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cell Research 2001年第2期11卷 85-88页

作者： GENG JIAN Guo institute of biochemistry and cell biology, Shanghai institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue Yang Road, Shanghai 200031, China institute of biochemistry and cell biology Chinese Academy of Sciences Shanghai. jggeng@***

Directional migration of leukocytes is indispensable to innate immunity for host defense. However, recruitment of leukocytes to a site of tissue injury the constitutes a leading cause for inflammatory responses. Mechanistically, it involves a cascade of cellular events precisely regulated by temporal and spatial present tation of a repertoire of molecules in the migrating leukocytes and their surroundings (microenvironments). Here I will summarize the emerging evidence that has shed lights on the underlying molecular mechanism for directional migration of leukocytes, which has guided the therapeutical development for innovative anti-inflammatory medicines.

关键词： Directional cell migration leukocytes inflammation therapy.

An Effective Method for Raising Antisera Against β-defensins: Double-copy Protein Expression of mBin1b in E. coli

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Acta Biochimica et Biophysica Sinica 2004年第8期36卷 571-576页

作者： Li-Qing XIAO, Ai-Hua LIU, and Yong-Lian ZHANG State Key Laboratory of Molecular biology, institute of biochemistry and cell biology, Shanghai institutes for Biological Sciences, Chinese Academy of Sciences, Graduate School of the Chinese Academy of Sciences, Shanghai 200031, China State Key Laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of Sciences

Bin1b is a rat epididymis specific β-defensin which may have fertility related functions in addition to its antimicrobial activity. β-defensins are cysteine-rich cationic antimicrobial peptides that have their important implications in innate and adaptive immunity. Though considerable numbers of new β-defensins have been discovered, few corresponding antibodies have been reported. The small peptide with special structure and antimicrobial nature of β-defensins make them very difficult to express in prokaryotic system. Here we adopted a double-copy protein expression scheme based on which not only the mBin1b protein was successfully expressed but also the immunity of the antigen was enhanced. The validity of the antisera was verified by using Western blotting and immunohistochemical analyses. It will be a useful tool for deeply investigating the roles of Bin1b and also provide a simple but effective method in raising antisera against other members of the β-defensin gene family.

关键词： Bin1b epididymis β-defensin double-copy protein polyclonal antiserum

Functional Analysis of Multiple Transcription Factor Sites in a Regulatory Element of Human ε-Globin Gene

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Acta Biochimica et Biophysica Sinica 2004年第10期36卷 673-680页

作者： Chun-Hui HOU, Jian HUANG, and Ruo-Lan QIAN* State Key Laboratory of Molecular biology, institute of biochemistry and cell biology, Shanghai institutes for Biological Sciences, Chinese Academy of Sciences, Graduate School of the Chinese Academy of Sciences, Shanghai 200031, China State Key Laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of Sciences

The developmental control of the human ε-globin gene expression is mediated by transcrip- tional regulatory elements in the 5’ flanking DNA of this gene. A previously identified negative regulatory element (–3028 to –2902 bp, termed ε-NRAII) was analyzed and one putative NF-κB site and two GATA sites locate at –3004 bp, –2975 bp and –2948 bp were characterized. Electrophoresis mobility shift assay (EMSA) showed that the putative NF-κB site was specifically bound by nuclear proteins of K562 cells. Data obtained from transient transfection showed that the expression of reporter gene could be upregulated about 50% or 100% respectively when ε-NRAII was inserted upstream of the SV40 promoter or ε-globin gene proximal promoter (–177 bp to +1 bp), suggesting that ε-NRAII might not be a classic silencer. Mutation in the putative NF-κB site or in the GATA site (at –2975 bp) slightly reduced the expression of reporter gene driven by SV40 promoter or ε-globin gene proximal promoter. However, the mutation of GATA site at –2948 bp remarkably reduced the reporter gene activity driven by SV40 promoter, but not by ε-globin gene proximal promoter. Further mutation analysis showed that the negative effect of mutation in GATA site at –2948 bp on SV40 promoter was not affected by the mutation of the putative NF-κB site, whereas it could be abolished by the mutation of GATA site at –2975 bp. Furthermore, the mutation of both GATA sites could synergistically reduce the reporter gene activity driven by ε-globin gene proximal promoter. Those results suggested that ε- NRAII might function differently on the SV40 promoter and ε-globin gene proximal promoter.

关键词： ε-globin gene regulatory element NF-κB GATA

Bcl-2 over-expression and activation of protein kinase C suppress the Trail-induced apoptosis in Jurkat T cells

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cell Research 2001年第2期11卷 101-106页

作者： GUO BEN CHANG, YONG HUA XU institute of biochemistry and cell biology, Shanghai institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue-yang Road, Shanghai 200031, China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue-yang Road Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue-yang Road Shanghai 200031 China

Trail, a tumor necrosis factor-related apoptosis-inducing ligand, is a novel potent endogenous activator of the cell death pathway through the activation of cell surface death receptors Trail-R1 and Trail-R2. Its role, like FasL in activation-induced cell death (AICD), has been demonstrated in immune system. However the mechanism of Trail induced apoptosis remains nuclear. In this report, the recombinant Trail protein was expressed and purified. The apoptosis-inducing activity and the regulation mechanism of recombinant Trail on Jurkat T cells were explored in vitro. Trypan blue exclusion assay demonstrated that the recombinant Trail protein actively killed Jurkat T cells in a dose-dependent manner. Trail-induced apoptosis in Jurkat T cells were remarkably reduced by Bcl-2 over expression in BcL-2 gene transfected cells. Treatment with PMA (phorbol 12-myristate 13-acetate), a PKC activator, suppressed nail-induced apoptosis in Jurkat T cells. The inhibition of apoptosis by PMA was abolished by pretreatment with Bis, a PKC inhibitor. Taken together, it was suggested that Bcl-2 over-expression and PMA activated PKC actively down-regulated the Trail-mediated apoptosis in Jurkat T cell.

关键词： trail Jurkat T cells apoptosis Bcl-2 PEA PKC Bis.

MicroRNA in cell differentiation and development

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Science China(Life Sciences) 2009年第3期52卷 205-211页

作者： SHI Yi & JIN YouXin State Key Laboratory of Molecular biology,Shanghai institute of biochemistry and cell biology,Shanghai institutes for Biological Sciences,Chinese Academy of Sciences,Shanghai 200031,China State Key Laboratory of Molecular biology Shanghai Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

The regulation of gene expression by microRNAs(miRNAs) is a recently discovered pattern of gene regulation in animals and *** have been implicated in various aspects of animal development and cell differentiation,such as early embryonic development,neuronal development,muscle development,and lymphocyte development,by the analysis of genetic deletions of individual miRNAs in *** studies show that miRNAs are key regulators in animal development and are potential causes of human *** we review some recent discoveries about the functions of miRNAs in cell differentiation and development.

关键词： miRNA embryonic development neuronal development muscle development lymphocyte development

维普期刊数据库

Identification of differentially expressed genes in human uterine leiomyomas using differential display

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cell Research 2002年第1期12卷 39-45页

作者： BIN LI, MEI SUN, BIN HE, JIN YU, You DUAN ZHANG, YONG LIAN ZHANG, State Key Laboratory of Molecular biology, institute of biochemistry and cell biology, Shanghai institute for Biological Science, Chinese Academy of Sciences, 820, Yue- Yang Road, Shanghai, 1State Key Laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institute for Biological Science Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China 1State Key Laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institute for Biological Science Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China 1State Key Laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institute for Biological Science Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China 2OB/GYN Hospital Shanghai (First) Medical University 419 Fang-Xie Road Shanghai China 1State Key Laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institute for Biological Science Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China 1State Key Laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institute for Biological Science Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China

In searching of differentially expressed genes in human uterine leiomyomas, differential display was used with twelve pairs of primers to compare human uterine leiomyomas with matched myometrium. False positives were eliminated by reverse Northern analysis. Positives were confirmed by Northern blot analysis. RESULTS: [1] Four of 69 cDNA fragments (3 up-regulated named L1, L2 and L3 and 1 down-regulated named Mi in leiomyoma) were confirmed by Northern analysis. [2] Sequence comparison and Northern analysis proved that Li is exactly the human ribosomal protein Si9. [3] It was present ubiquitously in i3 tissues tested but in various levels and even in different size. [4] Li was highly expressed in parotidean cystadenocarcinoma, pancreatic cancer and breast cancer examined. [5] No mutations have been found in human uterine leiomyomas (n=6). CONCLUSIONS: hRPSi9 overexpression might be a universal signal in rapid cell growth tissues.

关键词： Human uterine leiomyomas differentially expressed gene differential display (DD) human S19 ribosomal protein (hRPS19) up-regulated gene in human leiomyomas.

Mechanisms of microRNA-mediated gene regulation

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Science China(Life Sciences) 2009年第12期52卷 1111-1116页

作者： ZHAO Shuang & LIU Mo-Fang· State Key Laboratory of Molecular biology, institute of biochemistry and cell biology, Shanghai institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200031, China State Key Laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences the Chinese Academy of Sciences Shanghai 200031 China

microRNAs (miRNAs) are identified as a class of non-protein regulators and a new source for broad control of gene expression in eukaryotes. The past years have witnessed substantial progress in understanding miRNA functions and mechanisms, although a few controversies remain. Various hypotheses and models have been suggested for the mechanisms of miRNA repression, including translational inhibition at the level of initiation or elongation, rapid degradation of the nascent peptide, mRNA degradation, and mRNA sequestration into P bodies (processing bodies) and SGs (stress granules) for degradation or/and storage. Recently, some noncanonical miRNA regulation, such as miRNA activation and de-repression of miRNA inhibition, have been uncovered. This review discusses some recent advances about how miRNAs regulate their targets and various modes of miRNA function.

关键词： miRNA repression activation de-repression processing bodies stress granules

维普期刊数据库

Interaction of vascular endothelial cells with leukocytes, platelets and cancer cells in inflammation, thrombosis and cancer growth and metastasis

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Acta Pharmacologica Sinica 2003年第12期 123-126页

作者： GENG Jian-Guo institute of biochemistry and cell biology, Shanghai institutes for Biological Sciences Chinese Academy of Sciences, Shanghai 200031, China

Adhesion and migration of mammalian cells are of crucial importance in a number of biological events, such as fertilization, embryogenesis, pattern, tissue and organ formation, and in a variety of physiological and pathological processes, including lymphocyte trafficking, leukocyte recruitment, hemostasis, wound healing, tumor angiogenesis and cancer metastasis. All these

关键词： of et as in Interaction of vascular endothelial cells with leukocytes thrombosis and cancer growth and metastasis with

Adenovirus-mediated expression of pig α(1,3) galactosyltransferase reconstructs Gal α(1,3) Gal epitope on the surface of human tumor cells

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cell Research 2001年第2期11卷 116-124页

作者： XING LI, GUO HONG XIA, JIAN FEI, FANG HUANG, LI HE GUO institute of biochemistry and cell biology, Shanghai institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

Gal α(1, 3) Gal (gal epitope) is a carbohydrate epitope and synthesized in large amount by α(1, 3) galactosyltransferase [α(1, 3) GT] enzyme on the cells of lower mammalian animals such as pigs and mice. Human has no gal epitope due to the inactivation of α(1, 3) GT gene but produces a large amount of antibodies (anti-Gal) which recognize Gal α(1, 3) Gal structures specifically. In this study, a replication deficient recombinant adenoviral vector Ad5sGT containing pig α(1, 3) GT cDNA was constructed and characterized. Adenoviral vector-mediated transfer of pig α(1, 3) GT gene into human tumor cells such as malignant melanoma A375, stomach cancer SGC-7901, and lung cancer SPC-A-1 was reported for the first time. Results showed that Gal epitope did not increase the sensitivity of human tumor cells to human complement-mediated lysis, although human complement activation and the binding of human IgG and IgM natural antibodies to human tumor cells were enhanced significantly after Ad5sGT transduction. Appearance of gal epitope on the human tumor cells changed the expression of cell surface carbohydrates reacting with Ulex europaeus I (UEA I) lectins, Vicia villosa agglutinin (VVA), Arachis hypogaea agglutinin (PNA), and Glycine max agglutinin (SBA) to different degrees. In addition, no effect of gal epitope on the growth in vitro of human tumor cells was observed in MTT assay.

关键词： Adenoviral vector galactosyltransferase Gal α(1 3) Gal gene expression human tumor cell.

Overexpression of a novel gene , Cms1, can rescue the growth arrest of a Saccharomyces cerevisiae mcm10 suppressor

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cell Research 2001年第4期11卷 285-291页

作者： WANG JI WU, JIA RUI WU State Key Laboratory of Molecular biology, institute of biochemistry and cell biology, Shanghai Research Center of Life Science, Shanghai institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yue- Yang Road, Shanghai 1State Key Laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Research Center of Life Science Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China 1State Key Laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Research Center of Life Science Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China

MCM10 protein is an essential replication factor involved in the initiation of DNA replication. A mcm10 mutan (mcm10-1) of budding yeast shows a growth arrest at 37 C. In the present work, we have isolated a mcm10-1 suppressor strain, which grows at 37 C. Interestingly, this mcm10-1 suppressor undergoes cell Cycle arrest at 14 C. A novel gene, YLR003c, is identified by high-copy complementation of this suppressor. We called it as Cms1 (Complementation of Mcm 10 Suppressor). Furthermore, the experiments of trans- formation show that cells of mcm10-1 suppressor with high-copy plasmid but not low-copy plasmid grow at 14 C, indicating that overexpression of Cms1 can rescue the growth arrest of this mcm10 suppressor at non-permissive temperature. These results suggest that CMS1 protein may functionally interact with MCM10 protein and play a role in the regulation of DNA replication and cell cycle control.

关键词： Saccharomyces cerevisiae DNA replication Mcm10 Cms1 cell cycle.