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XCAP-C-like Protein Existing in Nuclei and Chromosomes of Allium sativa
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作者 王岩 邢苗 阎石 《Acta Botanica Sinica》 CSCD 2002年第1期29-33,共5页
In studying the XCAP-C-like protein in the root meristematic cells of Allium sativa L., the nuclei were isolated from the cells and the nuclear matrices prepared. A 165 kD polypeptide, which is equivalent to XCAP-C in... In studying the XCAP-C-like protein in the root meristematic cells of Allium sativa L., the nuclei were isolated from the cells and the nuclear matrices prepared. A 165 kD polypeptide, which is equivalent to XCAP-C in molecular weight, was demonstrated in the nuclei by SDS-PAGE, and was then proved to be an XCAP-C-like protein by Western blot using an anti-XCAP-C antiserum, but neither the polypeptide nor the XCAP-C-like protein was detected in die nuclear matrix. The nuclei, Chromosomes and chromosome scaffolds were observed to emanate strong, specific fluorescence after labeled with the anti-XCAP-C antiserum and an FITC-conjugated secondary antibody, indicating their containment of the XCAP-C-like protein. It was confirmed by viewing with immunoelectron microscopy that the gold particles representing the localization of the XCAP-C-like protein were found to be mainly distributed in the condensed chromatin regions of the nuclei and chromosomes. 展开更多
关键词 Allium saliva SMC protein XCAP-C-like protein nucleus chromosome
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Transfer of disulfide bond formation modules via yeast artificial chromosomes promotes the expression of heterologous proteins in Kluyveromyces marxianus
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作者 Pingping Wu Wenjuan Mo +6 位作者 Tian Tian Kunfeng Song Yilin Lyu Haiyan Ren Jungang Zhou Yao Yu Hong Lu 《mLife》 CSCD 2024年第1期129-142,共14页
Kluyveromyces marxianus is a food-safe yeast with great potential for producing heterologous proteins.Improving the yield in K.marxianus remains a challenge and incorporating large-scale functional modules poses a tec... Kluyveromyces marxianus is a food-safe yeast with great potential for producing heterologous proteins.Improving the yield in K.marxianus remains a challenge and incorporating large-scale functional modules poses a technical obstacle in engineering.To address these issues,linear and circular yeast artificial chromosomes of K.marxianus(KmYACs)were constructed and loaded with disulfide bond formation modules from Pichia pastoris or K.marxianus.These modules contained up to seven genes with a maximum size of 15 kb.KmYACs carried telomeres either from K.marxianus or Tetrahymena.KmYACs were transferred successfully into K.marxianus and stably propagated without affecting the normal growth of the host,regardless of the type of telomeres and configurations of KmYACs.KmYACs increased the overall expression levels of disulfide bond formation genes and significantly enhanced the yield of various heterologous proteins.In high-density fermentation,the use of KmYACs resulted in a glucoamylase yield of 16.8 g/l,the highest reported level to date in K.marxianus.Transcriptomic and metabolomic analysis of cells containing KmYACs suggested increased flavin adenine dinucleotide biosynthesis,enhanced flux entering the tricarboxylic acid cycle,and a preferred demand for lysine and arginine as features of cells overexpressing heterologous proteins.Consistently,supplementing lysine or arginine further improved the yield.Therefore,KmYAC provides a powerful platform for manipulating large modules with enormous potential for industrial applications and fundamental research.Transferring the disulfide bond formation module via YACs proves to be an efficient strategy for improving the yield of heterologous proteins,and this strategy may be applied to optimize other microbial cell factories. 展开更多
关键词 disulfide bond formation expression of heterologous proteins Kluyveromyces marxianus TELOMERE yeast artificial chromosome
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Human chromosome pellicle antibody recognizing centromere protein-C(CENP-C),the main component of the kinetochore
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作者 XIE YONG ZU MEI NI +3 位作者 JIAN REN GU PHIL WONG WEN QING WU GUO WEI XU(Hong Kong University of Science and Technology,Department of Biology, Hong Kong) (Shanghai Institute of Cell Biology, Chinese Academy of Sciences, Shanghai) (Shanghai Cancer Institute, Nation 《Cell Research》 SCIE CAS CSCD 1997年第1期13-19,共7页
Recently the antichromosome antisera from several scleroderma patients have been found to recognize the pellicle of metaphase and anaphase chromosomes. In order to identify the pellicle components, we used these antic... Recently the antichromosome antisera from several scleroderma patients have been found to recognize the pellicle of metaphase and anaphase chromosomes. In order to identify the pellicle components, we used these antichromosome antisera to screen a human embryonic cDNA library. The sequences of the positive clones are identical to the cDNA gene sequence of CENP-C (centromere protein C), a human centromere autoantigen. This result suggusts that CENP-C is a component of the pellicle of human metaphase and anaphase chromosomes. 展开更多
关键词 Human antibody scleroderma CENP-C (centromere protein C) METAPHASE chromosome pellicle indirect immunofluorescent staining
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C-type lectins and human epithelial membrane protein1:Are they new proteins in keratin disorders?
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作者 Nilufer Karadeniz Thomas Liehr +4 位作者 Kristin Mrasek Ibrahim Asik Zuleyha Asik Nadezda Kosyakova Hasmik Mkrtchyan 《Open Journal of Genetics》 2013年第4期262-269,共8页
Here we report a family with a clinical spectrum of Pachyonychia Congenita Tarda (PCT) encompassing two generations via a balanced chromosomal translocation between 4q26 and 12p12.3. We discuss the effects of chromoso... Here we report a family with a clinical spectrum of Pachyonychia Congenita Tarda (PCT) encompassing two generations via a balanced chromosomal translocation between 4q26 and 12p12.3. We discuss the effects of chromosomal translocations on gene expression through involved breakpoints and structural gene abnormalities detected by array CGH. We believe that the family we present gives further insight to the better understanding of molecular and structural basis of keratin disorders, and to the late onset and genetic basis of PCT through the possible role of C-type lectins and human epithelial membrane protein1 (EMP1). Better understanding of the molecular basis of keratin disorders is the foundation for improved diagnosis, genetic counseling and novel therapeutic approaches to overcome the current treatment limitations related to this disease. 展开更多
关键词 KERATINS Palmoplantar Keratoderma Pachyonychia Congenital Tarda LECTINS Epithelial Membrane protein1 Gene Expression and chromosome Translocation
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Phosphatase and tensin homology deleted in chromosome 10,hypoxia-inducible factor-1 alpha gene expression in colorectal adenoma and adenocarcinoma and their relation to vascular endothelial growth factor protein expression
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作者 钱群 《外科研究与新技术》 2005年第3期165-166,共2页
To examine phosphatase and tensin homology deleted in chromosome 10 (PTEN),hypoxia-inducible factor-1 alpha (HIF-1 alpha) gene expressions and their relation to vascular endothelial growth factor(VEGF) protein express... To examine phosphatase and tensin homology deleted in chromosome 10 (PTEN),hypoxia-inducible factor-1 alpha (HIF-1 alpha) gene expressions and their relation to vascular endothelial growth factor(VEGF) protein expression in the patients with human colorectal adenomas and adenocarcinomas.Methods The expression of PTEN,HIF-1 alpha gene was detected by using in situ hybridization,and the VEGF expression levels by immunohistochemistry in colorectal adenomas and primary colorectal adenocarcinoma.Results Strong expression of HIF-1 alpha was detectable in the majority of colorectal dadenocarcinoma,particularly surrounding areas of necrosis in adenocarcinoma.PTEN,HIF-1 alpha mRNA and VEGF protein were positive in 51.6%,67.7% and 59.7% respectively in 62 cases of adenocarcinomas,and 77.8%,44.4% and 33.3% respectively in 18 cases of adenomas.The positive rate of VEGF was higher in the patients with colorectal adenocarcinomas than that in those with adenomas,whereas that of PTEN mRNA was contrary.HIF-1 mRNA expression was correlated significantly with lymph node metastasis,liver metastasis,Duke’s stage and recurrence.During colorectal tumor progression,the expression of HIF-1 alpha mRNA was positively correlated with the VEGF protein expression (χ2= 4.751 ,P<0.05),but negatively with the PTEN mRNA expression(χ2=21.84,P<0.01).Conclusion The absence or low expression of PTEN and the increased levels of HIF-1α and VEGF may paly an important role in carcinogenesis and progression of colorectal carcinoma.These results suggest that VEGF upregulated by HIF-1 alpha gene may be involved in angiogenesis of colorectal adenocarcinoma.4 refs,1 tab. 展开更多
关键词 Phosphatase and tensin homology deleted in chromosome 10 hypoxia-inducible factor-1 alpha gene expression in colorectal adenoma and adenocarcinoma and their relation to vascular endothelial growth factor protein expression
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河北省免费NIPT联合PAPP-A、F-β-HCG 及超声检查诊断孕中期胎儿染色体异常的意义
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作者 李丽霞 朱晓明 韩文莹 《中国实验诊断学》 2024年第4期439-443,共5页
目的 评价河北行免费的无创产前DNA检测(NIPT),联合血清妊娠相关血浆蛋白-A(PAPP-A)、游离β-人绒毛膜促性腺激素(F-β-HCG)检测及超声对孕中期染色体异常的诊断价值。方法 选取衡水市第二人民医院2019年1月至2023年1月接收的3 591例拟... 目的 评价河北行免费的无创产前DNA检测(NIPT),联合血清妊娠相关血浆蛋白-A(PAPP-A)、游离β-人绒毛膜促性腺激素(F-β-HCG)检测及超声对孕中期染色体异常的诊断价值。方法 选取衡水市第二人民医院2019年1月至2023年1月接收的3 591例拟行染色体检测的孕中期妇女,均行免费NIPT联合血清学指标(PAPP-A、F-β-HCG)及超声检查。将羊水穿刺染色体核型分析结果记为金标准,采用受试者工作特征曲线(ROC)评价不同方法诊断孕中期染色体异常的价值。结果 经随访,在3 591例中,共有39例染色体异常,其中,32例经超声检查有染色体异常,34例经NIPT检测有染色体异常。在3 552例染色体正常的胎儿中,34例经超声检查有染色体异常,36例经NIPT检测有染色体异常;染色体异常胎儿的母体PAPP-A、F-β-HCG水平均高于染色体正常胎儿的母体(P<0.05)。PAPP-A、F-β-HCG共同诊断胎儿染色体异常的敏感度及ROC曲线下方的面积(AUC)均高于母体PAPP-A、F-β-HCG单独诊断(P<0.05);NIPT,PAPP-A、F-β-HCG及超声的联合诊断的敏感度及AUC均高于NIPT,PAPP-A、F-β-HCG,超声的单独诊断(P<0.05),且不同方法诊断染色体异常的特异度对比差异均无统计学意义(P>0.05)。结论 孕中期胎儿染色体异常的母体PAPP-A、F-β-HCG水平高,且NIPT联合PAPP-A、F-β-HCG及超声诊断染色体异常有较好的价值。 展开更多
关键词 无创产前DNA检测 血清妊娠相关血浆蛋白-A 游离β-人绒毛膜促性腺激素 染色体异常
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GATA3介导miR-21/PTEN轴对子宫内膜癌细胞增殖、侵袭的影响
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作者 王发辉 邓青春 +1 位作者 林佳佳 陈春妃 《实用医学杂志》 CAS 北大核心 2024年第15期2069-2074,共6页
目的分析GATA结合蛋白3(GATA3)介导微小RNA-21(miR-21)/人类第10号染色体缺失的磷酸酶及张力蛋白同源物(PTEN)轴对子宫内膜癌细胞增殖、侵袭的影响。方法取HEC-1-A细胞,进行转染分组,分为对照组、GATA3空载质粒组、GATA3过表达质粒组、G... 目的分析GATA结合蛋白3(GATA3)介导微小RNA-21(miR-21)/人类第10号染色体缺失的磷酸酶及张力蛋白同源物(PTEN)轴对子宫内膜癌细胞增殖、侵袭的影响。方法取HEC-1-A细胞,进行转染分组,分为对照组、GATA3空载质粒组、GATA3过表达质粒组、GATA3 siRNA阴性对照组、GATA3 siRNA组。检测各组细胞中GATA3、miR-21、PTEN表达量、增殖情况、凋亡率、迁移、侵袭。结果与hEEC组相比,HEC-1-A组、HEC-1-B组、Ishikawa组细胞中GATA3、miR-21表达水平升高,PTEN表达水平降低(P<0.05)。与GATA3空载质粒组相比,GATA3过表达质粒组GATA3、miR-21 mRNA表达量、增殖率、迁移距离、侵袭细胞数、Vimentin水平升高,PTEN mRNA表达量、凋亡率、Caspase-9、Bax、E-cadherin水平降低(P<0.05);与GATA3 siRNA阴性对照组相比,GATA3、miR-21 mRNA表达量、增殖率、迁移距离、侵袭细胞数、Vimentin水平降低,PTEN mRNA表达量、凋亡率、Caspase-9、Bax、E-cadherin水平升高(P<0.05)。结论下调GATA3表达,可对miR-21/PTEN轴进行调节,使HEC-1-A细胞的增殖减慢,促进HEC-1-A细胞的凋亡。 展开更多
关键词 子宫内膜癌 GATA结合蛋白3 微小RNA-21 人类第10号染色体缺失的磷酸酶及张力蛋白同源物 增殖 侵袭
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东方蜜蜂微孢子虫RRDRP的分子特性及系统进化分析
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作者 张凯遥 冯佩林 +6 位作者 宓诗雨 郭思佳 张艺琼 荆欣 陈大福 郭睿 付中民 《福建农林大学学报(自然科学版)》 CAS CSCD 北大核心 2024年第3期371-376,共6页
【目的】通过对东方蜜蜂微孢子虫和其他物种的Rad18-like重组与DNA修复蛋白(Rad18-like recombination and DNA repair protein,RRDRP)进行系统进化分析,为进一步研究其功能提供理论依据。【方法】利用Protparam、ProtScale和SWISS-mode... 【目的】通过对东方蜜蜂微孢子虫和其他物种的Rad18-like重组与DNA修复蛋白(Rad18-like recombination and DNA repair protein,RRDRP)进行系统进化分析,为进一步研究其功能提供理论依据。【方法】利用Protparam、ProtScale和SWISS-model等软件分析RRDRP的理化性质、亲水性和三级结构。利用SignalP 4.1 Server、NetPhos 3.1 Server、TMHMM和SOPMA等软件预测RRDRP的信号肽、磷酸化位点、跨膜结构域及二级结构。使用PSORTⅡ软件预测RRDRP蛋白的亚细胞定位。使用MEME软件预测东方蜜蜂微孢子虫和其他物种RRDRP的保守基序。通过Mega 11.0软件对东方蜜蜂微孢子虫和其他物种的RRDRP进行氨基酸序列多重比对,并采用邻接法构建系统进化树。【结果】东方蜜蜂微孢子虫RRDRP基因含有2928个核苷酸,可编码975个氨基酸,分子质量约11.50 ku,脂溶系数93.58,等电点6.63,分子式C_(5135)H_(8253)N_(1377)O_(1545)S_(34),平均亲水系数为-0.627,可同时定位于细胞核、细胞质、过氧化物酶体和细胞骨架;RRDRP包含55个磷酸化位点、678个α-螺旋、99条延长链、22个β-转角及176个无规则卷曲;RRDRP不含信号肽与跨膜结构域。东方蜜蜂微孢子虫、兔脑炎微孢子虫、颗粒病微粒子虫等11个物种的RRDRP中均含有5个相同的保守基序(Motif 1、Motif 2、Motif 3、Motif 4和Motif 5)。东方蜜蜂微孢子虫与颗粒病微粒子虫在进化树上聚为一支,同源性较高。【结论】东方蜜蜂微孢子虫RRDRP是潜在的亲水性蛋白和胞内蛋白,在东方蜜蜂微孢子虫以及其他微孢子虫和真菌中高度保守。【目的】通过对东方蜜蜂微孢子虫和其他物种的Rad18-like重组与DNA修复蛋白(Rad18-like recombination and DNA repair protein, RRDRP)进行系统进化分析,为进一步研究其功能提供理论依据。【方法】利用Protparam、ProtScale和SWISS-model等软件分析RRDRP的理化性质、亲水性和三级结构。利用SignalP 4.1 Server、NetPhos 3.1 Server、TMHMM和SOPMA等软件预测RRDRP的信号肽、磷酸化位点、跨膜结构域及二级结构。使用PSORT Ⅱ软件预测RRDRP蛋白的亚细胞定位。使用MEME软件预测东方蜜蜂微孢子虫和其他物种RRDRP的保守基序。通过Mega 11.0软件对东方蜜蜂微孢子虫和其他物种的RRDRP进行氨基酸序列多重比对,并采用邻接法构建系统进化树。【结果】东方蜜蜂微孢子虫RRDRP基因含有2 928个核苷酸,可编码975个氨基酸,分子质量约11.50 ku,脂溶系数93.58,等电点6.63,分子式C5135H8253N1377O1545S34,平均亲水系数为-0.627,可同时定位于细胞核、细胞质、过氧化物酶体和细胞骨架;RRDRP包含55个磷酸化位点、678个α-螺旋、99条延长链、22个β-转角及176个无规则卷曲;RRDRP不含信号肽与跨膜结构域。东方蜜蜂微孢子虫、兔脑炎微孢子虫、颗粒病微粒子虫等11个物种的RRDRP中均含有5个相同的保守基序(Motif 1、Motif 2、Motif 3、Motif 4和Motif 5)。东方蜜蜂微孢子虫与颗粒病微粒子虫在进化树上聚为一支,同源性较高。【结论】东方蜜蜂微孢子虫RRDRP是潜在的亲水性蛋白和胞内蛋白,在东方蜜蜂微孢子虫以及其他微孢子虫和真菌中高度保守。 展开更多
关键词 东方蜜蜂微孢子虫 染色体结构维持蛋白 Rad18-like重组和DNA修复蛋白 分子特性 系统进化
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Mechanistic and functional extrapolation of SET and MYND domaincontaining protein 2 to pancreatic cancer 被引量:1
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作者 Eid Alshammari Ying-Xue Zhang Zhe Yang 《World Journal of Gastroenterology》 SCIE CAS 2022年第29期3753-3766,共14页
Pancreatic ductal adenocarcinoma(PDAC)is one of the most lethal neoplasms worldwide and represents the vast majority of pancreatic cancer cases.Understanding the molecular pathogenesis and the underlying mechanisms in... Pancreatic ductal adenocarcinoma(PDAC)is one of the most lethal neoplasms worldwide and represents the vast majority of pancreatic cancer cases.Understanding the molecular pathogenesis and the underlying mechanisms involved in the initiation,maintenance,and progression of PDAC is an urgent need,which may lead to the development of novel therapeutic strategies against this deadly cancer.Here,we review the role of SET and MYND domaincontaining protein 2(SMYD2)in initiating and maintaining PDAC development through methylating multiple tumor suppressors and oncogenic proteins.Given the broad substrate specificity of SMYD2 and its involvement in diverse oncogenic signaling pathways in many other cancers,the mechanistic extrapolation of SMYD2 from these cancers to PDAC may allow for developing new hypotheses about the mechanisms driving PDAC tumor growth and metastasis,supporting a proposition that targeting SMYD2 could be a powerful strategy for the prevention and treatment of PDAC. 展开更多
关键词 Pancreatic ductal adenocarcinoma protein lysine methyltransferase Histone/non-histone methylation Oncogenic signaling pathways Methyltransferase inhibitors
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巴戟天多糖调控精索静脉曲张大鼠睾丸修复的作用机制研究
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作者 余慧 王颖新 +1 位作者 聂丙飞 张健 《世界中医药》 CAS 北大核心 2024年第18期2771-2777,共7页
目的:探讨巴戟天多糖(MOP)对精索静脉曲张型(VC)大鼠睾丸的保护作用。方法:使用左肾静脉缩窄法制备VC模型,将60只大鼠按简单随机法分为空白对照组、模型组、VC组、VC+100 mg/kg巴戟天多糖组、VC+200 mg/kg巴戟天多糖组、VC+300 mg/kg巴... 目的:探讨巴戟天多糖(MOP)对精索静脉曲张型(VC)大鼠睾丸的保护作用。方法:使用左肾静脉缩窄法制备VC模型,将60只大鼠按简单随机法分为空白对照组、模型组、VC组、VC+100 mg/kg巴戟天多糖组、VC+200 mg/kg巴戟天多糖组、VC+300 mg/kg巴戟天多糖组。通过GeneCards数据库筛选VC睾丸修复相关基因;酶联免疫吸附试验法检测促性腺激素释放激素(GnRH)、睾酮(T)、黄体生成素(LH)、卵泡刺激素(FSH)水平;检测左侧睾丸相关指标及活性氧(ROS)含量;TUNEL检测细胞凋亡;蛋白质免疫印迹检测磷酸酯酶与人第10号染色体缺失的磷酸酶及张力蛋白同源的基因蛋白(PTEN)、磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)蛋白表达。结果:相较于模型组,VC组睾丸间质面积、细胞凋亡率、ROS、GnRH、LH、FSH及PI3K、AKT、p-AKT蛋白表达显著升高,睾丸与附睾系数、曲精小管直径、附睾精子数、T及PTEN蛋白表达显著降低(P<0.05);相较于VC组,VC+100 mg/kg组睾丸间质面积、细胞凋亡率、ROS、FSH及PI3K、AKT、p-AKT蛋白表达显著降低,睾丸与附睾系数、曲精小管直径、附睾精子数、T及PTEN蛋白表达显著升高(P<0.05);相较于VC组,VC+200 mg/kg组、VC+300 mg/kg组上述指标结果均相反(P<0.05)。结论:MOP可能通过调控PTEN/PI3K/AKT通路来修复VC大鼠睾丸。 展开更多
关键词 巴戟天多糖 精索静脉曲张 睾丸 修复 生殖细胞 磷酸酯酶 人第10号染色体缺失的磷酸酶及张力蛋白同源的基因蛋白 磷脂酰肌醇3-激酶 蛋白激酶B
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X-染色体连锁凋亡抑制蛋白相关因子1通过自噬途径调控Caspase 1介导的胃肠道间质瘤细胞焦亡
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作者 周江浩 谢书海 陈勇 《河北医学》 CAS 2024年第5期744-750,共7页
目的:探究X-染色体连锁凋亡抑制蛋白相关因子1(XAF1)对人胃肠道间质瘤(GIST)细胞中半胱氨酸蛋白酶-1(Caspase 1)介导的细胞焦亡的影响及机制。方法:实时荧光定量PCR和Western blot检测人正常胃黏膜上皮细胞系(RGM-1)和GIST细胞系(GIST-T... 目的:探究X-染色体连锁凋亡抑制蛋白相关因子1(XAF1)对人胃肠道间质瘤(GIST)细胞中半胱氨酸蛋白酶-1(Caspase 1)介导的细胞焦亡的影响及机制。方法:实时荧光定量PCR和Western blot检测人正常胃黏膜上皮细胞系(RGM-1)和GIST细胞系(GIST-T1、GIST-430、GIST-882)中XAF1的表达水平;采用脂质体介导转染法将pcDNA3.1-XAF1重组质粒染进GIST-882细胞,以构建高表达XAF1的GIST-882细胞。GIST-882细胞分为对照组、pcDNA3.1组(转染pcDNA3.1空载体质粒)、pcDNA3.1-XAF1组(转染pcDNA3.1-XAF1重组质粒)、pcDNA3.1-XAF1+Rap组(转染pcDNA3.1-XAF1重组质粒并给予自噬激活剂雷帕霉素处理),免疫荧光染色检测各组细胞内微管相关蛋白1轻链3(LC3)表达情况,Western blot检测各组细胞中LC3Ⅱ/LC3Ⅰ比值与Beclin1蛋白表达水平,ELISA法检测各组细胞上清中白细胞介素,IL-1β、IL-18含量,乳酸脱氢酶(LDH)释放实验检测各组细胞LDH释放率,Western blot检测各组细胞中Caspase-1及其活化形式cleaved-Caspase-1、NOD样受体热蛋白结构域相关蛋白3(NLRP3)、消皮素D(GSDMD)蛋白表达水平。结果:与RGM-1细胞比较,GIST-T1、GIST-430、GIST-882细胞中XAF1 mRNA和蛋白相对表达量显著减少(P<0.05);细胞转染后,pcDNA3.1-XAF1组GIST-882细胞中XAF1 mRNA和蛋白相对表达量均显著高于pcDNA3.1组、对照组(P<0.05)。与对照组比较,pcDNA3.1-XAF1组GIST-882细胞内LC3荧光强度明显减弱,LC3Ⅱ/LC3Ⅰ比值、Beclin1蛋白相对表达量显著减少(P<0.05),细胞上清液IL-1β、IL-18含量及LDH释放率显著升高(P<0.05),Caspase-1、cleaved-Caspase-1、NLRP3、GSDMD蛋白相对表达量也显著增加(P<0.05);与pcDNA3.1-XAF1组比较,pcDNA3.1-XAF1+Rap组GIST-882细胞内LC3荧光强度明显增强,LC3Ⅱ/LC3Ⅰ比值、Beclin1蛋白相对表达量显著增加(P<0.05),同时,细胞上清液IL-1β、IL-18含量及LDH释放率显著下降(P<0.05),Caspase-1、cleaved-Caspase-1、NLRP3、GSDMD蛋白相对表达量也显著减少(P<0.05)。结论:XAF1在GIST细胞中表达下降,提高XAF1表达能通过抑制自噬从而促进Caspase 1介导的GIST细胞焦亡,发挥抗肿瘤作用。 展开更多
关键词 胃肠道间质瘤 X-染色体连锁凋亡抑制蛋白相关因子1 自噬 半胱氨酸蛋白酶-1 焦亡
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尼美舒利对宫颈癌荷瘤裸鼠移植瘤的影响及机制实验研究
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作者 季滢 郑艳莉 +2 位作者 韩云 孙蓉蓉 朱奕 《陕西医学杂志》 CAS 2024年第8期1026-1030,共5页
目的:探讨尼美舒利对宫颈癌荷瘤裸鼠移植瘤的影响及可能的机制。方法:建立人宫颈癌C33A细胞裸鼠皮下移植瘤模型40只,随机分为模型组、尼美舒利组、过氧化物酶体增殖物激活受体γ(PPARγ)抑制剂组、联合组,每组10只。尼美舒利组每天给予2... 目的:探讨尼美舒利对宫颈癌荷瘤裸鼠移植瘤的影响及可能的机制。方法:建立人宫颈癌C33A细胞裸鼠皮下移植瘤模型40只,随机分为模型组、尼美舒利组、过氧化物酶体增殖物激活受体γ(PPARγ)抑制剂组、联合组,每组10只。尼美舒利组每天给予20 mg/kg尼美舒利灌胃,模型组给予等量0.5%羟甲基纤维素钠灌胃,PPARγ抑制剂组每天给予10 mg/kg PPARγ抑制剂GW9662灌胃,联合组给予尼美舒利联合PPARγ抑制剂处理,均连续干预4周。采用流式细胞术检测各组裸鼠脾脏自然杀伤(NK)细胞活性。采用TUNEL法检测各组裸鼠移植瘤细胞凋亡情况。采用Western blot检测移植瘤组织PPARγ、第10号染色体缺失的磷酸酶及张力蛋白同源的基因(PTEN)、蛋白激酶B(AKT)、p-AKT蛋白表达。结果:40只裸鼠皮下出现直径至少5 mm的肿瘤结节,且未出现红肿或坏死的迹象,模型建立成功。与模型组比较,尼美舒利组NK细胞活性增强,PPARγ抑制剂组NK细胞活性降低(均P<0.05)。与尼美舒利组比较,联合组NK细胞活性降低(P<0.05)。与PPARγ抑制剂组比较,联合组NK细胞活性增强(P<0.05)。各组干预后第3、6、9、12天,与模型组比较,尼美舒利组移植瘤体积缩小,PPARγ抑制剂组移植瘤体积增大(均P<0.05);与尼美舒利组比较,联合组移植瘤体积增大(P<0.05);与PPARγ抑制剂组比较,联合组移植瘤体积缩小(P<0.05)。与模型组比较,尼美舒利组移植瘤细胞凋亡率及PPARγ、PTEN蛋白表达量升高,p-AKT/AKT比值降低;PPARγ抑制剂组移植瘤细胞凋亡率及PPARγ、PTEN蛋白表达量降低,p-AKT/AKT比值升高(均P<0.05)。与尼美舒利组比较,联合组移植瘤细胞凋亡率及PPARγ、PTEN蛋白表达量降低,p-AKT/AKT比值升高(均P<0.05)。与PPARγ抑制剂组比较,联合组移植瘤细胞凋亡率及PPARγ蛋白、PTEN蛋白表达量升高,p-AKT/AKT比值降低(均P<0.05)。结论:尼美舒利可抑制宫颈癌荷瘤裸鼠移植瘤生长,增强NK细胞活性,促进肿瘤细胞凋亡,其作用机制可能与上调PPARγ及PTEN/AKT通路有关。 展开更多
关键词 宫颈癌 移植瘤 尼美舒利 过氧化物酶体增殖物激活受体Γ 第10号染色体缺失的磷酸酶及张力蛋白同源的基因 蛋白激酶B 裸鼠
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乳腺癌组织中PTEN、MMP-2、AKT的表达及其与临床病理特征的相关性
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作者 邓亚萍 荆海红 吴瑜 《实用癌症杂志》 2024年第10期1611-1613,共3页
目的探讨乳腺癌组织内第10号染色体缺失的磷酸酶和张力蛋白同源基因(PTEN)、基质金属蛋白酶-2(MMP-2)、蛋白激酶B(AKT)的表达及其与患者临床病理特征间的关系。方法选取63例乳腺癌患者,收集其癌组织与癌旁正常组织(距离病灶边缘≥3 cm)... 目的探讨乳腺癌组织内第10号染色体缺失的磷酸酶和张力蛋白同源基因(PTEN)、基质金属蛋白酶-2(MMP-2)、蛋白激酶B(AKT)的表达及其与患者临床病理特征间的关系。方法选取63例乳腺癌患者,收集其癌组织与癌旁正常组织(距离病灶边缘≥3 cm),以免疫组织化学法测定组织内PTEN、MMP-2、AKT表达。收集患者的年龄、性别等资料,分析PTEN、MMP-2、AKT表达与患者临床病理特征间的关系。结果癌组织中的MMP-2、AKT阳性表达率高于癌旁组织,PTEN阳性表达率低于癌旁组织,有统计学差异(P<0.05)。PTEN、MMP-2、AKT表达与乳腺癌患者的年龄、肿瘤部位无关(P>0.05);与患者肿瘤的组织学分级、淋巴结转移有关(P<0.05)。结论PTEN、MMP-2、AKT在乳腺癌组织内呈异常表达,其表达与肿瘤的组织学分级、淋巴结转移有关。 展开更多
关键词 乳腺癌 第10号染色体缺失的磷酸酶和张力蛋白同源基因 基质金属蛋白酶-2 蛋白激酶B 临床病理特征
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子宫肌瘤组织中miR-203a-3p及CBX8表达水平及其临床意义
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作者 陈小兰 雷霞 皮发利 《检验医学与临床》 CAS 2024年第20期2958-2964,共7页
目的探究子宫肌瘤组织中微小RNA(miR)-203a-3p及染色体结合蛋白8(CBX8)表达水平及其临床意义。方法选取2019年6月至2021年6月该院收治的112例子宫肌瘤患者作为研究对象,采用实时荧光定量聚合酶链反应检测子宫肌瘤组织与肌瘤旁组织miR-20... 目的探究子宫肌瘤组织中微小RNA(miR)-203a-3p及染色体结合蛋白8(CBX8)表达水平及其临床意义。方法选取2019年6月至2021年6月该院收治的112例子宫肌瘤患者作为研究对象,采用实时荧光定量聚合酶链反应检测子宫肌瘤组织与肌瘤旁组织miR-203a-3p、CBX8 mRNA表达水平,免疫组织化学法检测CBX8蛋白表达。以子宫肌瘤组织中miR-203a-3p表达的中位数为界,将子宫肌瘤患者分为miR-203a-3p低表达组和miR-203a-3p高表达组。以子宫肌瘤组织中CBX8蛋白免疫组化染色的评分为界,将子宫肌瘤患者分为CBX8蛋白阴性表达组和阳性表达组。在Targetscan数据库中进行miR-203a-3p、CBX8靶向关系查询。对112例子宫肌瘤患者术后进行2年随访,根据随访结果将患者分为复发组与未复发组。采用Pearson相关分析子宫肌瘤组织miR-203a-3p表达水平与CBX8 mRNA表达水平的相关性。采用多因素Logistic回归分析子宫肌瘤患者术后复发的影响因素。结果子宫肌瘤组织中miR-203a-3p表达水平明显低于肌瘤旁组织,CBX8 mRNA表达水平明显高于肌瘤旁组织,差异均有统计学意义(P<0.05)。CBX8蛋白阳性率(45.54%,51/112)明显高于肌瘤旁组织(21.43%,24/112),差异有统计学意义(χ^(2)=14.613,P<0.05)。Pearson相关分析结果显示,子宫肌瘤组织中miR-203a-3p表达水平与CBX8 mRNA表达水平呈负相关(r=-0.562,P<0.05)。Targetscan数据库检索结果显示,miR-203a-3p与CBX8存在特异性结合位点。miR-203a-3p低表达组、高表达组均纳入56例患者。miR-203a-3p低表达组与高表达组肿瘤最大径、肌瘤数目、生殖道炎、月经紊乱情况比较,差异均有统计学意义(P<0.05)。CBX8蛋白阴性表达组纳入61例患者,阳性表达组纳入51例患者。CBX8蛋白阴性表达组、阳性表达组肿瘤最大径、肌瘤数目、生殖道炎、月经紊乱情况比较,差异均有统计学意义(P<0.05)。子宫肌瘤患者随访率为100%,术后复发组纳入41例患者,未复发组纳入71例患者,复发率为36.61%。两组肌瘤最大径、肌瘤数目、生殖道炎、月经紊乱情况及CBX8 mRNA、miR-203a-3p水平比较,差异均有统计学意义(P<0.05)。多因素Logistic回归分析结果显示,肌瘤最大径≥3 cm、肌瘤数目多发、CBX8 mRNA表达水平升高、有生殖道炎、有月经紊乱为子宫肌瘤患者术后复发的危险因素(P<0.05),miR-203a-3p表达水平升高为子宫肌瘤患者术后复发的保护因素(P<0.05)。结论子宫肌瘤组织中miR-203a-3p表达水平降低,CBX8 mRNA表达水平升高,与临床病理特征及复发密切相关,可能作为子宫肌瘤诊断以及预后评估的标志物。 展开更多
关键词 子宫肌瘤 微小RNA-203a-3p 染色体结合蛋白8 预后 相关性
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癌-睾丸抗原SPANXB在肝癌中的表达及其影响肝癌进展的机制研究
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作者 薛煜 张海龙 雷鸣 《上海交通大学学报(医学版)》 CAS CSCD 北大核心 2024年第7期801-813,共13页
目的·分析癌-睾丸抗原(cancer-testis antigen,CTA)家族成员SPANXB(sperm protein associated with the nucleus on the X chromosome B)在肝癌中的表达及其与肝癌患者预后之间的相关性,并探究SPANXB对肝癌细胞增殖的影响及其潜在... 目的·分析癌-睾丸抗原(cancer-testis antigen,CTA)家族成员SPANXB(sperm protein associated with the nucleus on the X chromosome B)在肝癌中的表达及其与肝癌患者预后之间的相关性,并探究SPANXB对肝癌细胞增殖的影响及其潜在机制。方法·利用癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中的肝癌样本数据,分析SPANXB在肝癌组织中的表达及其与患者生存期的相关性。构建稳定敲低SPANXB与稳定过表达SPANXB的肝癌细胞系,利用活细胞成像实验、EdU细胞增殖实验和平板克隆形成实验评估SPANXB对肝癌细胞增殖的影响。通过RNA测序(RNA-sequence,RNA-seq)探究SPANXB调控肝癌细胞增殖的相关通路,并利用细胞周期实验验证SPANXB对肝癌细胞周期的影响。采用免疫沉淀-质谱联用技术(immunoprecipitation-mass spectrometry,IP-MS)探索与SPANXB相互作用的蛋白,并使用免疫共沉淀(co-immunoprecipitation,Co-IP)进行验证。结果·SPANXB mRNA在肝癌组织中的表达高于正常组织(P=0.003),且与肝癌患者的生存期呈负相关。稳定敲低SPANXB可降低肝癌细胞的增殖能力、克隆形成能力,而稳定过表达SPANXB则可促进这些过程。RNA-seq的结果显示,SPANXB的敲低可下调DNA复制与G1/S细胞周期转换相关通路,细胞周期实验的结果显示SPANXB的敲低可导致肝癌细胞周期发生改变。IP-MS和Co-IP结果显示,SPAXNB与有丝分裂停滞缺陷2样蛋白1(mitotic arrest deficient 2-like protein 1,MAD2L1)、WD重复域蛋白5(WD repeat domain 5,WDR5)等细胞周期相关蛋白存在相互作用。结论·SPANXB的高表达与肝癌的预后呈负相关,其可能通过与MAD2L1、WDR5相互作用调控细胞周期并增强肝癌细胞的增殖活性。 展开更多
关键词 癌-睾丸抗原 SPANXB 肝癌 细胞周期
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非染色体结构维护凝集素Ⅰ复合物亚基G通过Akt信号通路促进卵巢癌细胞的增殖、迁移和侵袭
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作者 张明姝 王艺慧 +1 位作者 张晴 叶丽平 《解剖学报》 CAS CSCD 2024年第5期573-581,共9页
目的探讨非染色体结构维护凝集素Ⅰ复合物亚基G(NCAPG)调控卵巢癌细胞的增殖、迁移和侵袭的机制。方法生物信息学GEPIA数据库分析NCAPG在卵巢癌组织中的差异表达。Western blotting检测正常卵巢上皮细胞IOSE80、卵巢癌A2780和SKOV3细胞... 目的探讨非染色体结构维护凝集素Ⅰ复合物亚基G(NCAPG)调控卵巢癌细胞的增殖、迁移和侵袭的机制。方法生物信息学GEPIA数据库分析NCAPG在卵巢癌组织中的差异表达。Western blotting检测正常卵巢上皮细胞IOSE80、卵巢癌A2780和SKOV3细胞中NCAPG的蛋白表达。NCAPG siRNA沉默实验分为空白组、对照组、siNCAPG-1组和siNCAPG-2组。NCAPG过表达实验分为空白组、对照组、NCAPG组、NCAPG+MK2206组和MK2206组。MTT实验检测细胞增殖活性;细胞划痕实验和Transwell实验评估细胞的迁移和侵袭能力;Western blotting检测细胞的磷酸化Akt(p-Akt)、总Akt(t-Akt)、增殖细胞核抗原(PCNA)、基质金属蛋白酶9(MMP-9)、波形蛋白(vimentin)、N-钙黏蛋白(N-cadherin)和E-钙黏蛋白(E-cadherin)的表达水平。结果NCAPG在卵巢癌组织和卵巢癌细胞中高表达。沉默NCAPG可明显抑制卵巢癌SKOV3细胞的增殖、迁移和侵袭,p-Akt、PCNA、MMP-9、vimentin和N-cadherin表达减少,E-cadherin表达增多。过表达NCAPG质粒可促进卵巢癌A2780细胞增殖、迁移和侵袭,p-Akt、PCNA、MMP-9、vimentin和N-cadherin表达增多,E-cadherin表达降低。Akt抑制剂MK2206可明显抑制NCAPG的上述作用。结论NCAPG激活Akt信号通路,调控PCNA、MMP-9和上皮细胞-间充质转化(EMT)相关蛋白的表达,促进卵巢癌细胞的增殖、迁移和侵袭。 展开更多
关键词 卵巢癌 非染色体结构维护凝集素Ⅰ复合物亚基G 蛋白激酶B信号通路 增殖细胞核抗原 基质金属蛋白酶9 钙黏蛋白 免疫印迹法
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依托咪酯通过下调含WW结构域的E3泛素蛋白连接酶2表达抑制非小细胞肺癌A549细胞增殖并诱导其凋亡的实验研究
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作者 何元 段晓飞 +1 位作者 党莎杰 王培 《中国医药》 2024年第11期1640-1644,共5页
目的探讨依托咪酯(ETO)对非小细胞肺癌(NSCLC)A549细胞增殖和凋亡的影响及含WW结构域的E3泛素蛋白连接酶2(WWP2)在其中发挥的作用机制。方法分别采用0、1、2和3 mg/L的ETO处理人正常肺上皮细胞系BESA-2B和人NSCLC细胞系A549,细胞计数试... 目的探讨依托咪酯(ETO)对非小细胞肺癌(NSCLC)A549细胞增殖和凋亡的影响及含WW结构域的E3泛素蛋白连接酶2(WWP2)在其中发挥的作用机制。方法分别采用0、1、2和3 mg/L的ETO处理人正常肺上皮细胞系BESA-2B和人NSCLC细胞系A549,细胞计数试剂盒法检测细胞活力;将A549细胞随机分为对照组、ETO(3 mg/L)组、ETO+NC组和ETO+WWP2-OE组,后2组分别于ETO处理后转染空载体pcDNA3.1-NC或WWP2过表达载体pcDNA3.1-WWP2,集落形成试验检测细胞增殖能力;TUNEL染色检测细胞凋亡;实时荧光定量聚合酶链反应法检测各组细胞中WWP2的mRNA表达;STITCH数据库选择与ETO直接相互作用的蛋白质;蛋白质印迹法检测各组细胞中WWP2、增殖、凋亡和磷酸酶与张力蛋白同源物(PTEN)/磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(Akt)通路相关蛋白表达。结果ETO以剂量依赖性方式显著降低A549细胞活力(F=147.923,P<0.001);而对正常肺上皮BESA-2B细胞活力无影响(F=1.427,P=0.126)。不同浓度(0、1、2、3 mg/L)ETO处理A549细胞后,集落形成数量逐渐减少[0、1、2、3 mg/L ETO组分别为(898±38)、(785±48)、(635±36)、(388±20)个],细胞凋亡率逐渐升高[0、1、2、3 mg/L ETO组分别为(2.23±0.65)%、(7.63±0.35)%、(13.24±0.47)%、(18.93±0.36)%],呈剂量依赖性(F=218.732、352.786,均P<0.001)。STITCH数据库预测ETO可通过上调WWP2蛋白表达和下调PTEN蛋白表达与WWP2和PTEN相互作用。与对照组相比,ETO组细胞中WWP2的mRNA和蛋白表达降低,集落形成数量减少,细胞凋亡率升高,增殖细胞核抗原(PCNA)、细胞增殖抗原Ki67、B淋巴细胞瘤基因2(Bcl-2)和PTEN蛋白表达降低,Bcl-2相关X蛋白(Bax)和含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved caspase-3)蛋白表达升高,磷酸化PI3K(p-PI3K)/PI3K和磷酸化Akt(p-Akt)/Akt比值降低(均P<0.01);与ETO+NC组比较,ETO+WWP2-OE组细胞中WWP2的mRNA和蛋白表达升高,集落形成数量增多,细胞凋亡率降低,PCNA、Ki67、Bcl-2和PTEN蛋白表达增多,Bax和Cleaved caspase-3蛋白表达减少,p-PI3K/PI3K和p-Akt/Akt比值升高(均P<0.01)。结论ETO可抑制A549细胞增殖并促进细胞凋亡,其作用机制可能与调控WWP2的下调和PTEN/PI3K/Akt通路的激活有关。 展开更多
关键词 非小细胞肺癌 依托咪酯 含WW结构域的E3泛素蛋白连接酶2 磷酸酶与张力蛋白同源物/磷脂酰肌醇-3-激酶/蛋白激酶B通路 增殖 凋亡
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Copy number changes of target genes in chromosome 3q25.3-qter of esophageal squamous cell carcinoma: TP63is amplified in early carcinogenesis but down-regulated as disease progressed 被引量:5
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作者 Chueh-ChuanYen Yann-JangChen +12 位作者 Chin-ChenPan Kai-HsiLu PaulChih-HsuehChen Jiun-YiHsia Jung-TaChen Yu-ChungWu Wen-HuHsu Liang-ShunWang Min-HsiungHuang Biing-ShiungHuang Cheng-PoHu Po-MinChen Chi-HungLin 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第9期1267-1272,共6页
AIM: By using comparative genomic hybridization, gain of 3q was found in 45-86% cases of esophageal squamous cell carcinoma (EC-SCC). Chromosome 3q25.3-qter is the minimal common region with several oncogenes found wi... AIM: By using comparative genomic hybridization, gain of 3q was found in 45-86% cases of esophageal squamous cell carcinoma (EC-SCC). Chromosome 3q25.3-qter is the minimal common region with several oncogenes found within this region. However, amplification patterns of these genes in EC-SCC have never been reported. The possible association of copy number changes of these genes with pathologic characteristics is still not clear. METHODS: Real-time quantitative PCR (Q-PCR) was performed to analyze the copy number changes of 13 candidate genes within this region in 60 primary tumors of EC-SCC, and possible association of copy number changes with pathologic characteristics was analyzed by statistics. Immunohistochemistry (IHC) study was also performed on another set of 111 primary tumors of EC-SCC to verify the association between TP63 expression change and lymph node metastasis status. RESULTS: The average copy numbers (±SE) per haploid genome of individual genes in 60 samples were (from centromere to telomere): SSR3: 4.19 (±0.69); CCNL1: 5.24 (±0.67); SMC4L1: 2.01 (±0.16); EVI1: 2.02 (±0.12); hTERC. 5.28 (±0.54); SKIL 2.71 (±0.14); EIF5A2. 1.95 (±0.12); ECT2: 9.18 (±1.68); PIK3CA: 8.13 (±1.17); EIF4G1: 1.07 (±0.05); 557: 3.07 (±0.25); TP63: 2.51 (±0.22); TFRC. 2.42 (±0.19). Four clusters of amplification were found: SSR3 and CCLN1 at 3q25.31; hTERC and SKIL at 3q26.2; ECT2 and PIK3CA at 3q26.31-q26.32; and 55T, TP63 and TFRC at 3q27.3-q29. Patients with lymph node metastasis had significantly lower copy number of TP63 in the primary tumor than those without lymph node metastasis. IHC study on tissue arrays also showed that patients with lymph node metastasis have significantly lower TP63 staining score in the primary tumor than those without lymph node metastasis. CONCLUSION: This study showed that different amplification patterns were seen among different genes within 3q25.3-qter in EC-SCC, and several novel candidate oncogenes (SSR3, SMC4L1, ECT2, and SST) were identified. TP63 is amplified in early stage of EC-SCC carcinogenesis but down-regulated in advanced stage of disease. 展开更多
关键词 chromosomal aberration Comparative genomic hybridization Esophageal neoplasm IMMUNOHISTOCHEMISTRY Quantitative real-time PCR Tissue array Tumor protein 63
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ISOLATION AND CHARACTERIZATION OF CHROMOSOMAL HMG PROTEINS FROM PEA
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作者 徐典朋 杨中汉 《Chinese Science Bulletin》 SCIE EI CAS 1982年第8期880-883,共4页
An extract from chromatin with 0.35 M NaCl is a heterogenous class of nonhistone chromosomal proteins. On the basis of their respective electrophoretic mobilities in polyacrylamide gel, the 0.35 M NaCl extract is divi... An extract from chromatin with 0.35 M NaCl is a heterogenous class of nonhistone chromosomal proteins. On the basis of their respective electrophoretic mobilities in polyacrylamide gel, the 0.35 M NaCl extract is divided into 2 groups: the low mobility group (LMG) proteins and the high mobility group (HMG) proteins. Up to date, a lot of researches have been mainly concentrated on the latter. It is thought 展开更多
关键词 chromosomal POLYACRYLAMIDE latter respective CHROMATIN wheat proteins residues concentrated AMMONIUM
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The Evolution of Microtubule End-Binding Protein 1 (EB1) and Roles in Regulating Microtubule Behavior
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作者 Jiayu Liu Rong Han 《American Journal of Plant Sciences》 2015年第13期2114-2121,共8页
All organisms must transmit genetic information to offspring through cell division, and mitotic spindle participates in the process. Spindle dynamics through depolymerization or polymerization of microtubules generate... All organisms must transmit genetic information to offspring through cell division, and mitotic spindle participates in the process. Spindle dynamics through depolymerization or polymerization of microtubules generates the driving force required for chromosome movements in mitosis. To date, studies have shown that microtubule arrays control the directions of cell division and diverse microtubule-associated proteins regulate cell division. But a clear picture of how microtubules and microtubule-associated proteins modulate cell division remains unknown. Depletion of end-binding protein 1 by RNA-mediated inhibition shows that one of the microtubule-associated proteins, end-binding protein 1, plays a crucial role in mitotic spindle formation and promotes microtubule dynamics and is needed for the proper segregation of mitotic chromosomes during anaphase in Drosophila cells. Here, we review the properties of end-binding protein 1 and the roles of end-binding protein 1 in regulating microtubule behavior and in cell cycle. 展开更多
关键词 MICROTUBULES End-Binding protein 1 chromosomE SEGREGATION
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