目的·探索RBX1(ring-box protein 1)在葡萄膜黑色素瘤(uveal melanoma,UVM)肿瘤细胞中对免疫相关基因的调控作用。方法·通过检索癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库分析RBX1在肿瘤中的表达水平以及与临床分...目的·探索RBX1(ring-box protein 1)在葡萄膜黑色素瘤(uveal melanoma,UVM)肿瘤细胞中对免疫相关基因的调控作用。方法·通过检索癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库分析RBX1在肿瘤中的表达水平以及与临床分期、生存预后的相关性。使用靶向RBX1的小干扰RNA(small interfering RNA,siRNA)分别在UVM细胞系92.1、OMM2.3和MEL290中瞬时敲低RBX1,并对瞬时敲低RBX1的92.1细胞进行转录组测序,分析si RBX1转染细胞与对照细胞的差异表达基因,并采用基因集富集分析(gene set enrichment analysis,GSEA)对差异基因进行分析,探究RBX1与肿瘤免疫相关基因的关系。在分析结果的基础上,通过实时荧光定量PCR(qPCR)分别检测瞬时敲低RBX1的92.1、OMM2.3和MEL290细胞系中信号转导与转录激活因子1(signal transducer and activator of transcription 1,STAT1)及其下游的CXC趋化因子配体9(C-X-C motif chemokine ligand 9,CXCL9)和CXCL10的mRNA表达水平,并通过Western blotting检测92.1细胞中STAT1及p-STAT1蛋白表达水平。在瞬时敲低RBX1的细胞系OMM2.3和MEL290中,分别加入5 nmol/L或10 nmol/L的STAT1抑制剂fludarabine,处理48 h后通过qPCR检测CXCL9和CXCL10的mRNA表达水平。结果·TCGA数据库分析表明:与正常组织相比,RBX1在多种肿瘤中高表达,在肾上腺皮质癌和UVM中显著高表达;且这2种肿瘤分期晚的患者,RBX1表达水平更高,同时RBX1表达水平高的患者的总生存期更短(均P<0.05)。对瞬时敲低RBX1的92.1细胞和对照细胞进行转录组测序,获得差异基因,并且GSEA结果显示,RBX1参与调控肿瘤免疫相关通路。热图分析显示RBX1敲低后STAT1表达水平上升。在92.1、OMM2.3和MEL290细胞系中,qPCR结果显示RBX1敲低后STAT1、CXCL9和CXCL10的mRNA表达水平上升,Western blotting结果显示在92.1细胞系中敲低RBX1后STAT1及p-STAT1表达水平上升。加入STAT1抑制剂后,OMM2.3和MEL290细胞系中的CXCL9和CXCL10 mRNA上调表达被抑制。结论·RBX1在UVM细胞中可能通过STAT1调控CXCL9和CXCL10的表达,参与肿瘤免疫调控。展开更多
SAG(Sensitive to Apoptosis Gene),also known as RBX2(RING box protein 2),ROC2(Regulator of Cullins 2),or RNF7(RING Finger Protein 7),was originally cloned in our laboratory as a redox inducible antioxi-dant protein and...SAG(Sensitive to Apoptosis Gene),also known as RBX2(RING box protein 2),ROC2(Regulator of Cullins 2),or RNF7(RING Finger Protein 7),was originally cloned in our laboratory as a redox inducible antioxi-dant protein and later characterized as the second member of the RBX/ROC RING component of the SCF(SKP1-CUL-F-box Proteins)E3 ubiquitin ligase.When acting alone,SAG scavenges oxygen radicals by forming inter-and intra-molecular disulfide bonds,whereas by forming a complex with other components of the SCF E3 ligase,SAG promotes ubiquitination and degradation of a number of protein substrates,includ-ing c-JUN,DEPTOR,HIF-1α,IκBα,NF1,NOXA,p27,and procaspase-3,thus regulating various signaling path-ways and biological processes.Specifically,SAG pro-tects cells from apoptosis,confers radioresistance,and plays an essential and non-redundant role in mouse embryogenesis and vasculogenesis.Furthermore,stress-inducible SAG is overexpressed in a number of human cancers and SAG overexpression correlates with poor patient prognosis.Finally,SAG transgenic expression in epidermis causes an early stage inhibi-tion,but later stage promotion,of skin tumorigenesis triggered by DMBA/TPA.Given its major role in pro-moting targeted degradation of tumor suppressive proteins,leading to apoptosis suppression and accel-erated tumorigenesis,SAG E3 ligase appears to be an attractive anticancer target.展开更多
目的探讨前牙拔牙位点即刻保存中重组合异种骨(reconstituted bone xenograft,RBX)与羟基磷灰石生物陶瓷复合物的作用。方法比格犬共6只,随机分为A、B、C组,每组各2只。所有动物拔除上颌全部第二切牙后,三组采用不同的实验方法,A组应用...目的探讨前牙拔牙位点即刻保存中重组合异种骨(reconstituted bone xenograft,RBX)与羟基磷灰石生物陶瓷复合物的作用。方法比格犬共6只,随机分为A、B、C组,每组各2只。所有动物拔除上颌全部第二切牙后,三组采用不同的实验方法,A组应用RBX与羟基磷灰石生物陶瓷复合物,B组仅应用羟基磷灰石生物陶瓷,C组为对照组,术后4周每组各处死1只犬,12周每组再处死1只犬,观察大体标本、形态学测量以及CT检查评价拔牙窝愈合情况。结果三组犬4、12周时唇舌侧牙槽嵴宽度差值比较差异均有统计学意义(P<0.05),A组差值最小;4周与12周时拔牙窝唇侧骨吸收程度A组阻止牙槽骨吸收效果最好,B组次之,C组最差;12周时A组唇侧牙槽嵴高度差值比较优于B组和C组(P<0.05);术后4周与12周CT值与术前CT值之差A组均高于B组和C组(P<0.05)。结论 RBX与羟基磷灰石生物陶瓷复合物联合应用,可减缓牙槽嵴吸收,促进新生骨形成,具有保存拔牙位点的作用,可作为良好的新型复合骨移植材料。展开更多
基金supported by the NCI grants(CA118762 and CA156744)to Yi Sun.
文摘SAG(Sensitive to Apoptosis Gene),also known as RBX2(RING box protein 2),ROC2(Regulator of Cullins 2),or RNF7(RING Finger Protein 7),was originally cloned in our laboratory as a redox inducible antioxi-dant protein and later characterized as the second member of the RBX/ROC RING component of the SCF(SKP1-CUL-F-box Proteins)E3 ubiquitin ligase.When acting alone,SAG scavenges oxygen radicals by forming inter-and intra-molecular disulfide bonds,whereas by forming a complex with other components of the SCF E3 ligase,SAG promotes ubiquitination and degradation of a number of protein substrates,includ-ing c-JUN,DEPTOR,HIF-1α,IκBα,NF1,NOXA,p27,and procaspase-3,thus regulating various signaling path-ways and biological processes.Specifically,SAG pro-tects cells from apoptosis,confers radioresistance,and plays an essential and non-redundant role in mouse embryogenesis and vasculogenesis.Furthermore,stress-inducible SAG is overexpressed in a number of human cancers and SAG overexpression correlates with poor patient prognosis.Finally,SAG transgenic expression in epidermis causes an early stage inhibi-tion,but later stage promotion,of skin tumorigenesis triggered by DMBA/TPA.Given its major role in pro-moting targeted degradation of tumor suppressive proteins,leading to apoptosis suppression and accel-erated tumorigenesis,SAG E3 ligase appears to be an attractive anticancer target.
文摘目的探讨前牙拔牙位点即刻保存中重组合异种骨(reconstituted bone xenograft,RBX)与羟基磷灰石生物陶瓷复合物的作用。方法比格犬共6只,随机分为A、B、C组,每组各2只。所有动物拔除上颌全部第二切牙后,三组采用不同的实验方法,A组应用RBX与羟基磷灰石生物陶瓷复合物,B组仅应用羟基磷灰石生物陶瓷,C组为对照组,术后4周每组各处死1只犬,12周每组再处死1只犬,观察大体标本、形态学测量以及CT检查评价拔牙窝愈合情况。结果三组犬4、12周时唇舌侧牙槽嵴宽度差值比较差异均有统计学意义(P<0.05),A组差值最小;4周与12周时拔牙窝唇侧骨吸收程度A组阻止牙槽骨吸收效果最好,B组次之,C组最差;12周时A组唇侧牙槽嵴高度差值比较优于B组和C组(P<0.05);术后4周与12周CT值与术前CT值之差A组均高于B组和C组(P<0.05)。结论 RBX与羟基磷灰石生物陶瓷复合物联合应用,可减缓牙槽嵴吸收,促进新生骨形成,具有保存拔牙位点的作用,可作为良好的新型复合骨移植材料。
