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Adiponectin receptor 1 and small ubiquitin-like modifier 4 polymorphisms are associated with risk of coronary artery disease without diabetes 被引量:4
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作者 Hong LI Ze YANG +9 位作者 Lian-Mei PU Xiang LI Yang RUAN Fan YANG Shuai MENG Duo YANG Wei YAO Hao FU Feng ZHANG Ze-Ning JIN 《Journal of Geriatric Cardiology》 SCIE CAS CSCD 2016年第9期776-782,共7页
Background The genes encoding adiponectin receptor 1 (ADIPOR1) and small ubiquitin-like modifier 4 (SUM04) have been linked to anti-atherogenic effects, but little is known about whether polymorphisms in the two g... Background The genes encoding adiponectin receptor 1 (ADIPOR1) and small ubiquitin-like modifier 4 (SUM04) have been linked to anti-atherogenic effects, but little is known about whether polymorphisms in the two genes, acting separately or interacting, affect risk of coronary artery disease (CAD) without diabetes. Methods We genotyped 200 CAD patients without diabetes and 200 controls without CAD or diabetes at three single-nucleotide polymorphisms (SNPs) in ADIPOR1 and one SNP in SUM04, which were chosen based on previous studies. Potential associations were also explored between these SNPs and clinical characteristics of CAD without diabetes. Results Risk alleles at three SNPs inADIPOR1 (rs7539542-G, rs7514221-C and rs3737884-G) and the G allele at SNP rs237025 in SUM04 significantly increased risk of CAD without diabetes, with ORs ranging from 1.79 to 4.44. Carriers of any of these four risk alleles showed similar adverse clinical characteristics. Compared with individuals with a CC or GC genotype, those with a GG genotype at rs3737884 were at significantly higher risk of CAD that affected the left anterior descending coronary artery (OR: 6.77, P = 0.009), the right coronary artery (OR: 4.81, P = 0.028) or a relatively large number of vessels (P = 0.04). Individuals carrying a risk allele at one or more of the three SNPs in ADIPOR1 as well as a risk allele at the SNP in SUM04 were at significantly higher risk of CAD without diabetes than individuals not carrying any risk alleles (OR: 5.82, 95% CI: 1.23-27.7, P= 0.013). Conelusions SNPs in ADIPORl and SUMO4 are associated with elevated risk of CAD without diabetes, and SNPs in the two genes may interact to jointly affect disease risk. 展开更多
关键词 Adiponectin receptor 1 Coronary artery disease DIABETES POLYMORPHISM small ubiquitin-like modifier 4
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布鲁氏菌侵染对类泛素SUMO-1表达的影响及类泛素SUMO-1慢病毒表达载体的构建 被引量:1
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作者 李启峰 丁金花 +2 位作者 窦文静 张辉 陈创夫 《中国畜牧兽医》 CAS 北大核心 2016年第6期1422-1429,共8页
研究布鲁氏菌侵染小鼠巨噬细胞RAW264.7后类泛素SUMO-1的表达变化,并构建小鼠类泛素SUMO-1基因过表达的慢病毒载体。本试验分别利用布鲁氏菌16M、M5-90侵染小鼠巨噬细胞RAW264.7,利用实时荧光定量PCR和Western blotting分别检测细胞中... 研究布鲁氏菌侵染小鼠巨噬细胞RAW264.7后类泛素SUMO-1的表达变化,并构建小鼠类泛素SUMO-1基因过表达的慢病毒载体。本试验分别利用布鲁氏菌16M、M5-90侵染小鼠巨噬细胞RAW264.7,利用实时荧光定量PCR和Western blotting分别检测细胞中类泛素SUMO-1的表达;利用DNA重组技术将类泛素SUMO-1基因片段插入到慢病毒表达载体pLEX-MCS中,获得重组慢病毒质粒pLEX-SUMO-1,测序鉴定成功后转染293T细胞,包装好的慢病毒感染小鼠巨噬细胞RAW264.7,并利用实时荧光定量PCR、Western blotting方法分别检测细胞中类泛素SUMO-1mRNA及蛋白表达水平。结果表明,布鲁氏菌16M、M5-90侵染细胞后,在感染早期类泛素SUMO-1的表达受到抑制,12h内呈现下降趋势,在12h后开始上升,极显著高于正常水平(P<0.01);测序结果证明,类泛素SUMO-1基因正确插入到pLEX-MCS质粒中;实时荧光定量PCR方法检测表明,类泛素SUMO-1mRNA转录水平上调。由此可见,布鲁氏菌侵染小鼠巨噬细胞RAW264.7能够引起细胞内类泛素SUMO-1表达的改变;成功构建了类泛素SUMO-1慢病毒过表达载体,为进一步研究类泛素SUMO-1的功能奠定基础。 展开更多
关键词 布鲁氏菌 类泛素sumo-1 慢病毒载体 小鼠巨噬细胞
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Over-expression of small ubiquitin-like modifier proteases 1 predicts chemo-sensitivity and poor survival in non-small cell lung cancer 被引量:8
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作者 Mu Juwei Zuo Yong +7 位作者 Yang Wenjing Chen Zhaoli Liu Ziyuan Tu Jun Li Yan Yuan Zuyang Cheng Jinke He Jie 《Chinese Medical Journal》 SCIE CAS CSCD 2014年第23期4060-4065,共6页
Background Non-small cell lung cancer (NSCLC) is one of the most common malignant tumors.Despite the advances in therapy over the years,its mortality remains high.The aim of this study was to evaluate the expression... Background Non-small cell lung cancer (NSCLC) is one of the most common malignant tumors.Despite the advances in therapy over the years,its mortality remains high.The aim of this study was to evaluate the expression of small ubiquitin-like modifier (SUMO) proteases 1 (SENP1) in NSCLC tissues and its role in the regulation of vascular endothelial growth factor (VEGF) expression.We also investigated the association between the expression level of SENP1 and the clinicopathological features and survival of the patients.Methods A SENP1 small interfering RNA (siRNA) was constructed and transfected into the NSCLC cells.VEGF gene expression was analyzed by real-time polymerase chain reaction (RT-PCR).Immunohistochemistry staining was used to assess the expression of SENP1 in 100 NSCLC patients and its association with the clinicopathological features and survival was analyzed.Results VEGF expression was significantly higher in NSCLC tissues than in normal lung tissues.Inhibition of SENP1 by siRNA was associated with decreased VEGF expression.SENP1 was over-expressed in 55 of the 100 NSCLC samples (55%) and was associated with a moderate and low histological tumor grade (3.6%,38.2%,and 58.2% in high,moderate and low differentiated tumors,respectively,P=0.046),higher T stage (10.9% in T1,and 89.1% in T2 and T3 tumor samples,P <0.001)and TNM stage (10.9% in stage Ⅰ,and 89.1% in stages Ⅱ and Ⅲ tumor samples,P <0.001).The rate of lymph node metastasis was significantly higher in the SENP1 over-expression group (76.4%) than that in the SENP1 low expression group (33.3%,P <0.001).Sixty three patients received postoperative chemotherapy,including 34 with SENP1 over-expression and 29 with SENP1 low expression.Among the 34 patients with SENP1 over-expression,22 (64.7%) patients developed recurrence or metastasis,significantly higher than those in the low expression group 27.6% (8/29) (P=0.005).Multivariate Cox regression analysis showed that lymph node metastasis (P=0.015),TNM stage (P=-0.001),and SENP1 expression level (P=0.002) were independent prognostic factors for the survival of NSCLC patients.Conclusions SENP1 may be a promising predictor of survival,a predictive factor of chemo-sensitivity for NSCLC patients,and potentially a desirable drug target for lung carcinoma target therapy. 展开更多
关键词 small ubiquitin-like modifier proteases 1 (SENP1 non-small cell lung cancer PROGNOSIS neoplasm recurrence
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Construction,Expression and Purification of SUMO1-GST Fusion Protein
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作者 QIAO Xiao-fang FANG Xue-dong LIU Jun 《Chemical Research in Chinese Universities》 SCIE CAS CSCD 2011年第2期245-248,共4页
Sumoylation is an important protein modification discovered recently. SUMO(small ubiquitin-related modifier) pathway regulates the protein stability and transcriptional activity with a 12-kDa small molecular protein... Sumoylation is an important protein modification discovered recently. SUMO(small ubiquitin-related modifier) pathway regulates the protein stability and transcriptional activity with a 12-kDa small molecular protein, SUMO, ligated to the target protein. The purification of SUMO proteins is a key step to reveal their function. The purpose of this study was to construct the recombinant SUMO1 gene cloned to a pGEX-4T-1 vector to express and purify the SUMO1-GST fusion protein in Escherichia coli. First, the full length DNA sequence of SUMO1 gene was amplified by PCR and was ligated to pMD18-T vector. Then the SUMO1 gene was subcloned to pGEX-4T-1 prokaryotic expression vector between BamHI and XhoI sites, and transformed in Escherichia coli DH5α cells. The right colonies were identified by restrictive enzyme digestion and sequencing. The correct rebombinant plasmid of pGEX-4T-1-SUMO1 was transformed in Escherichia coli BL21 cells and then induced by IPTG(isopropyl- β-D-1- thiogalacto-pyranoside) to express the SUMO1-GST fusion protein. The highly purified SUMO1-GST(glutathione S-transferase) fusion protein was obtained by affinity chromatography. Finally, the properties of SUMO1-GST fusion protein were confirmed by Coomassie brilliant blue strain and Western blot analysis. The recombinant plasmid of pGEX-4T-1-SUMO1 was successfully constructed, and SUMO1-GST fusion proteins were successfully expressed. 展开更多
关键词 small ubiquitin-related modifier 1 sumo1 Gutathione S-transferase(GST) fusion protein Affinity chromatography
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SUMO特异性蛋白酶1诱导蛋白质去小泛素相关修饰增加子宫内膜癌侧群细胞化疗敏感性 被引量:1
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作者 袁梦岚 白洁 +5 位作者 李承尧 薛娜 陈旭红 盛凤 刘晓智 李圃 《中华肿瘤杂志》 CAS CSCD 北大核心 2022年第12期1362-1368,共7页
目的探讨通过诱导蛋白质广泛去类泛素化修饰途径抑制子宫内膜癌的干性维持潜能,达到子宫内膜癌侧群细胞化疗增敏的目的。方法流式细胞术分选培养CD133^(+)CD44^(+)的KLE子宫内膜癌侧群细胞克隆球,Western blot法检测小泛素相关修饰蛋白1... 目的探讨通过诱导蛋白质广泛去类泛素化修饰途径抑制子宫内膜癌的干性维持潜能,达到子宫内膜癌侧群细胞化疗增敏的目的。方法流式细胞术分选培养CD133^(+)CD44^(+)的KLE子宫内膜癌侧群细胞克隆球,Western blot法检测小泛素相关修饰蛋白1(SUMO1)及肿瘤侧群细胞干性维持基因八聚体结合转录因子4(Oct4)和性别决定区Y-box2(Sox2)蛋白的表达情况。慢病毒介导KLE侧群细胞稳定转染SUMO特异性蛋白酶1(SENP1)基因,Western blot法检测SENP1、SUMO1、Oct4和Sox2的蛋白表达;比较过表达与未过表达SENP1基因的KLE侧群细胞克隆形成率,流式细胞术检测细胞周期变化,四甲基偶氮唑蓝实验和流式细胞术检测子宫内膜癌侧群细胞对顺铂的化疗敏感性,子宫内膜癌荷瘤鼠模型检测SENP1过表达对顺铂的化疗敏感性影响。结果CD133^(+)CD44^(+)和CD133-CD44-子宫内膜癌KLE细胞克隆形成率分别为(23.64±5.03)%和(4.64±1.25)%,差异有统计学意义(P=0.003),CD133^(+)CD44^(+)子宫内膜癌KLE细胞中SUMO1、Oct4和Sox2蛋白表达水平高于CD133-CD44-子宫内膜癌KLE细胞(均P<0.05)。与未转染SENP1基因的KLE侧群细胞比较,SENP1过表达的KLE侧群细胞SUMO1、Oct4、Sox2蛋白表达水平降低,细胞克隆形成率由(25.67±5.44)%下降至(7.46±1.42)%,细胞周期发生由G0/G_(1)期向G_(2)期的偏移,顺铂半数抑制浓度由(55.46±6.14)μg/ml下降至(11.55±3.12)μg/ml,细胞凋亡率由(9.76±2.09)%上升至(16.79±3.44)%,差异均有统计学意义(均P<0.05)。过表达SENP1能够降低KLE侧群细胞增殖能力,增加化疗敏感性。结论通过过表达SENP1能够诱导蛋白质去SUMO化修饰,抑制子宫内膜癌侧群细胞的干性维持潜能,进而增强其化疗敏感性。 展开更多
关键词 子宫内膜肿瘤 小泛素相关修饰蛋白 sumo特异性蛋白酶1 肿瘤侧群细胞 顺铂
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结直肠癌组织泛素样小分子修饰因子-1表达及其临床意义 被引量:2
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作者 麦蕾 赖人旭 +1 位作者 林宇静 郭惠学 《中华肿瘤防治杂志》 CAS 北大核心 2015年第2期113-115,119,共4页
目的探讨泛素样小分子修饰因子-1(small ubiquitin-like modifier-1,SUMO-1)蛋白在结直肠癌组织中的表达及其临床意义。方法收集2010-01-01-2012-09-30在中山大学附属第五医院手术切除或结肠镜下切除的标本,其中63例结直肠癌组织,47例... 目的探讨泛素样小分子修饰因子-1(small ubiquitin-like modifier-1,SUMO-1)蛋白在结直肠癌组织中的表达及其临床意义。方法收集2010-01-01-2012-09-30在中山大学附属第五医院手术切除或结肠镜下切除的标本,其中63例结直肠癌组织,47例结直肠腺瘤组织,10例正常结直肠黏膜或炎症组织。采用免疫组织化学SP法检测各组织中SUMO-1蛋白的表达情况。分析SUMO-1蛋白与结直肠癌患者不同临床病理因素之间的关系。应用SPSS 13.0进行统计学分析,计数资料比较用χ2检验和χ2检验连续性校正法,等级资料比较用秩和检验。结果 SUMO-1蛋白在结直肠癌组织的阳性表达率为95.2%(60/63),结直肠腺瘤组织为21.3%(10/47),正常结直肠黏膜或炎症组织中为10.0%(1/10)。其在结直肠癌组织与结直肠腺瘤组织的表达差异有统计学意义,χ2=63.633,P<0.001;在结直肠癌组织与正常结直肠黏膜或炎症组织中的表达差异有统计学意义,χ2=39.653,P<0.001;在结直肠腺瘤组织与正常结直肠黏膜或炎症组织中的表达差异无统计学意义,χ2=0.144,P=0.704。SUMO-1蛋白的表达与患者的性别、远处转移、TNM分期、浸润深度和组织学分级明显相关,P值均<0.05。但与患者年龄、肿瘤大小、大体类型和淋巴结转移无关,P值均>0.05。结论 SUMO-1蛋白在结直肠癌组织中的表达明显增高。SUMO-1蛋白的表达与癌组织的浸润深度、TNM分期和远处转移相关。 展开更多
关键词 结直肠肿瘤 泛素样小分子修饰因子-1 sumo-1 免疫组织化学
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新型重组Ulp1的制备工艺研究
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作者 徐海悦 吴勇 +3 位作者 李乐乐 黄宗庆 化浩举 冯军 《中国医药工业杂志》 CAS CSCD 北大核心 2022年第10期1432-1438,共7页
类泛素特异性蛋白酶1(Ulp1)是从融合蛋白中移除小分子泛素相关修饰物蛋白(SUMO)标签工艺中的重要工具蛋白酶,在重组多肽和蛋白生产制备中具有广泛应用。Ulp1蛋白易聚集的特性给下游分离纯化工艺带来了诸多挑战。为克服这些问题,本研究... 类泛素特异性蛋白酶1(Ulp1)是从融合蛋白中移除小分子泛素相关修饰物蛋白(SUMO)标签工艺中的重要工具蛋白酶,在重组多肽和蛋白生产制备中具有广泛应用。Ulp1蛋白易聚集的特性给下游分离纯化工艺带来了诸多挑战。为克服这些问题,本研究设计并重组表达制备了一种S13-Ulp1融合蛋白,以提高Ulp1酶的溶解度,减少聚集,同时降低蛋白等电点,便于采用阴离子交换色谱纯化。通过对发酵、澄清等步骤的工艺参数进行优化,建立了一条发酵表达产量高、纯化工艺简便的S13-Ulp1融合蛋白制备工艺。优化后发酵产量达到了2.34g/L,下游分离纯化总收率为64%,制得的S13-Ulp1蛋白的SDS-PAGE纯度为95%。经酶切试验验证,S13-Ulp1融合蛋白具有与Ulp1相同的特异性,可高效地移除SUMO标签,获得目的多肽。 展开更多
关键词 小分子泛素相关修饰物蛋白(sumo)蛋白酶:类泛素特异性蛋白酶1(Ulp1) 重组 表达 纯化 离子交换色谱
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