interestingly,the depletion of SERBP1 in testicular cells in vivo causes increased germ cell apoptosis upon scrotal heat stress.Accordingly,we propose that a SERBP1-mediated mechanism regulates 26S proteasome activity and G3BP1 ubiquitination to facilitate SG clearance in both somatic and germ cell lines.
