目的采用多种生物信息学分析工具和免疫组织化学染色,明确序列相似性20家族成员A(Family with sequence similarity 20 member A,FAM20A)在甲状腺癌(Thyroid carcinoma,TC)中的表达及临床诊断意义。方法通过癌症基因组图谱(TCGA)数据库...目的采用多种生物信息学分析工具和免疫组织化学染色,明确序列相似性20家族成员A(Family with sequence similarity 20 member A,FAM20A)在甲状腺癌(Thyroid carcinoma,TC)中的表达及临床诊断意义。方法通过癌症基因组图谱(TCGA)数据库获取甲状腺癌相关数据,分析FAM20A在甲状腺癌组织和正常组织之间的表达差异,绘制ROC曲线计算AUC值,评估FAM20A对甲状腺癌的诊断效能。利用GEO数据库下载GSE205733甲状腺癌数据集,分析FAM20A基因在甲状腺癌组织中的表达水平,并进行基因富集分析(Gene set enrichment analysis,GSEA);利用在线数据库STRING分析FAM20A的蛋白相互作用网络。同时收集2022年3月—2023年2月期间于首都医科大学附属北京潞河医院甲状腺外科行甲状腺结节手术的29例患者病理组织样本,进行病理组织石蜡切片和免疫组织化学染色,验证比较甲状腺癌及良性甲状腺结节样本中FAM20A的表达水平。结果TCGA数据库结果表明,与正常甲状腺组织相比,FAM20A在甲状腺癌组织中的表达显著升高(P<0.001)。FAM20A表达水平与甲状腺癌临床TNM分期和病理分型相关(P<0.05)。FAM20A对于甲状腺癌的诊断效能显著(AUC=0.828)。GSE205733数据分析显示与正常组织相比,FAM20A在甲状腺癌组织中表达显著增加(P<0.001)。GSEA富集分析结果显示FAM20A主要与P53和白介素等信号传导通路相关。STRING数据库分析显示与FAM20A相互作用的蛋白主要有PEAK,FAM20C和ENAM等。免疫组织化学染色结果显示FAM20A在甲状腺癌组织中表达显著增加(P<0.05)。结论FAM20A基因在甲状腺癌组织中高表达,与患者临床TNM分期和病理分型相关,可作为诊断甲状腺癌的潜在分子标记物,且与P53和白介素等信号传导通路相关,为研究甲状腺癌进展分子机制及靶向药物的开发提供理论依据。展开更多
Enamelin （ENAM） has three putative phosphoserines （pSers） phosphorylated by a Golgi-associated secretory pathway kinase （FAM20C） based on their distinctive Ser-x-Glu （S-x-E） motifs. Fam2OC-knockout mice show s...Enamelin （ENAM） has three putative phosphoserines （pSers） phosphorylated by a Golgi-associated secretory pathway kinase （FAM20C） based on their distinctive Ser-x-Glu （S-x-E） motifs. Fam2OC-knockout mice show severe enamel defects similar to those in the Enam-knockout mice, implying an important role of the pSers in ENAM. To determine the role of pSer5s in ENAM, we characterized ENAMRgsc514 mice, in which Sers5 cannot be phosphorylated by FAM20C due to an E57〉Gs7 mutation in the S-x-E motif, The enamel microstructure of 4-week-old mice was examined by scanning electron microscopy. The teeth of 6-day-old mice were characterized by histology and immunohistochemistry. The protein lysates of the first lower molars of 4-day-old mice were analyzed by Western immunoblotting using antibodies against ENAM, ameloblastin and amelogenin. ENAMRgsc514 heterozygotes showed a disorganized enamel microstructure, while the homozygotes had no enamel on the dentin surface. The N-terminal fragments of ENAM in the heterozygotes were detained in the ameloblasts and localized in the mineralization front of enamel matrix, while those in the WT mice were secreted out of ameloblasts and distributed evenly in the outer 1/2 of enamel matrix. Surprisingly, the 15 kDa C-terminal fragments of ameloblastin were not detected in the molar lysates of the homozygotes. These results suggest that the phosphorylation of SerSS may be an essential posttranslational modification of ENAM and is required for the interaction with other enamel matrix molecules such as ameloblastin in mediating the structural organization of enamel matrix and protein-mineral interactions during enamel formation.展开更多
目的观察小鼠髁突发育过程的不同阶段,FAM20C在髁突软骨中的时空表达特点,试探究其在小鼠髁突发育过程中的可能作用机制。方法苏木精-伊红(HE)染色和改良番红O-固绿染色观察胚胎17.5 d和出生后0、7、21 d 4组小鼠髁突软骨及软骨下骨的...目的观察小鼠髁突发育过程的不同阶段,FAM20C在髁突软骨中的时空表达特点,试探究其在小鼠髁突发育过程中的可能作用机制。方法苏木精-伊红(HE)染色和改良番红O-固绿染色观察胚胎17.5 d和出生后0、7、21 d 4组小鼠髁突软骨及软骨下骨的形态学变化。免疫组化染色观察相应时间点FAM20C在小鼠髁突软骨组织中的定位及表达。测量FAM20C阳性表达的平均光密度值,并进行单因素方差分析。结果HE染色和改良番红O-固绿染色结果表明:随着软骨内骨化的进展,髁突软骨细胞层长度减少,软骨下骨体积增加,下颌髁突体积增大;免疫组化结果表明:4组小鼠髁突软骨组织中均有FAM20C阳性表达,FAM20C主要表达于髁突软骨增殖软骨细胞层和前肥大软骨细胞层,少量表达于肥大软骨细胞层及软骨下骨层,随着髁突发育,FAM20C表达逐渐减少。统计学结果显示,各时间点FAM20C阳性表达差异有统计学意义(P<0.01)。结论FAM20C参与小鼠下颌髁突发育,可能通过调节髁突软骨细胞的增殖和分化在髁突形成中发挥重要作用。展开更多
Gliomas are the most prevalent and aggressive malignancies of the nervous system.Previous bioinformatic studies have revealed the crucial role of the secretory pathway kinase FAM20C in the prediction of glioma invasio...Gliomas are the most prevalent and aggressive malignancies of the nervous system.Previous bioinformatic studies have revealed the crucial role of the secretory pathway kinase FAM20C in the prediction of glioma invasion and malignancy.However,little is known about the pathogenesis of FAM20C in the regulation of glioma.Here,we construct the full-length transcriptome atlas in paired gliomas and observe that 22 genes are upregulated by full-length transcriptome and differential APA analysis.Analysis of ATAC-seq data reveals that both FAM20C and NPTN are the hub genes with chromatin openness and differential expression.Further,in vitro and in vivo studies suggest that FAM20C stimulates the proliferation and metastasis of glioma cells.Meanwhile,NPTN,a novel cancer suppressor gene,counteracts the function of FAM20C by inhibiting both the proliferation and migration of glioma.The blockade of FAM20C by neutralizing antibodies results in the regression of xenograft tumors.Moreover,MAX,BRD4,MYC,and REST are found to be the potential trans-active factors for the regulation of FAM20C.Taken together,our results uncover the oncogenic role of FAM20C in glioma and shed new light on the treatment of glioma by abolishing FAM20C.展开更多
Family with sequence similarity 20, member C （FAM20C） 又称牙本质基质蛋白4（DMP4），是一种分泌性钙结合激酶，在矿化组织中高表达，可使生物矿化相关的分泌性钙结合磷酸蛋白（scPPs）家族磷酸化，调控磷酸钙沉积，形成并影响羟...Family with sequence similarity 20, member C （FAM20C） 又称牙本质基质蛋白4（DMP4），是一种分泌性钙结合激酶，在矿化组织中高表达，可使生物矿化相关的分泌性钙结合磷酸蛋白（scPPs）家族磷酸化，调控磷酸钙沉积，形成并影响羟基磷灰石晶体的生长和排列。FAM20C突变可致Raine综合征（又称硬化性骨发育不良），导致骨、牙发育缺陷，低磷酸血症等，新生儿致死率高。近年来研究表明，FAM20C可促进成骨细胞、成釉细胞及成牙本质细胞分化，负性调控成纤维细胞生长因子23（FGF23）影响血磷稳态，在骨和牙齿发育矿化中发挥重要作用。就FAM20C在骨和牙发育矿化中的研究现状作一综述。展开更多
文摘目的采用多种生物信息学分析工具和免疫组织化学染色,明确序列相似性20家族成员A(Family with sequence similarity 20 member A,FAM20A)在甲状腺癌(Thyroid carcinoma,TC)中的表达及临床诊断意义。方法通过癌症基因组图谱(TCGA)数据库获取甲状腺癌相关数据,分析FAM20A在甲状腺癌组织和正常组织之间的表达差异,绘制ROC曲线计算AUC值,评估FAM20A对甲状腺癌的诊断效能。利用GEO数据库下载GSE205733甲状腺癌数据集,分析FAM20A基因在甲状腺癌组织中的表达水平,并进行基因富集分析(Gene set enrichment analysis,GSEA);利用在线数据库STRING分析FAM20A的蛋白相互作用网络。同时收集2022年3月—2023年2月期间于首都医科大学附属北京潞河医院甲状腺外科行甲状腺结节手术的29例患者病理组织样本,进行病理组织石蜡切片和免疫组织化学染色,验证比较甲状腺癌及良性甲状腺结节样本中FAM20A的表达水平。结果TCGA数据库结果表明,与正常甲状腺组织相比,FAM20A在甲状腺癌组织中的表达显著升高(P<0.001)。FAM20A表达水平与甲状腺癌临床TNM分期和病理分型相关(P<0.05)。FAM20A对于甲状腺癌的诊断效能显著(AUC=0.828)。GSE205733数据分析显示与正常组织相比,FAM20A在甲状腺癌组织中表达显著增加(P<0.001)。GSEA富集分析结果显示FAM20A主要与P53和白介素等信号传导通路相关。STRING数据库分析显示与FAM20A相互作用的蛋白主要有PEAK,FAM20C和ENAM等。免疫组织化学染色结果显示FAM20A在甲状腺癌组织中表达显著增加(P<0.05)。结论FAM20A基因在甲状腺癌组织中高表达,与患者临床TNM分期和病理分型相关,可作为诊断甲状腺癌的潜在分子标记物,且与P53和白介素等信号传导通路相关,为研究甲状腺癌进展分子机制及靶向药物的开发提供理论依据。
基金supported by NIH grant DE026461start funding of Texas A&M University College of Dentistry
文摘Enamelin （ENAM） has three putative phosphoserines （pSers） phosphorylated by a Golgi-associated secretory pathway kinase （FAM20C） based on their distinctive Ser-x-Glu （S-x-E） motifs. Fam2OC-knockout mice show severe enamel defects similar to those in the Enam-knockout mice, implying an important role of the pSers in ENAM. To determine the role of pSer5s in ENAM, we characterized ENAMRgsc514 mice, in which Sers5 cannot be phosphorylated by FAM20C due to an E57〉Gs7 mutation in the S-x-E motif, The enamel microstructure of 4-week-old mice was examined by scanning electron microscopy. The teeth of 6-day-old mice were characterized by histology and immunohistochemistry. The protein lysates of the first lower molars of 4-day-old mice were analyzed by Western immunoblotting using antibodies against ENAM, ameloblastin and amelogenin. ENAMRgsc514 heterozygotes showed a disorganized enamel microstructure, while the homozygotes had no enamel on the dentin surface. The N-terminal fragments of ENAM in the heterozygotes were detained in the ameloblasts and localized in the mineralization front of enamel matrix, while those in the WT mice were secreted out of ameloblasts and distributed evenly in the outer 1/2 of enamel matrix. Surprisingly, the 15 kDa C-terminal fragments of ameloblastin were not detected in the molar lysates of the homozygotes. These results suggest that the phosphorylation of SerSS may be an essential posttranslational modification of ENAM and is required for the interaction with other enamel matrix molecules such as ameloblastin in mediating the structural organization of enamel matrix and protein-mineral interactions during enamel formation.
文摘目的观察小鼠髁突发育过程的不同阶段,FAM20C在髁突软骨中的时空表达特点,试探究其在小鼠髁突发育过程中的可能作用机制。方法苏木精-伊红(HE)染色和改良番红O-固绿染色观察胚胎17.5 d和出生后0、7、21 d 4组小鼠髁突软骨及软骨下骨的形态学变化。免疫组化染色观察相应时间点FAM20C在小鼠髁突软骨组织中的定位及表达。测量FAM20C阳性表达的平均光密度值,并进行单因素方差分析。结果HE染色和改良番红O-固绿染色结果表明:随着软骨内骨化的进展,髁突软骨细胞层长度减少,软骨下骨体积增加,下颌髁突体积增大;免疫组化结果表明:4组小鼠髁突软骨组织中均有FAM20C阳性表达,FAM20C主要表达于髁突软骨增殖软骨细胞层和前肥大软骨细胞层,少量表达于肥大软骨细胞层及软骨下骨层,随着髁突发育,FAM20C表达逐渐减少。统计学结果显示,各时间点FAM20C阳性表达差异有统计学意义(P<0.01)。结论FAM20C参与小鼠下颌髁突发育,可能通过调节髁突软骨细胞的增殖和分化在髁突形成中发挥重要作用。
基金supported by the grants from Department of Science and Technology of Sichuan Province,China(No.23ZDYF2212)Medico-Engineering Cooperation Funds from the University of Electronic Science and Technology of China(No.ZYGX2021YGLH209)+1 种基金the Department of Science and Technology of Sichuan Province(Nos.2022NSFSC0760 and 2022JDTD0024)the Chengdu Science and Technology Bureau(2022-YF05-01625-SN)。
文摘Gliomas are the most prevalent and aggressive malignancies of the nervous system.Previous bioinformatic studies have revealed the crucial role of the secretory pathway kinase FAM20C in the prediction of glioma invasion and malignancy.However,little is known about the pathogenesis of FAM20C in the regulation of glioma.Here,we construct the full-length transcriptome atlas in paired gliomas and observe that 22 genes are upregulated by full-length transcriptome and differential APA analysis.Analysis of ATAC-seq data reveals that both FAM20C and NPTN are the hub genes with chromatin openness and differential expression.Further,in vitro and in vivo studies suggest that FAM20C stimulates the proliferation and metastasis of glioma cells.Meanwhile,NPTN,a novel cancer suppressor gene,counteracts the function of FAM20C by inhibiting both the proliferation and migration of glioma.The blockade of FAM20C by neutralizing antibodies results in the regression of xenograft tumors.Moreover,MAX,BRD4,MYC,and REST are found to be the potential trans-active factors for the regulation of FAM20C.Taken together,our results uncover the oncogenic role of FAM20C in glioma and shed new light on the treatment of glioma by abolishing FAM20C.
文摘Family with sequence similarity 20, member C （FAM20C） 又称牙本质基质蛋白4（DMP4），是一种分泌性钙结合激酶，在矿化组织中高表达，可使生物矿化相关的分泌性钙结合磷酸蛋白（scPPs）家族磷酸化，调控磷酸钙沉积，形成并影响羟基磷灰石晶体的生长和排列。FAM20C突变可致Raine综合征（又称硬化性骨发育不良），导致骨、牙发育缺陷，低磷酸血症等，新生儿致死率高。近年来研究表明，FAM20C可促进成骨细胞、成釉细胞及成牙本质细胞分化，负性调控成纤维细胞生长因子23（FGF23）影响血磷稳态，在骨和牙齿发育矿化中发挥重要作用。就FAM20C在骨和牙发育矿化中的研究现状作一综述。
