Purpose:Approximately 1/3 of brain tumors are gliomas.Previous glioma-related studies have reported increased expression of periostin(POSTN)in these cancerous tissues,but the role and mechanism of POSTN in glioma deve...Purpose:Approximately 1/3 of brain tumors are gliomas.Previous glioma-related studies have reported increased expression of periostin(POSTN)in these cancerous tissues,but the role and mechanism of POSTN in glioma development remain unclear.Methods:Nanoscale liquid chromatography coupled with tandem mass spectrometry(nano LC-MS/MS)and RNA sequencing were used to identify diferential protein and mRNA expression in clinical glioma samples.Quantitative real-time PCR(qRT-PCR)was used to measure the expression of POSTN in tissues and cells.The efects of POSTN on glioma cell migration and invasion were examined using wound healing,Transwell,and three-dimensional spheroid assays in vitro and a nude mouse xenograft model in vivo.The efects of POSTN on the stability,endocytosis,and degradation of EGFR were examined by immunoblotting and immunofuorescence staining.Truncation mutation analysis was performed to investigate direct interactions between POSTN and EGFR.Immunohistochemical staining was carried out to confrm the clinical signifcance of POSTN.Results:Overexpression of POSTN induced epithelial-to-mesenchymal transition(EMT)in glioma cells in vivo and in vitro.Mechanistically,POSTN downregulation inhibited EGFR signaling by promoting EGFR endocytosis and degradation.In addition,POSTN was found to bind to EGFR and RIN1,inhibiting EGFR endocytosis and degradation and thus activating the PI3K-Akt signaling pathway.Conclusion:These fndings indicate the mechanism by which the POSTN/EGFR/RIN1 axis inhibits EGFR endocytosis and degradation,resulting in glioma cell EMT through the PI3K-AKT signaling pathway.Targeting POSTN/EGFR/RIN1 interactions may guarantee benefcial outcomes of glioma treatment.展开更多
目的观察胃癌相关蛋白质POSTN在胃黏膜癌变过程中的表达与意义,为胃癌的早期诊断、病理分型与评估预后提供有价值的实验依据。方法收集正常胃黏膜、癌旁及胃癌组织标本8例,采用Western blotting检测POSTN蛋白在上述组织中的表达;其次利...目的观察胃癌相关蛋白质POSTN在胃黏膜癌变过程中的表达与意义,为胃癌的早期诊断、病理分型与评估预后提供有价值的实验依据。方法收集正常胃黏膜、癌旁及胃癌组织标本8例,采用Western blotting检测POSTN蛋白在上述组织中的表达;其次利用免疫组化染色观察POSTN蛋白在正常胃黏膜、癌旁及胃癌组织中的表达,并分析其在胃黏膜癌变过程中表达的意义。结果 Western blotting结果显示,胃癌组织中POSTN蛋白表达明显高于正常胃黏膜和癌旁组织(P<0.01),同时癌旁组织中的表达高于正常胃黏膜组织(P<0.01)。免疫组化结果显示,正常胃黏膜、癌旁和胃癌组织中POSTN蛋白表达的阳性率分别为14.71%、46.15%和77.48%,而胃高、中和低分化腺癌中表达的阳性率分别为60%、70.6%和81.4%。POSTN蛋白表达与胃癌组织的分化程度相关,即组织分化降低,其表达增强(P<0.01)。结论胃癌组织中POSTN蛋白表达高于正常胃黏膜和癌旁组织,癌旁组织中的表达高于正常胃黏膜组织,该蛋白表达与胃癌组织的发生和分化程度有关。展开更多
目的通过对1例智力发育迟缓患儿全外显子组测序(WES),旨在为该患儿寻找潜在的致病原因。方法纳入1例在复旦大学附属儿科医院(我院)住院期间诊断不明患儿,采用Sure Selct Human All Exon捕获试剂盒和Illumina Hi Seq2000测序平台,行WES检...目的通过对1例智力发育迟缓患儿全外显子组测序(WES),旨在为该患儿寻找潜在的致病原因。方法纳入1例在复旦大学附属儿科医院(我院)住院期间诊断不明患儿,采用Sure Selct Human All Exon捕获试剂盒和Illumina Hi Seq2000测序平台,行WES检测;数据分析采用我院分子诊断中心建立的高通量测序数据分析流程;结果采用Sanger直接测序法进行验证。建立体外大鼠神经元原代培养和糖氧剥夺(OGD)模型,分为空白对照组(Sham+PBS亚组、Sham+POSTN亚组)、OGD组(OGD+PBS亚组和OGD+POSTN亚组),采用免疫荧光、LDH毒性检测、Brd U和TUNEL方法检测POSTN对2组及其亚组神经元生长增殖和凋亡的影响。结果患儿WES共检测到532 151个变异,筛选后检测到POSTN基因(NM_006475)外显子23:c.A2475T:p.G825G和外显子19:c.G2251A:p.E751K。Sanger测序显示p.G825G来自母亲,p.E751K来自父亲,符合复合杂合遗传模式。培养的大鼠原代神经元(MAP-2或Neu N阳性细胞)中有内源性POSTN的表达。LDH活性Sham+POSTN亚组与Sham+PBS亚组差异无统计学意义,OGD+POSTN亚组与OGD+PBS亚组差异有统计学意义。OGD+PBS亚组致神经元细胞缺氧2、6、12 d后比Sham+PBS亚组Brd U阳性的神经元数量增高,TUNEL阳性神经元凋亡也增高;OGD+POSTN亚组各时间点神经元增殖较OGD+PBS亚组均明显增高,但凋亡减少。结论 POSTN可以减少脑损伤后LDH对于神经元的细胞毒性,促进损伤后神经元的增殖,抑制凋亡。展开更多
基金supported by grants from National Natural Science Foundation of China (81772682)the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).
文摘Purpose:Approximately 1/3 of brain tumors are gliomas.Previous glioma-related studies have reported increased expression of periostin(POSTN)in these cancerous tissues,but the role and mechanism of POSTN in glioma development remain unclear.Methods:Nanoscale liquid chromatography coupled with tandem mass spectrometry(nano LC-MS/MS)and RNA sequencing were used to identify diferential protein and mRNA expression in clinical glioma samples.Quantitative real-time PCR(qRT-PCR)was used to measure the expression of POSTN in tissues and cells.The efects of POSTN on glioma cell migration and invasion were examined using wound healing,Transwell,and three-dimensional spheroid assays in vitro and a nude mouse xenograft model in vivo.The efects of POSTN on the stability,endocytosis,and degradation of EGFR were examined by immunoblotting and immunofuorescence staining.Truncation mutation analysis was performed to investigate direct interactions between POSTN and EGFR.Immunohistochemical staining was carried out to confrm the clinical signifcance of POSTN.Results:Overexpression of POSTN induced epithelial-to-mesenchymal transition(EMT)in glioma cells in vivo and in vitro.Mechanistically,POSTN downregulation inhibited EGFR signaling by promoting EGFR endocytosis and degradation.In addition,POSTN was found to bind to EGFR and RIN1,inhibiting EGFR endocytosis and degradation and thus activating the PI3K-Akt signaling pathway.Conclusion:These fndings indicate the mechanism by which the POSTN/EGFR/RIN1 axis inhibits EGFR endocytosis and degradation,resulting in glioma cell EMT through the PI3K-AKT signaling pathway.Targeting POSTN/EGFR/RIN1 interactions may guarantee benefcial outcomes of glioma treatment.
文摘目的观察胃癌相关蛋白质POSTN在胃黏膜癌变过程中的表达与意义,为胃癌的早期诊断、病理分型与评估预后提供有价值的实验依据。方法收集正常胃黏膜、癌旁及胃癌组织标本8例,采用Western blotting检测POSTN蛋白在上述组织中的表达;其次利用免疫组化染色观察POSTN蛋白在正常胃黏膜、癌旁及胃癌组织中的表达,并分析其在胃黏膜癌变过程中表达的意义。结果 Western blotting结果显示,胃癌组织中POSTN蛋白表达明显高于正常胃黏膜和癌旁组织(P<0.01),同时癌旁组织中的表达高于正常胃黏膜组织(P<0.01)。免疫组化结果显示,正常胃黏膜、癌旁和胃癌组织中POSTN蛋白表达的阳性率分别为14.71%、46.15%和77.48%,而胃高、中和低分化腺癌中表达的阳性率分别为60%、70.6%和81.4%。POSTN蛋白表达与胃癌组织的分化程度相关,即组织分化降低,其表达增强(P<0.01)。结论胃癌组织中POSTN蛋白表达高于正常胃黏膜和癌旁组织,癌旁组织中的表达高于正常胃黏膜组织,该蛋白表达与胃癌组织的发生和分化程度有关。
文摘目的通过对1例智力发育迟缓患儿全外显子组测序(WES),旨在为该患儿寻找潜在的致病原因。方法纳入1例在复旦大学附属儿科医院(我院)住院期间诊断不明患儿,采用Sure Selct Human All Exon捕获试剂盒和Illumina Hi Seq2000测序平台,行WES检测;数据分析采用我院分子诊断中心建立的高通量测序数据分析流程;结果采用Sanger直接测序法进行验证。建立体外大鼠神经元原代培养和糖氧剥夺(OGD)模型,分为空白对照组(Sham+PBS亚组、Sham+POSTN亚组)、OGD组(OGD+PBS亚组和OGD+POSTN亚组),采用免疫荧光、LDH毒性检测、Brd U和TUNEL方法检测POSTN对2组及其亚组神经元生长增殖和凋亡的影响。结果患儿WES共检测到532 151个变异,筛选后检测到POSTN基因(NM_006475)外显子23:c.A2475T:p.G825G和外显子19:c.G2251A:p.E751K。Sanger测序显示p.G825G来自母亲,p.E751K来自父亲,符合复合杂合遗传模式。培养的大鼠原代神经元(MAP-2或Neu N阳性细胞)中有内源性POSTN的表达。LDH活性Sham+POSTN亚组与Sham+PBS亚组差异无统计学意义,OGD+POSTN亚组与OGD+PBS亚组差异有统计学意义。OGD+PBS亚组致神经元细胞缺氧2、6、12 d后比Sham+PBS亚组Brd U阳性的神经元数量增高,TUNEL阳性神经元凋亡也增高;OGD+POSTN亚组各时间点神经元增殖较OGD+PBS亚组均明显增高,但凋亡减少。结论 POSTN可以减少脑损伤后LDH对于神经元的细胞毒性,促进损伤后神经元的增殖,抑制凋亡。
