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蛋白激酶Cα对人正常肝和肝癌细胞中Ha-ras基因表达的影响

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生物化学与生物物理进展 2002年第5期29卷 781-785页

作者：冯怡柳惠图高萍北京师范大学生命科学学院细胞增殖及调控生物学教育部重点实验室北京100875

运用基因转染技术 ,将蛋白激酶C(PKCα)cDNA正向插入的真核表达重组质粒pXJ4 1 PKCα ,导入人正常肝细胞 (L 0 2 ) ,经蛋白质免疫印迹等检验 ,表明成功构建了稳定过表达PKCα的人正常肝细胞模型 (LT3) ,用LT3和表达反义PKCα的人肝癌细... 详细信息

运用基因转染技术 ,将蛋白激酶C(PKCα)cDNA正向插入的真核表达重组质粒pXJ4 1 PKCα ,导入人正常肝细胞 (L 0 2 ) ,经蛋白质免疫印迹等检验 ,表明成功构建了稳定过表达PKCα的人正常肝细胞模型 (LT3) ,用LT3和表达反义PKCα的人肝癌细胞HT6为细胞模型 ,通过RT PCR ,蛋白质印迹 (Westernblot)等分析和进一步运用自行构建的真核表达质粒prasGL3,进行荧光素酶活性检测表明 :过表达PKCα可以促进L 0 2细胞的增殖速率 ,并引起Ha ras基因转录水平上升和Ha ras启动子活性升高 ;反之 ,表达反义PKCα的BEL 74 0 2细胞增殖被抑制 ,Ha ras基因转录水平下降 ,Ha ras启动子活性降低 .通过实验我们首次观察到 ,PKCα亚类对Ha ras癌基因表达的影响与其对Ha ras启动子活性的作用有关 ,PKCα可能参与了对Ha

关键词：蛋白激酶Cα 人正常肝肝癌细胞 Ha-ras 基因表达

Effects of overexpressing p14^(ARF) on the apoptosis in human melanoma cells irradiated with γ-ray

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Chinese Science Bulletin 2003年第18期48卷 1971-1975页

作者： PENG Lixia, zhang wei, liu huitu, HE Dacheng & GAO Ping key Laboratry of cell proliferation and regulation biology of ministry of education, college of life Science, beijing normal university, Bei-jing 100875, china Correspondence should be addressed to liu huitu (e-mail: liuhuitu@ ***) key Laboratry of cell proliferation and regulation biology of ministry of education College of Life Science Beijing Normal University Beijing 100875 China

tumor suppressor ARF can induce cell cycle arrest or apoptosis by activating p53. In order to explore the molecular mechanism of the induction of apoptosis by p14ARF, a human melanoma cell model overexpressing p14ARF was constructed. Present study indicated that in the cells overex-pressing p14ARF, p53 was accumulated in nucleus while it dispersed in cytosol in the control cells. Irradiated with g-ray, overexpressing p14ARF promoted the apoptosis of A375 cells, triggered Smac release from mitochondria to cytosol, and increased the expression of p53, Bax, Caspase-3, Caspase-9, p21cip1 and p27kip1. However, the protein level of Bcl-2 and phospho-ERK was down-regulated. These results suggested a possible mechanism of p14ARF in promotion of apoptosis.

关键词： p14ARF p53 g-ray apoptosis A375 cells.

PKC_γ过表达诱导C_3H_(10)T_(1/2)细胞生长失控的初探

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生物化学与生物物理进展 1999年第6期26卷 569-573页

作者：赵霞柳惠图北京师范大学生物系细胞增殖及调控生物学开放实验室北京100875

通过ＤＮＡ重组构建蛋白激酶Ｃγ（ＰＫＣγ）亚类的重组质粒并经基因转染技术和ＤＮＡ印迹、蛋白质印迹与ＰＫＣ活性分析，获得了过表达ＰＫＣγ的Ｃ３Ｈ１０Ｔ１／２细胞———ＮＣＰ４．ＮＣＰ４细胞生长速率提高，流式... 详细信息

通过ＤＮＡ重组构建蛋白激酶Ｃγ（ＰＫＣγ）亚类的重组质粒并经基因转染技术和ＤＮＡ印迹、蛋白质印迹与ＰＫＣ活性分析，获得了过表达ＰＫＣγ的Ｃ３Ｈ１０Ｔ１／２细胞———ＮＣＰ４．ＮＣＰ４细胞生长速率提高，流式细胞光度术检测表明，ＮＣＰ４细胞Ｇ１期百分率下降，Ｓ期和Ｇ２＋Ｍ期百分率升高，与对照组细胞相比，血清依赖性明显下降，贴壁依赖性降低，在软琼脂中形成小集落，出现部分转化表型．进一步检测，首次观察到ＮＣＰ４细胞中癌基因ｃｓｉｓ表达明显增强，这可能是ＮＣＰ４细胞血清依赖性下降的分子机理之一．实验表明，在正常Ｃ３Ｈ１０Ｔ１／２细胞中ＰＫＣγ的过表达可直接导致细胞增殖加速并可诱导出现部分转化特征．

关键词：细胞增殖细胞转化 PKCγ C3H10T1/2 调控

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ARF——一种新的抑癌因子的研究进展

生物化学与生物物理进展 2002年第6期29卷 842-846页

作者：彭丽霞张伟何大澄柳惠图北京师范大学生命科学学院教育部细胞增殖与调控生物学重点实验室北京100875

INK4a/ARF基因位于人染色体 9p2 1,是人类肿瘤中最常见的基因失活位点之一 .INK4a/ARF基因有两套各自独立的启动子 ,通过可变阅读框 ,能够编码两种蛋白质 :p16 INK4a和p14 ARF (ARF在鼠细胞中为p19ARF) .p16作为CDK4 /6的抑制因子 ,能... 详细信息

INK4a/ARF基因位于人染色体 9p2 1,是人类肿瘤中最常见的基因失活位点之一 .INK4a/ARF基因有两套各自独立的启动子 ,通过可变阅读框 ,能够编码两种蛋白质 :p16 INK4a和p14 ARF (ARF在鼠细胞中为p19ARF) .p16作为CDK4 /6的抑制因子 ,能够阻断pRb磷酸化 ,将细胞周期阻断在G1期 ;而ARF可结合原癌蛋白MDM2 ,稳定 p5 3,将细胞周期阻断在G1期和G2 /M转换期 ,或诱导细胞凋亡 .因此ARF蛋白和

关键词： ARF 抑癌因子研究进展细胞周期阻断细胞凋亡

p14^(ARF)对人黑色素瘤细胞增殖的影响及其作用机理的初探

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生物化学与生物物理进展 2004年第2期31卷 177-180页

作者：彭丽霞张伟柳惠图何大澄高萍北京师范大学生命科学院教育部细胞增殖与调控生物学重点实验室北京100875

ARF (alternativereadingframe)作为INK4a/ARF的 β转录产物 ,能够稳定p5 3,诱导细胞周期阻断或凋亡 .利用高表达p1 4 ARF的人黑色素瘤细胞模型 ,探讨了ARF抑制细胞增殖的分子作用机理 .研究发现p1 4 ARF高表达能将细胞周期阻断在G1和G2... 详细信息

ARF (alternativereadingframe)作为INK4a/ARF的 β转录产物 ,能够稳定p5 3,诱导细胞周期阻断或凋亡 .利用高表达p1 4 ARF的人黑色素瘤细胞模型 ,探讨了ARF抑制细胞增殖的分子作用机理 .研究发现p1 4 ARF高表达能将细胞周期阻断在G1和G2期 ,p5 3,p2 1 cip1 和p2 7kip1 蛋白水平明显增强 ,而p ERK1 / 2 ,CyclinD1和CyclinE蛋白水平下降 ,明显抑制细胞生长 .提示p1 4 ARF能通过ERK (extracellularsignal regulatedkinase)

关键词：人黑色素瘤细胞增殖作用机理 p14^ARF 细胞周期

Discovery of laryngeal carcinoma by serum proteomic pattern analysis

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Science china(life sciences) 2004年第3期47卷 219-223页

作者： XIAO Xueyuan1, ZHAO Xiaodong2, liu Jiankai3, GUO Fuzheng1, liu Danhui1 & HE Dacheng1 1. key laboratory for cell proliferation and regulation biology, ministry of education, beijing normal university, Universities?Confeder-ated Institute of Proteomics, beijing 100875, china 2. Department of Otolaryngology-Head and Neck Surgery, Third Affiliated Hospital, Jilin university, Changchun 130031, china 3. Department of Biochemistry, Jilin university, Changchun 130021, china 1. key laboratory for cell proliferation and regulation biology Ministry of Education Beijing Normal University Universities’ Confed ated Institute of Proteomics 100875 Beijing China 2. Department of Otolaryngology-Head and Neck Surgery Third Affiliated Hospital Jilin University 130031 Changchun China 3. Department of Biochemistry Jilin University 130021 Changchun China

Laryngeal carcinoma is the most common malignancy among head and neck tu-mors. The purpose of this study is to find biomarkers for laryngeal carcinoma in patient blood serum using the Surface Enhanced Laser Desorption/Ionization (SELDI) technique. Serum samples from 33 laryngeal carcinoma (12 cases of glottis, 18 of supraglottis and 3 of subglottis) patients and 31 age- and sex-matched healthy people were analyzed by SELDI-TOF on a Pro-teinChip reader, PBSII-C. Protein profiles were generated using WCX2 protein chips. Protein peak clustering and classification analyses were performed utilizing the Biomarker Wizard and Biomarker Pattern software packages, respectively. The results showed that sixteen peaks had significant difference between laryngeal cancer patients and healthy group, eight of which were up-regulated in the patient samples, and the others were down-regulated. Two protein peaks 8153 Da and 2035 Da were automatically chosen for the system training and development of a classification tree. The analysis yielded a correct percentage of 96.9% for patients and 96.7% for control. The results suggest that serum is a useful resource for the detection of specific bio-markers for laryngeal carcinoma. Proteinchip Array System was a useful tool for a high throughput screening of large-sized serum samples to discover potential biomarkers for carci-noma.

关键词： protein chip array system, laryngeal carcinoma, biomarker pattern.

Serum Amyloid A Protein: A Potential Biomarker Correlated With Clinical Stage of Lung Cancer

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Biomedical and Environmental sciences 2007年第1期20卷 33-40页

作者： DAN-hui liu XIAO-MIN WANG LI-JUAN zhang SONG-wei DAI LI-YUN liu JI-FU liu SHAN-SHAN WU SHUAN-YING YANG SAM FU XUE-YUAN XIAO DA-CHENG HE key laboratory of cell proliferation and regulation of ministry of education Beijing Normal University Beijing 100875 China Department of Thoracic Surgery General Hospital of Beijing Command PLA Beijing 100700 China Department of Respiratory Medicine Second Hospital of Xi＇an Jiaotong University Xi＇an 710004 Shaanxi China Ciphergen Biosystems Inc. Fremont CA 94555 USA

Objective To identify serum diagnosis or progression biomarkers in patients with lung cancer using protein chip profiling analysis. Method Profiling analysis was performed on 450 sera collected from 213 patients with lung cancer, 19 with pneumonia, 16 with pulmonary tuberculosis, 65 with laryngeal carcinoma, 55 with laryngopharyngeal carcinoma patients, and 82 normal individuals. A new strategy was developed to identify the biomarkers on chip by trypsin pre-digestion. Results Profiling analysis demonstrated that an 11.6kDa protein was significandy elevated in lung cancer patients, compared with the control groups （P〈0.001）. The level and percentage of 11.6kDa protein progressively increased with the clinical stages Ⅰ-Ⅳ and were also higher in patients with squamous cell carcinoma than in other subtypes. This biomarker could be decreased after operation or chemotherapy. On the other hand, 11.6kDa protein was also increased in 50% benign diseases of lung and 13% of other cancer controls. After trypsin pre-digestion, a set of new peptide biomarkers was noticed to appear only in the samples containing a 11.6kDa peak. Further identification showed that 2177Da was a fragment of serum amyloid A （SAA, MW 11.6kDa）. Two of the new peaks, 1550Da and 1611Da, were defined from the same protein by database searching. This result was further confirmed by partial purification of 11.6kDa protein and MS analysis. Conclusion SAA is a useful biomarker to monitor the progression of lung cancer and can directly identify some biomarkers on chip.

关键词： Lung cancer Serum amyloid A On chip identification Surface enhanced laser desorption/ionization Biomarker

Cooperation-based sperm clusters mediate sperm oviduct entry and fertilization

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Protein & cell 2021年第10期12卷 810-817页

作者： Yongcun Qu Qi Chen Shanshan Guo Chiyuan Ma Yonggang Lu Junchao Shi Shichao liu Tong Zhou Taichi Noda Jingjing Qian Liwen zhang Xili Zhu Xiaohua Lei Yujing Cao wei Li wei Li Nicolas Plachta Martin M.Matzuk Masahito Ikawa Enkui Duan Ying zhang Hongmei Wang State key laboratory of Stem cell and Reproductive biology Institute of ZoologyChinese Academy of SciencesBeijing 100101China university of Chinese Academy of sciences Beijing 100049China Division of Biomedical sciences School of MedicineUniversity of CaliforniaRiversideCA 92521USA Research Institute for Microbial Diseases Osaka UniversitySuitaOsakaJapan Department of Physiology and cell biology Reno School of MedicineUniversity of NevadaRenoNVUSA Institute of Molecular and cell biology A*STARSingaporeSin gapore Center for Drug Discovery Department of Pathology and ImmunologyBaylor College of MedicineHoustonTX 77030USA Division of biology and Biological Engineering California Institute of TechnologyPasadenaCAUSA Institute for Stem cell and Regeneration Chinese Academy of SciencesBeijing 100101China The key laboratory of cell proliferation and regulation biology Ministry of EducationInstitute of Cell BiologyBeijing Normal UniversityBeijing 100875China

Dear Editor,Sperm cooperation has been observed in multiple species(Pizzari and Foster,2008),yet its existenee and benefit for reproductive success in mammals remains ***,combining tissue-clearing with deep three-dimensional(3D)imagi ng,we dem on strate that postcopulatory mouse sperm congregate into unidirectional sperm cooperative clusters at the utero-tubal junction(UTJ),a key physical barrier for passage into the oviduct.

关键词： passage media Foster

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新型Th9细胞分化及功能调控的研究进展

中国免疫学杂志 2019年第5期35卷 513-520页

作者：刘芮辰董琳刘光伟北京师范大学生命科学学院教育部细胞增殖与调控生物学重点实验室北京100875

Th9细胞是近年来新定义的一类辅助性T细胞亚群,主要由转化生长因子β(TGF-β)和白细胞介素4(IL-4共同诱导初始CD4^+T细胞分化而来。本文较详尽综述了Th9细胞分化过程中的分子调控机制及其在抗肿瘤免疫、炎症性肠病和过敏性疾病等热点研... 详细信息

Th9细胞是近年来新定义的一类辅助性T细胞亚群,主要由转化生长因子β(TGF-β)和白细胞介素4(IL-4共同诱导初始CD4^+T细胞分化而来。本文较详尽综述了Th9细胞分化过程中的分子调控机制及其在抗肿瘤免疫、炎症性肠病和过敏性疾病等热点研究中的最新研究进展和重要作用。深入研究Th9细胞分化及功能调控将对阐明多种重要免疫相关疾病的致病机理和防治策略提供依据。

关键词： Th9细胞分化转录因子信号抗肿瘤免疫炎症性肠疾病