Bloom综合症(Bloom s syndrome)是人类一种少见的常染色体隐性遗传疾病,BLM基因突变导致这种疾病的发生,患者染色体极不稳定,是多种癌症的易患体,其致病机理不清楚.该研究在优化诱导表达温度、时间、IPTG质量浓度、pH值、培养基种类以...Bloom综合症(Bloom s syndrome)是人类一种少见的常染色体隐性遗传疾病,BLM基因突变导致这种疾病的发生,患者染色体极不稳定,是多种癌症的易患体,其致病机理不清楚.该研究在优化诱导表达温度、时间、IPTG质量浓度、pH值、培养基种类以及培养基成分的基础上,建立了BLM642-1290重组蛋白表达、分离和纯化方法.最优表达条件为IPTG0.45 mmol/L,诱导温度18℃,诱导表达时间20 h,pH值7.0.在LB培养基中添加终质量浓度为5 mmol/L的EDTA能够增加蛋白的表达量.该实验所建立的方法为进一步开展Bloom综合症的研究奠定了基础.展开更多
AIM:To investigate the biological role of mi R-1290 in esophageal squamous cell carcinoma(ESCC) progression and invasion and the underlying mechanism.METHODS:Quantitative real-time polymerase chain reaction(q RT-PCR) ...AIM:To investigate the biological role of mi R-1290 in esophageal squamous cell carcinoma(ESCC) progression and invasion and the underlying mechanism.METHODS:Quantitative real-time polymerase chain reaction(q RT-PCR) was performed to evaluate mi R-1290 expression in ESCC tissue samples.The roles of mi R-1290 in cell proliferation,migration and invasion were identified using mi R-1290 mimic-transfected cells.In addition,the regulatory effect of mi R-1290 on suppressor of cancer cell invasion(SCAI) was evaluated using q RT-PCR,Western blot analysis and a dual luciferase reporter assay.RESULTS:mi R-1290 was significantly upregulated in ESCC tissue samples compared with normal adjacent tissues(9.213 ± 1.150 vs 1.000 ± 0.0),(P < 0.01).Upregulation of mi R-1290 was associated with tumor differentiation(P = 0.021),N classification(P = 0.006) and tumor-node-metastasis stage(P = 0.021) in ESCC patients.Moreover,ectopic mi R-1290 expression potently promoted ESCC cell growth(P < 0.01),migration(P < 0.01) and invasion(P < 0.01) in vitro.mi R-1290 overexpression in ESCC cell lines decreased SCAI expression at the translational level and reduced SCAI-driven luciferase-reporter activity(P < 0.01).CONCLUSION:Our findings suggested that mi R-1290 may play an oncogenic role in cellular processes of ESCC.展开更多
Objective Bloom’s syndrome is an autosomal recessive disorder characterized by genomic instability and a predisposition to many cancers. Mutations of the BLM gene （encoding a BLM helicase） may form a structure of t...Objective Bloom’s syndrome is an autosomal recessive disorder characterized by genomic instability and a predisposition to many cancers. Mutations of the BLM gene （encoding a BLM helicase） may form a structure of the etiology of this disease. As a global pollutant, mercury poses a major threat to human health. The current study was conducted to elucidate the effects of Hg^2＋ on the structure and activity of BLM642‐1290 recombinant helicase, and to further explore the molecular mechanisms of mercury toxicity to the DNA helicase. Methods The effects of Hg^2＋ on biological activity and structure of BLM642‐1290 recombinant helicase were determined by fluorescence polarized, ultraviolet spectroscopic, and free‐phosphorus assay technologies, respectively. Results The helicase activity, the DNA‐binding activity, and the ATPase activity of BLM642‐1290 recombinant helicase were inhibited by Hg^2＋ treatment. The LMCT （ligand‐to‐metal charge transition） peaks of the helicase were enhanced with the increase of the Hg^2＋ level. The LMCT peaks of the same concentration of helicase gradually increased over time. Conclusions The biological activity of BLM642‐1290 recombinant helicase is inhibited by Hg^2＋ treatment. The conformation of the helicase is significantly altered by Hg^2＋ . There exist two binding sites between Hg^2＋ and the helicase, which are located in the amino acid residues 1063‐1066 and 940‐944 of the helicase, respectively.展开更多
目的:探究胃癌患者血浆miR-1290的表达及其临床意义。方法:收集2017年10月至2018年6月间就诊于安徽省立医院的84例胃癌患者的血浆及临床病理资料,所有患者均经胃镜及病理活检确诊且术前未进行放疗、化疗或其他药物治疗,同时收集年龄、...目的:探究胃癌患者血浆miR-1290的表达及其临床意义。方法:收集2017年10月至2018年6月间就诊于安徽省立医院的84例胃癌患者的血浆及临床病理资料,所有患者均经胃镜及病理活检确诊且术前未进行放疗、化疗或其他药物治疗,同时收集年龄、性别与之相匹配的健康者血浆29例,检测血浆中miR-1290的表达水平,并分析胃癌患者血浆miR-1290表达量与临床病理参数之间的关系。利用受试者工作曲线评估血浆miR-1290作为生物标志物在诊断胃癌方面的临床价值。结果:胃癌患者与健康对照组相比,血浆miR-1290的表达量明显增高(6.44±2.59 vs 3.44±1.75),差异有统计学意义(P<0.001)。miR-1290在胃癌患者血浆中的表达水平与患者的年龄、性别无相关性,但与肿瘤的大小、肿瘤的浸润深度、淋巴结转移、TNM分期有显著相关性。ROC曲线结果表明miR-1290对胃癌的发生有预测作用,最佳截断值为3.5,此时灵敏度为86.9%,特异度为65.6%。结论:miR-1290在胃癌患者血浆中表达上调,其表达水平与胃癌的发生、发展密切相关,有望成为胃癌诊断的指标。展开更多
基金Supported by grants from Innovative Team Project of Jiangsu Province,China,No.CXZZ11_0705
文摘AIM:To investigate the biological role of mi R-1290 in esophageal squamous cell carcinoma(ESCC) progression and invasion and the underlying mechanism.METHODS:Quantitative real-time polymerase chain reaction(q RT-PCR) was performed to evaluate mi R-1290 expression in ESCC tissue samples.The roles of mi R-1290 in cell proliferation,migration and invasion were identified using mi R-1290 mimic-transfected cells.In addition,the regulatory effect of mi R-1290 on suppressor of cancer cell invasion(SCAI) was evaluated using q RT-PCR,Western blot analysis and a dual luciferase reporter assay.RESULTS:mi R-1290 was significantly upregulated in ESCC tissue samples compared with normal adjacent tissues(9.213 ± 1.150 vs 1.000 ± 0.0),(P < 0.01).Upregulation of mi R-1290 was associated with tumor differentiation(P = 0.021),N classification(P = 0.006) and tumor-node-metastasis stage(P = 0.021) in ESCC patients.Moreover,ectopic mi R-1290 expression potently promoted ESCC cell growth(P < 0.01),migration(P < 0.01) and invasion(P < 0.01) in vitro.mi R-1290 overexpression in ESCC cell lines decreased SCAI expression at the translational level and reduced SCAI-driven luciferase-reporter activity(P < 0.01).CONCLUSION:Our findings suggested that mi R-1290 may play an oncogenic role in cellular processes of ESCC.
基金supported by the Natural Sciences Foundation of China (NSFC, No.30660043)the National Basic Research Program of China (2010CB534912)+1 种基金the Doctoral Program of the Ministry of Education of China (No.200806570003)the Governor Talents Foundation of Guizhou Province (No.200822)
文摘Objective Bloom’s syndrome is an autosomal recessive disorder characterized by genomic instability and a predisposition to many cancers. Mutations of the BLM gene （encoding a BLM helicase） may form a structure of the etiology of this disease. As a global pollutant, mercury poses a major threat to human health. The current study was conducted to elucidate the effects of Hg^2＋ on the structure and activity of BLM642‐1290 recombinant helicase, and to further explore the molecular mechanisms of mercury toxicity to the DNA helicase. Methods The effects of Hg^2＋ on biological activity and structure of BLM642‐1290 recombinant helicase were determined by fluorescence polarized, ultraviolet spectroscopic, and free‐phosphorus assay technologies, respectively. Results The helicase activity, the DNA‐binding activity, and the ATPase activity of BLM642‐1290 recombinant helicase were inhibited by Hg^2＋ treatment. The LMCT （ligand‐to‐metal charge transition） peaks of the helicase were enhanced with the increase of the Hg^2＋ level. The LMCT peaks of the same concentration of helicase gradually increased over time. Conclusions The biological activity of BLM642‐1290 recombinant helicase is inhibited by Hg^2＋ treatment. The conformation of the helicase is significantly altered by Hg^2＋ . There exist two binding sites between Hg^2＋ and the helicase, which are located in the amino acid residues 1063‐1066 and 940‐944 of the helicase, respectively.
文摘目的:探究胃癌患者血浆miR-1290的表达及其临床意义。方法:收集2017年10月至2018年6月间就诊于安徽省立医院的84例胃癌患者的血浆及临床病理资料,所有患者均经胃镜及病理活检确诊且术前未进行放疗、化疗或其他药物治疗,同时收集年龄、性别与之相匹配的健康者血浆29例,检测血浆中miR-1290的表达水平,并分析胃癌患者血浆miR-1290表达量与临床病理参数之间的关系。利用受试者工作曲线评估血浆miR-1290作为生物标志物在诊断胃癌方面的临床价值。结果:胃癌患者与健康对照组相比,血浆miR-1290的表达量明显增高(6.44±2.59 vs 3.44±1.75),差异有统计学意义(P<0.001)。miR-1290在胃癌患者血浆中的表达水平与患者的年龄、性别无相关性,但与肿瘤的大小、肿瘤的浸润深度、淋巴结转移、TNM分期有显著相关性。ROC曲线结果表明miR-1290对胃癌的发生有预测作用,最佳截断值为3.5,此时灵敏度为86.9%,特异度为65.6%。结论:miR-1290在胃癌患者血浆中表达上调,其表达水平与胃癌的发生、发展密切相关,有望成为胃癌诊断的指标。
