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Hyaluronan Promotes Tumor Lymphangiogenesis and Intralymphantic Tumor Growth in Xenografts

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Acta Biochimica et Biophysica Sinica 2005年第9期37卷 601-606页

作者： Li-Xia GUO Ke ZOU Ji-Hang JW Hong XIE Laboratory of Biotherapy,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 Laboratory of Biotherapy Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China

Hyaluronan (HA),a high molecular weight glycosaminoglycan in the extracellular matrix,hasbeen implicated in the promotion of malignant phenotypes,including tumor ***,little isknown about the effect of HA on tumor-associated *** this study,mouse hepatocellularcarcinoma Hca-F cells combined with or without HA were injected subcutaneously into C3H/Hej mice,thenangiogenesis and lymphangiogenesis of implanted tumors were examined by immunostaining for platelet-endothelial cell adhesion molecule-1 and lymphatic vascular endothelial hyaluronan receptor-1 ***,we found HA promotes tumor lymphangiogenesis and the occurrence of intratumoral lymphaticvessels,but has little effect on tumor ***,HA also promotes intralymphatic tumorgrowth,although it is not sufficient to potentiate lymphatic *** results suggest that HA,which is elevated in most malignant tumor stroma,may also play a role in tumor progression by promotinglymphangiogenesis.

关键词： hyaluronan tumor lymphangiogenesis intralymphatic tumor growth

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 SciencesShanghai 200031 China

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

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 SciencesShanghai 200031 China

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

Detection of Tumor Marker CA125 in Ovarian Carcinoma Using Quantum Dots

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Acta Biochimica et Biophysica Sinica 2004年第10期36卷 681-686页

作者： Hui-Zhi WANG, Hai-Yan WANG, Ru-Qiang LIANG, and Kang-Cheng RUAN* Key Laboratory of Proteomics, 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 Key Laboratory of Proteomics Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China

Semiconductor quantum dots (QDs) offer several advantages over organic dyes in fluores- cence-imaging applications, such as higher quantum yield, exceptional photostability, and a narrow, tunable, and symmetric emission spectrum. To explore whether QDs could specifically and effectively label tumor markers and be used in immunohistochemistry as a novel type of fluorescent probe, we used quantum dots with maximum emission wavelength 605 nm (QD605) to detect the ovarian carcinoma marker CA125 in specimens of different types (fixed cells, tissue sections, and xenograft piece). Additionally, we compared the photostability of QD signals with that of a conventional organic dye, FITC. All labeling signals of QDs were found to be more specific and brighter than those of FITC. Moreover, the QDs exhibited exceptional photostability during continuous illumination for 1 h by a high-intensity laser (Ar laser power 100 mW) at 488 nm, while the FITC signals faded very quickly and became undetectable after 24 min of illumination. These results indicate that QD-based probes can offer substantial advantages over existing fluorophores in many applications, and can be used effectively in immunohistochemistry as a novel class of fluorescent probes.

关键词： quantum dots detection of tumor marker CA125 ovarian carcinoma

Rational redesign of inhibitors of furin/kexin processing proteases by electrostatic mutations

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Acta Pharmacologica Sinica 2004年第12期25卷 146-152页

作者： Xiao-hui CAI, Qing ZHANG, Da-fu DING2 Key Laboratory of Proteomics, 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 中科院上海生命科学院 Key Laboratory of Proteomics CAS 上海 200031

AIM: To model the three-dimensional structure and investigatethe interaction mechanism ofthe proprotein convertase furin/kexin and their inhibitors (eglin c mutants). METHODS: The three-dimensional complex structures of furin/ kexin with its inhibitors, eglin c mutants, were generated by modeller program using the newly published X-ray crystallographical structures of mouse furin and yeast kexin as templates. The electrostatic interaction energy of each complex was calculated and the results were compared with the experimentally determined inhibition con- stants to find the correlation between them. RESULTS: High quality models of furin/kexin-eglin c mutants were obtained and used for calculation of the electrostatic interaction energies between the proteases and their inhibitors. The calculated electrostaticenergies of interaction showeda linear correlation to the experimental inhibition constants. CONCLUSION: The modeled structures give good explanations of the specificity of eglin c mutants to furin/ kexin. The electrostatic interactions play important roles in inhibitory activity of eglin c mutants to furin/kexin. The results presented here provided quantitative structural and functional information concerning the role of the charge- charge interactions in the binding of furin/kexin and their inhibitors.

关键词： furin kexin eglin c drug design electrostatic mutation

Multiple Signaling Pathways Control Tbx6 Expression during Xenopus Myogenesis

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Acta Biochimica et Biophysica Sinica 2004年第6期36卷 390-396页

作者： Pan-Feng FANG, Rui-Ying HU, Xing-Yue HE, and Xiao-Yan DING* Laboratory of Molecular and cell biology, Laboratory of Stem cell 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 Laboratory of Molecular and cell biology Laboratory of Stem Cell Biology & Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China

Tbx6 is critical for somite specification and myogenesis initiation. It has been shown that Activin/Nodal, VegT/Nodal, FGF, and BMP signaling pathways are involved early in specifying mesoderm or later in patterning mesoderm, and Xnot plays roles in setting up the boundary between notochord and paraxial mesoderm. In this study, we introduce the dominant negative form of above genes into embryos to evaluate if they are responsible for regulating Tbx6 expression. The results show that: (1) Activin/Nodal and VegT/Nodal signals are necessary for both initiation and maintenance of Tbx6 expression, and Nodal is sufficient to induce ectopic Tbx6 expression; (2) FGF signal is necessary for the initiation and maintenance of Tbx6, but it is not sufficient to induce Tbx6 expression; (3) BMP is also necessary for the expression of Tbx6, and the induction of Tbx6 expression by BMP is dose dependent; (4) Xnot has no effect on the expression of Tbx6. Our results suggest that several signaling pathways are involved in regulating Tbx6 expression, and pave the route to reveal the molecular mechanism of initiating myogenesis.

关键词： Tbx6 FGF BMP Activin/Nodal VegT/Nodal

Association between Alcohol, Smoking and HLA-DQA1*0201 Genotype in Psoriasis

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Acta Biochimica et Biophysica Sinica 2004年第9期36卷 597-602页

作者： Guang-Yong ZHENG1,2#, Sheng-Cai WEI3#, Tie-Liu SHI1, and Yi-Xue LI1* 1Bioinformatics Center, shanghai institutes for biological sciences, chinese academy of sciences, graduate school of the chinese academy of sciences, shanghai 200031, china Research Center for Proteome Analysis, 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 3Guangzhou institute of Dermatology, Guangzhou 510095, china Bioinformatics Center Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China Research Center for Proteome Analysis Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China Guangzhou institute of DermatologyGuangzhou 510095 China

Psoriasis is a chronic skin disease triggered by genetic, environment or other risk factors such as infection, drugs, stress, moisture, alcohol, and smoking. A major psoriasis susceptibility locus at 6p21.3 has been identified. Further studies found that HLA-DQA1*0201 allele was associated with psoriasis. However, there were few data exploring an association between the environmental factors and susceptibility genes. In this study, the samples of 189 patients with psoriasis and 333 healthy controls were collected with their consent and were carried on analysis through polymerase chain reaction sequence-specific primer (PCR- SSP) method. The proportion of male psoriasis patients engaging in the smoking and alcohol was much higher than that of the control group (P6.91, P<1.0×10–4).

关键词： psoriasis HLA-DQA1*0201 allele smoking alcohol association

C/EBPαp30 plays transcriptional regulatory roles distinct from C/EBPαp42

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cell Research 2007年第4期17卷 374-383页

作者： Chunxi Wang~1 Xiaotao Chen~1 Yanping Wang~1 Jialei Gong~1 Gengxi Hu ~1State 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 StateKeyLaboratoryofMolecularbiology InstituteofBiochemistryandCellBiologyShanghaiInstitutesforBiologicalSciencesChineseAcademyofSciencesGraduateSchooloftheChineseAcademyofSciencesShanghai200031China

CCAAT/enhancer binding protein α(C/EBPα)is a transcriptional regulatory factor that inhibits cell proliferation,andalternative translational initiation produces two polypeptides,C/EBPcαp30 and C/EBPα*** expression profiling,it was revealed that C/EBPαp30 specifically inhibited a unique set of genes,including MPP11,p84N5 and SMYD2,which were not affected by C/EBPαp42 in both QSG-7701 hepatocyte cell line and QGY-7703 hepatoma ***-quantitative RT-PCR analysis independently confirmed these *** immunoprecipitation assay showed thatC/EBPαp30 bound to the promoters of these genes more strongly than C/EBPα*** clinical hepatoma samples inwhich C/EBPα was downregulated,all three genes specifically inhibited by C/EBPαp30 were ***,repression of MPP11,p84N5 and SMYD2 genes might not be directly involved in C/EBPαp30-mediated growth *** data suggest that C/EBPαp30 regulates a unique set of target genes and is more than a dominant-negativeregulator of C/EBPαp42.

关键词： C/EBPα expression pattern p84N5 MPP11 SMYD2

Synthesis and Characteristics of an Aspartame Analogue,L-Asparaginyl L-3-Phenyllactic Acid Methyl Ester

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Acta Biochimica et Biophysica Sinica 2004年第6期36卷 385-389页

作者： Hu TAO1, Da-Fu CUI1, and You-Shang ZHANG1,2* 1institute 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 institute of Protein Research, Tongji University, shanghai 200031, china institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China institute of Protein Research Tongji UniversityShanghai 200031 China

An aspartame analogue, L-asparaginyl L-3-phenyllactic acid methyl ester was synthesized with aspartic acid replaced by asparagine and peptide bond replaced by ester bond. The aspartic acid of aspartame could be replaced by asparagine as reported in the literature. In this analogue, the hydrogen of amide group could still form a hydrogen bond with the oxygen of ester bond and the ester bond was isosteric with peptide bond. However, the product was not sweet, showing that the peptide bond could not be replaced by ester bond. The peptide C-N bond behaves as a double bond that is not free to rotate and the C, O, N and H atoms are in the same plane. The replacement of peptide bond by ester bond destroyed the unique conformation of peptide bond, resulting in the loss of sweet taste.

关键词： artificial sweetener aspartame derivative peptide bond