Purpose: To investigate the ferritin distribution in epithelial ovarian cancer patients according to the FIGO stage in the prognosis of epithelial ovarian cancer. Method: All ovarian cancer patients were divided into ...Purpose: To investigate the ferritin distribution in epithelial ovarian cancer patients according to the FIGO stage in the prognosis of epithelial ovarian cancer. Method: All ovarian cancer patients were divided into two groups according their FIGO stage. Benign ovarian tumor patients were analyzed as the control. Serum ferritin, serum iron, and other related medical index were detected by automatic instruments for all patients. In addition, ferritin heavy chain (FHC) and ferritin light chain (FLC) proteins were detected by immunohistochemical staining in 60 epithelial ovarian cancer (EOC) patients and 30 benign ovarian tumor (BOT) patients, which were diagnosed in our department between 2011 and 2016. Results: The serum ferritin concentration was significantly higher in the EOC group than in the BOT group (172.56 ± 99.39 ng/mL vs 78.18 ± 43.06 ng/mL;p μmol/L vs 14.92 ± 6.36 μmol/L;p p p p p p p > 0.05). Conclusion: Patients showed an overexpression of ferritin and a downregulation of serum iron correlated with the prognosis of epithelial ovarian cancer, which may be a new target for diagnosis and treatment of epithelial ovarian cancer.展开更多
目的研究四川白鹅铁蛋白重链(Ferritin heavy chain,FHC)基因编码区序列特征及其组织表达差异。方法应用分子生物学技术克隆FHC编码区序列并分析预测其编码蛋白质的结构和功能,实时荧光定量PCR技术定量检测四川白鹅13种组织中FHC的表达...目的研究四川白鹅铁蛋白重链(Ferritin heavy chain,FHC)基因编码区序列特征及其组织表达差异。方法应用分子生物学技术克隆FHC编码区序列并分析预测其编码蛋白质的结构和功能,实时荧光定量PCR技术定量检测四川白鹅13种组织中FHC的表达量。结果四川白鹅FHC编码区序列为549 bp,编码182个氨基酸,其蛋白质分子质量为21345.9,理论pI值为5.74。四川白鹅FHC氨基酸序列与北京鸭和原鸡的相似性最高(分别为100%和97%),与北京鸭具有最近的亲缘关系。FHC是稳定的可溶性蛋白质,含有高度保守的2个铁蛋白铁离子结合结构域标签和7个亚铁氧化酶双铁离子结合位点。在所检测的四川白鹅13种组织中,FHC基因均有表达,并且不同组织中FHC表达量存在明显差异。四川白鹅脾脏和视网膜中FHC的表达量显著高于其他各组织(P<0.05),肝脏、垂体和肾脏次之,肌肉中FHC表达量最低。结论四川白鹅FHC是高度保守的、组织特异性表达的基因,具有维持铁稳态和参与抗氧化的双重功能,这为阐明鹅FHC功能和铁代谢机制的研究奠定了理论基础。展开更多
[目的]探究ENO1通过NCOA4对胶质母细胞瘤(GBM)细胞铁死亡的调节机制。[方法]qRT-PCR和Western blot检测GBM组织和正常组织中ENO1的表达。人GBM细胞系U251细胞分为对照组、si-ENO1-NC组、si-ENO1-1组、si-ENO1-2组、si-ENO1+si-NCOA4-NC...[目的]探究ENO1通过NCOA4对胶质母细胞瘤(GBM)细胞铁死亡的调节机制。[方法]qRT-PCR和Western blot检测GBM组织和正常组织中ENO1的表达。人GBM细胞系U251细胞分为对照组、si-ENO1-NC组、si-ENO1-1组、si-ENO1-2组、si-ENO1+si-NCOA4-NC组、si-ENO1+si-NCOA4组,分别转染siRNA-ENO1-NC、siRNA-ENO1、siRNA-NCOA4-NC或siRNA-NCOA4序列,qRT-PCR和Western blot验证ENO1或NCOA4的表达;Western blot检测铁蛋白重多肽1(FTH1)蛋白表达;MTT法分析U251细胞增殖能力;试剂盒检测U251细胞ROS、MDA、Fe^(2+)含量及线粒体膜电位;透射电镜观察线粒体形态;PI染色检测U251细胞死亡率。[结果]与正常组织相比,GBM组织中ENO1 mRNA和蛋白表达较高(P<0.05)。与对照组和si-ENO1-NC组相比,转染si-ENO1可降低ENO1 mRNA和蛋白表达、FTH1蛋白表达以及24、48、72 h的细胞增殖活力(24 h OD值:0.35±0.05 vs 0.48±0.09,0.35±0.05 vs 0.50±0.08;48 h OD值:0.56±0.07 vs0.98±0.13,0.56±0.07 vs 1.05±0.10;72 h OD值:0.69±0.08 vs 1.35±0.14,0.69±0.08 vs 1.38±0.11)、红/绿荧光比值(3.46±0.79 vs 11.14±1.53,3.46±0.79 vs 10.97±1.82)(P<0.05),增加ROS(7.13±0.69 vs 1.00±0.12,7.13±0.69 vs 0.97±0.16)、MDA(5.61±0.53 vs 1.85±0.26,5.61±0.53 vs 1.83±0.42)、Fe^(2+)含量(6.79±0.78 vs 2.41±0.32,6.79±0.78 vs 2.46±0.47)、细胞死亡率(25.48±3.17 vs 7.31±0.84,25.48±3.17 vs 7.24±1.02)、NCOA4 mRNA和蛋白表达(P均<0.05),si-ENO1的上述作用均可被si-NCOA4逆转。[结论]ENO1缺失可上调NCOA4表达,降低FTH1蛋白水平,进而促进ROS、MDA及铁释放,诱导GBM细胞铁死亡。展开更多
目的:研究长链非编码RNA铁蛋白重链1假基因3(ferritin heavy chain 1 pseudogene 3,FTH1P3)对人肺癌顺铂(cisplatin,DDP)耐药细胞株(A549/DDP)DDP化疗敏感性的影响,并探究其潜在的分子机制。方法:采用荧光定量PCR检测FTH1P3与miR-218在A...目的:研究长链非编码RNA铁蛋白重链1假基因3(ferritin heavy chain 1 pseudogene 3,FTH1P3)对人肺癌顺铂(cisplatin,DDP)耐药细胞株(A549/DDP)DDP化疗敏感性的影响,并探究其潜在的分子机制。方法:采用荧光定量PCR检测FTH1P3与miR-218在A549和A549/DDP细胞中的表达。采用MTT法、流式细胞术、Hoechst 33258染色及CASP3活性试验评估细胞对DDP化疗敏感性的影响。采用荧光素酶报告基因和RNA蛋白免疫沉淀试验分析FTH1P3对miR-218的调控作用。结果:与A549细胞相比,A549/DDP细胞中FTH1P3表达上调,而miR-218表达下调(P<0.05)。与si-NC相比,si-FTH1P3增加A549/DDP细胞对DDP的化疗敏感性,表现为半抑制浓度(IC_(50))降低,多药耐药关联蛋白1(multidrug resistance-associated protein 1,MRP1)和ATP结合家族亚家族B成员1(ATP binding cassette subfamily B member 1,ABCB1)mRNA表达下调,细胞生长能力降低、凋亡增加及CASP3活性升高(P<0.05)。FTH1P3直接结合miR-218并抑制其表达(P<0.05)。与NC抑制物相比,miR-218抑制物能够逆转经si-FTH1P3处理的A549/DDP细胞对DDP的化疗敏感性(P<0.05)。结论:FTH1P3在A549/DDP细胞中表达上调,FTH1P3降低A549/DDP细胞对DDP的化疗敏感性,其机制与miR-218表达下调有关。展开更多
文摘Purpose: To investigate the ferritin distribution in epithelial ovarian cancer patients according to the FIGO stage in the prognosis of epithelial ovarian cancer. Method: All ovarian cancer patients were divided into two groups according their FIGO stage. Benign ovarian tumor patients were analyzed as the control. Serum ferritin, serum iron, and other related medical index were detected by automatic instruments for all patients. In addition, ferritin heavy chain (FHC) and ferritin light chain (FLC) proteins were detected by immunohistochemical staining in 60 epithelial ovarian cancer (EOC) patients and 30 benign ovarian tumor (BOT) patients, which were diagnosed in our department between 2011 and 2016. Results: The serum ferritin concentration was significantly higher in the EOC group than in the BOT group (172.56 ± 99.39 ng/mL vs 78.18 ± 43.06 ng/mL;p μmol/L vs 14.92 ± 6.36 μmol/L;p p p p p p p > 0.05). Conclusion: Patients showed an overexpression of ferritin and a downregulation of serum iron correlated with the prognosis of epithelial ovarian cancer, which may be a new target for diagnosis and treatment of epithelial ovarian cancer.
文摘目的研究四川白鹅铁蛋白重链(Ferritin heavy chain,FHC)基因编码区序列特征及其组织表达差异。方法应用分子生物学技术克隆FHC编码区序列并分析预测其编码蛋白质的结构和功能,实时荧光定量PCR技术定量检测四川白鹅13种组织中FHC的表达量。结果四川白鹅FHC编码区序列为549 bp,编码182个氨基酸,其蛋白质分子质量为21345.9,理论pI值为5.74。四川白鹅FHC氨基酸序列与北京鸭和原鸡的相似性最高(分别为100%和97%),与北京鸭具有最近的亲缘关系。FHC是稳定的可溶性蛋白质,含有高度保守的2个铁蛋白铁离子结合结构域标签和7个亚铁氧化酶双铁离子结合位点。在所检测的四川白鹅13种组织中,FHC基因均有表达,并且不同组织中FHC表达量存在明显差异。四川白鹅脾脏和视网膜中FHC的表达量显著高于其他各组织(P<0.05),肝脏、垂体和肾脏次之,肌肉中FHC表达量最低。结论四川白鹅FHC是高度保守的、组织特异性表达的基因,具有维持铁稳态和参与抗氧化的双重功能,这为阐明鹅FHC功能和铁代谢机制的研究奠定了理论基础。
文摘[目的]探究ENO1通过NCOA4对胶质母细胞瘤(GBM)细胞铁死亡的调节机制。[方法]qRT-PCR和Western blot检测GBM组织和正常组织中ENO1的表达。人GBM细胞系U251细胞分为对照组、si-ENO1-NC组、si-ENO1-1组、si-ENO1-2组、si-ENO1+si-NCOA4-NC组、si-ENO1+si-NCOA4组,分别转染siRNA-ENO1-NC、siRNA-ENO1、siRNA-NCOA4-NC或siRNA-NCOA4序列,qRT-PCR和Western blot验证ENO1或NCOA4的表达;Western blot检测铁蛋白重多肽1(FTH1)蛋白表达;MTT法分析U251细胞增殖能力;试剂盒检测U251细胞ROS、MDA、Fe^(2+)含量及线粒体膜电位;透射电镜观察线粒体形态;PI染色检测U251细胞死亡率。[结果]与正常组织相比,GBM组织中ENO1 mRNA和蛋白表达较高(P<0.05)。与对照组和si-ENO1-NC组相比,转染si-ENO1可降低ENO1 mRNA和蛋白表达、FTH1蛋白表达以及24、48、72 h的细胞增殖活力(24 h OD值:0.35±0.05 vs 0.48±0.09,0.35±0.05 vs 0.50±0.08;48 h OD值:0.56±0.07 vs0.98±0.13,0.56±0.07 vs 1.05±0.10;72 h OD值:0.69±0.08 vs 1.35±0.14,0.69±0.08 vs 1.38±0.11)、红/绿荧光比值(3.46±0.79 vs 11.14±1.53,3.46±0.79 vs 10.97±1.82)(P<0.05),增加ROS(7.13±0.69 vs 1.00±0.12,7.13±0.69 vs 0.97±0.16)、MDA(5.61±0.53 vs 1.85±0.26,5.61±0.53 vs 1.83±0.42)、Fe^(2+)含量(6.79±0.78 vs 2.41±0.32,6.79±0.78 vs 2.46±0.47)、细胞死亡率(25.48±3.17 vs 7.31±0.84,25.48±3.17 vs 7.24±1.02)、NCOA4 mRNA和蛋白表达(P均<0.05),si-ENO1的上述作用均可被si-NCOA4逆转。[结论]ENO1缺失可上调NCOA4表达,降低FTH1蛋白水平,进而促进ROS、MDA及铁释放,诱导GBM细胞铁死亡。
文摘目的:研究长链非编码RNA铁蛋白重链1假基因3(ferritin heavy chain 1 pseudogene 3,FTH1P3)对人肺癌顺铂(cisplatin,DDP)耐药细胞株(A549/DDP)DDP化疗敏感性的影响,并探究其潜在的分子机制。方法:采用荧光定量PCR检测FTH1P3与miR-218在A549和A549/DDP细胞中的表达。采用MTT法、流式细胞术、Hoechst 33258染色及CASP3活性试验评估细胞对DDP化疗敏感性的影响。采用荧光素酶报告基因和RNA蛋白免疫沉淀试验分析FTH1P3对miR-218的调控作用。结果:与A549细胞相比,A549/DDP细胞中FTH1P3表达上调,而miR-218表达下调(P<0.05)。与si-NC相比,si-FTH1P3增加A549/DDP细胞对DDP的化疗敏感性,表现为半抑制浓度(IC_(50))降低,多药耐药关联蛋白1(multidrug resistance-associated protein 1,MRP1)和ATP结合家族亚家族B成员1(ATP binding cassette subfamily B member 1,ABCB1)mRNA表达下调,细胞生长能力降低、凋亡增加及CASP3活性升高(P<0.05)。FTH1P3直接结合miR-218并抑制其表达(P<0.05)。与NC抑制物相比,miR-218抑制物能够逆转经si-FTH1P3处理的A549/DDP细胞对DDP的化疗敏感性(P<0.05)。结论:FTH1P3在A549/DDP细胞中表达上调,FTH1P3降低A549/DDP细胞对DDP的化疗敏感性,其机制与miR-218表达下调有关。
