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KAI1 inhibits HGF-induced invasion of pancreatic cancer by sphingosine kinase activity

Hepatobiliary & Pancreatic Diseases International 2011年第2期10卷 201-208页

作者： Xu Liu,Xiao-Zhong Guo,Wei-Wei Zhang,Zhuo-Zhuang Lu,Qun-Wei Zhang, Hai-Feng Duan and Li-Sheng Wang State Key Laboratory of Cancer Biology and institute of Digestive Diseases,Xijing Hospital of Digestive Disease,Fourth Military Medical University,Xi’an 710032,china department of Gastroenterology,Shenyang General Hospital of PLA,Shenyang 110016, china department of experimental hematology,beijing institute of radiation medicine,beijing 100850,china [a] State Key Laboratory of Cancer Biology and institute of Digestive Diseases Xijing Hospital of Digestive Disease Fourth Military Medical University Xi'an 710032 China [b] department of Gastroenterology Shenyang General Hospital of PLA Shenyang 110016 China [c] department of experimental hematology Beijing Institute of Radiation Medicine Beijing 100850 China

BACKGROUND:KAI1/CD82 has been reported to attenuate the process of metastases in a variety of tumors;however,its mechanism of action in invasion has not been fully elucidated. The present study aimed to investigate the importance of KAI1 in invasion and its correlation with activation of sphingosine kinase(SPK)in human pancreatic cancer PANC1 and Miapaca-2 cell lines. METHODS:The expression of KAI1 in PANC1 and Miapaca-2 cells,which was mediated by recombinant adenovirus(Ad-KAI1), was assessed by a flow cytometer and Western *** successful infection was established,in vitro growth curve and invasive ability in Boyden Chamber assay were *** presence of KAI1 correlating with c-Met and SPK was detected by co-immunoprecipitation and[γ-32P]ATP incorporation. RESULTS:KAI1 genes had no significant effects on the curve representing cell *** infection with the KAI1 gene,decreased invasive ability in the Boyden Chamber assay was observed in PANC1 and Miapaca-2 cells that were induced by hepatocyte growth ***-expression of KAI1 in the cells led to the deactivation of SPK and a decreased level of intracellular *** correlation was observed between c-Met and KAI1 during co-immunoprecipitation. CONCLUSION:The results of this study for the first time demonstrated a regulatory role for KAI1 in SPK activation, which leads to decreased invasive ability in disease progression of human pancreatic cancer.

关键词： adenovirus vector KAI1 gene sphingosine kinase pancreatic cancer cell lines

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KAI1/CD82 suppresses hepatocyte growth factorinduced migration of hepatoma cells via upregulationof Sprouty2

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Science china(Life Sciences) 2008年第7期51卷 648-654页

作者： MU ZhenBin1 ,3,WANG Hua 2,ZHANG Jing 2,LI QingFang 2,WANG LiSheng 2&GUO XiaoZhong 3 1State Key Laboratory of Cancer Biology and institute of Digestive Diseases,Xijing Hospital,Fourth Military Medical University,Xi’an 710032,china 2department of experimental hematology,beijing institute of radiation medicine,beijing 100850,china 3department of Gastroenterology,Shenyang General Hospital of PLA,Shenyang 110016,china 1. State Key Laboratory of Cancer Biology and institute of Digestive Diseases Xijing Hospital Fourth Military Medical University Xi’an 710032 China 3. department of Gastroenterology Shenyang General Hospital of PLA Shenyang 110016 China 2. department of experimental hematology Beijing Institute of Radiation Medicine Beijing 100850 China

We conducted a study concerning the suppressive mechanism of KAI1/CD82 on hepatoma cell *** growth factor(HGF)induces the migration of hepatoma cells through activation of cellular sphingosine kinase 1(SphK1).Adenovirus-mediated gene transfer of KAI1(Ad-KAI1)downregulates the SphK1 expression and suppresses the HGF-induced migration of SMMC-7721 human hepatocellcular carcinoma *** of KAI1/CD82 significantly elevates Sprouty2 at the protein *** of Sprouty2 with RNA interference can block the KAI1/CD82-induced suppression of hepatoma cell migration and downregulation of SphK1 *** is demonstrated that KAI1/CD82 suppresses HGF-induced migration of hepatoma cells via upregulation of Sprouty2.

关键词： KAI1/CD82 Sprouty2 migration hepatocyte growth factor

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Recruitment of stem cells by hepatocyte growth factor via intracoronary gene transfection in the postinfarction heart failure

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Science china(Life Sciences) 2007年第6期50卷 748-752页

作者： YANG ZhiJian1, WANG Wei1, MA DongChao1, ZHANG YouRong2, WANG LianSheng1, ZHANG YuQing1, XU ShunLin1, CHEN Bo1, MIAO DengShun3, CAO KeJiang1 & MA WenZhu1 1 department of Cardiovascular medicine, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, china 2 department of experimental hematology, beijing institute of radiation medicine, beijing 100850, china 3 department of experimental anatomy, Nanjing Medical University, Nanjing 210029, china department of Cardiovascular medicine the First Affiliated Hospital of Nanjing Medical University Nanjing 210029 China department of experimental hematology Beijing Institute of Radiation Medicine Beijing 100850 China department of experimental anatomy Nanjing Medical University Nanjing 210029 China

We aim to study the amelioration effect of adenovirus5-mediated human hepatocyte growth factor gene transfer on postinfarction heart failure in swine model. Twelve Suzhong young swine were randomly divided into 2 groups of 6 pigs each: Ad5-HGF group and mock-vector Ad5 group. Four weeks after ligation of the left anterior descending coronary artery, Ad5-HGF was intracoronarily transferred into the myocardium. Simultaneously, gate cardiac perfusion imaging was performed to evaluate the heart function. Three weeks later, gate cardiac perfusion imaging was performed again, then the hearts were removed and sectioned for immunohistochemical examination to illustrate the effects of Ad5-HGF on infarcted myocardium. The expression of HGF was examined by ELISA. The results were: (1) compared with the mock-vector Ad5 group, high expression of human HGF was observed in the myocardium of Ad5-HGF group; (2) in the Ad5-HGF group, the number of CD117+ cells co-expressing c-Met per mm2 was significantly larger; (3) the improvement in LVEF was greater in the Ad5-HGF group than in the mock-vector Ad5 group. We concluded that: (1) high expression of human HGF was observed in the myocardium through intracoronary gene transfection; (2) HGF can improve the mobilization of CD117+/c-Met+ stem cells into ischemic myocardium. The amelioration effect of HGF on postinfarction heart failure could not be limited to stimulating angiogenesis, anti-apoptosis, anti-fibrosis, but was also involved in the recruitment of stem cells into myocardium.

关键词： gene therapy, stem cells, postinfarction heart failure

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Protein Product Encoded by a Human Novel Gene E9730 Enhances AP-1 Activity through Interacting with Jab1

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Acta Biochimica et Biophysica Sinica 2004年第1期36卷 11-15页

作者： Zhao-Qing WANG, Han-Dong WEI, and Fu-Chu HE(department of System Biology, beijing institute of radiation medicine, Chinese Human National Center at beijing, beijing 100850, china) department of System Biology Beijing Institute of Radiation Medicine Chinese Human National Center at Beijing Beijing 100850 China

A novel human gene, named E9730 (a clone number of fetal liver cDNA library), has been identified from more than 14,000 expressed sequence tags (ESTs) based on our large scale sequencing of human fetal liver cDNA libraries. Although sequencing of this novel human gene indicates that it is a leucine zipper protein, the function of E9730 and its homologous genes among species is unknown yet. To find out physiological functional clue of E9730, the yeast two-hybrid system was used to screen the E9730-interac-ting protein(s), and one clone containing a cDNA insert with almost the entire coding sequence (amino acids 39-335) of human Jabl (Jun-activating domain binding protein 1) that interacted specifically with E9730 was identified. A specific association between Jabl and E9730 was shown by co-immunoprecipitation and co-localization experiments. Furthermore, E9730 appeared to enhance Jab 1 -induced AP-1 activity in a concentration-dependent manner and Jabl may be involved in the intracellular signaling trans

关键词： human E9730 gene CSN5/Jabl protein-protein interaction signaling transduction

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Molecular Cloning and Characterization of cDNA Encoding Fibrinolytic Enzyme-3 from Earthworm Eisenia foetida

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Acta Biochimica et Biophysica Sinica 2004年第4期36卷 303-308页

作者： Guo-Qing DONG, Xiao-Ling YUAN, Ya-Jun SHAN, Zhen-Hu ZHAO, Jia-Pei CHEN, and Yu-Wen CONG* department of experimental Therapy of ARS, beijing institute of radiation medicine, beijing 100850, china department of experimental Therapy of ARS Beijing Institute of Radiation MedicineBeijing 100850 China

The earthworm fibrinolytic enzyme-3 (EFE-3, GenBank accession No: AY438622), from the earthworm Eisenia foetida, is a component of earthworm fibrinolytic enzymes. In this study, cDNA encoding the EFE-3 was cloned by RT-PCR. The cDNA contained an open reading frame of 741 nucleotides, which encoded a deduced protein of 247 amino acid residues, including signal sequences. EFE-3 showed a high degree of homology to earthworm (Lumbricus rebullus) proteases F-III-1, F-III-2, and bovine trypsin. The recombinant EFE-3 was expressed in E. coli as inclusion bodies, and the gene encoding the native form of EFE-3 was expressed in COS-7 cells in the medium. Both the refolding product of inclusion bodies and the secreted protease could dissolve the artificial fibrin plate.

关键词： earthworm Eisenia foetida earthworm fibrinolytic enzyme cloning expression refolding

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DNA Damage, Signaling and Repair: Protecting genomic integrity and reducing the risk of human disease

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Chinese Science Bulletin 2011年第30期56卷 3119-3121页

作者： ZHOU PingKun department of radiation Toxicology and Oncology, beijing institute of radiation medicine, beijing 100850, china Special Topic Editor department of radiation Toxicology and Oncology Beijing Institute of Radiation Medicine

DNA is the genetic material that contains the 'instructions' to not only guide the continuation of life but also govern the development, metabolism and functioning of living organisms. DNA damage occurs daily due to v... 详细信息

关键词： DNA损伤基因组伤信号人类疾病整性修复风险保护

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A Putative NES Mediates Cytoplasmic Localization of Apoptin in Normal Cells

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Acta Biochimica et Biophysica Sinica 2004年第12期36卷 817-823页

作者： Qing-Ming WANG*, Guo-Cai FAN, Ji-Zhong CHEN, Hui-Peng CHEN, and Fu-Chu HE* department of Genomics and Proteomics, beijing institute of radiation medicine, beijing 100850, china department of Genomics and Proteomics Beijing Institute of Radiation MedicineBeijing 100850 China

Apoptin, a protein expressed by chicken anemia virus, is found predominantly in the cytoplasm in normal cells, whereas it localizes in the nucleus in transformed and malignant cells. However, the mecha- nisms that regulate the different subcellular localization of Apoptin in normal and tumor cells have not been fully clarified. In this work, a putative nuclear export signal (NES) in Apoptin was predicted. It was testified that the putative NES (pNES) of Apoptin was not a functional NES, but actually acted as a cytoplasmic retention signal. Deletion of the pNES led to the nuclear accumulation of Apoptin in normal cells. In addition, when a strong nuclear localization signal was introduced into Apoptin, it exclusively translocated to the nucleus in normal cells. These observations indicated that the cytoplasmic localization of Apoptin in normal cells results from the balance between cytoplasmic retention and nuclear import. On the other hand, the pNES was also proved to be necessary for Apoptin multimerization. Mutants lacking the pNES did not form obviously visible globular aggregates in normal or tumor cells.

关键词： Apoptin nuclear localization signal cytoplasmic retention signal nuclear export signal

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Properties and applications of embryonic stem cells

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Chinese Science Bulletin 2000年第14期45卷 1258-1265页

作者： GUO Xiaoxia & HE Fuchu1. beijing institute of radiation medicine, beijing 100850, china 2. department of Life Science, Shandong University, Ji’nan 250100, china Correspondence should he addressed to He Fuchu beijing institute of radiation medicine Beijing China department of Life Science Shandong University Ji’nan China

Mouse embryonic stem (ES) cells are pluripotent cells derived from the early embryo and can be propagated stably in undifferentiated state in vitro. They retain the ability to differentiate into all cell types found in the embryonic and adult body in vivo, and can be induced to differentiate into many cell types under appropriate culture conditions in vitro. Using these properties, people have set up various differentiated systems of many cell types and tissues in vitro. Through analysis of these systems, one can identify novel bioactive factors and reveal mechanisms of cell differentiation and organogenesis. ES cell-derived differentiated cells can also be applied to cell transplantation therapy. In addition, we summarized the features and potential applications of human ES cells.

关键词： embryonic stem cell differentiation in vitro cell transplantation therapy gene targeting.

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Detection and quantitation of PS20, a phosphorothioate oligodeoxynucleotides in monkey plasma

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Acta Pharmacologica Sinica 2004年第6期25卷 99-104页

作者： Ming-mei SHANG, Hai-feng SONG2, Xiu-wen LIU, Zhong-ming TANG department of Pharmacology and Toxicology, the institute of radiation medicine, Academy of Military Medical Sciences, beijing 100850, china department of Pharmacology and Toxicology the Institute of Radiation Medicine Academy of Military Medical Sciences

AIM: To establish the method for quantitation of the phosphorothioate oligodeoxynucleotides (S-ODNs) in plasma. METHODS: Two solid-phase extraction columns combined with a strong anion-exchange column were utilized to remove proteins and lipids in plasma, and the salts were removed by a reverse-phase column followed by dialysis with a 2500 Da-cutoff membrane. The concentration of the tested S-ODNs, PS20, and its metabolites extracted from the plasma were determined by the method of non-gel sieving capillary electrophoresis (NGCE) with diode array detection in the presence of internal standard (IS). RESULTS: The method was with good base number specificity. Relative standard deviation % of both intra and inter assay were all less than 10 %, and the total mean recovery was about 91 %. The methodology was successfully used to determine the PK behavior of an anti-tumor antisense S-ODNs in monkeys and identify the metabolites with single base difference. CONCLUSION: The combined method of solid-phase extraction and NGCE could be used to study the pharmacokinetics of S-ODNs, and the main parameters of the methodology met the requirement of PK study.

关键词： antisense oligodeoxyribonucleotides thionucleotides rhesus monkey plasma solid-phase extraction capillary electrophoresis non-gel sieving internal standard

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Therapeutic angiogenesis induced by human hepatocyte growth factor (HGF) gene in rat myocardial ischemia models

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Science Bulletin 2003年第3期48卷 273-276页

作者： WU Danli1*, ZHANG Yourong2*, LAO Miaofen1, QIU Zhaohua1, HA Xiaoqin1 & WU Zuze (WU Cutse)1 1. department of experimental hematology, institute of radiation Medi-cine, beijing100850, china 2. General Hospital of the People抯 Liberation Army of china, beijing 100853, china department of experimental hematology Institute of Radiation Medicine Beijing 100850 China

In order to investigate the feasibility of myo-cardial ischemia gene therapy, we cloned human hepatocyte growth factor gene from human placenta cDNA library by the RT-PCR method. Recombination adenovirus Ad-HGF was constructed by the method of co-transfection and ho-mologous recombination of plasmids in 293 cells. Ad-HGF was amplified in 293 cells and purified through CsCl density gradient centrifugation. Ad-HGF could be expressed in rat primary myocardial cells and HGF secreted into the culture media, which was tested by ELISA. The distribution and persistence of adenovirus in rat were investigated by green fluorescence protein as a report gene. In vivo we found that intramyocardial administration of Ad-HGF could induce angiogenesis in rat myocardium after ligation of coronary artery. The results suggested that Ad-HGF was effective in vitro and in vivo, and the data for designing human trial of gene therapy—— mediated cardiac angiogenesis were pro-vided.

关键词： myocardial ischemia gene therapy recombination ade-novirus hepatocyte growth factor.

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