Cell Adhesion Molecules ; metabolism ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; physiology ; Humans ; Integrins ; metabolism ; Membrane Proteins ; metabolism ; Neoplasms ; metabolism ; pathology ; Organ Specificity ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Saphenous Vein ; cytology ; Tumor Cells, Cultured ; Umbilical Veins ; cytology

In the post-genomic era, with the accomplishment of the sequence mapping of human genome, one of the most important tasks for life science is the explanation and identification of human genome, that is, about 1/3 genes of human genome and their functions need further revealment and verification on the level of protein. In the field of functional proteomics, the human disease proteomics shows great potential in the discovery of new molecular targets and biomarkers for medicine and biopharmacy. In this article, we have made a concise discussion on the current status, existing problems and future development in the research of human disease proteomics both in and out of China.
Biomarkers ; Genomics ; Humans ; Proteomics ; trends

Scansite is a short linear motif-based scanning approach established in the latest two years. It's accessible over the World Wide Web and can be used to identify sequence motifs likely to be phosphorylated by specific protein kinases or likely to bind to specific protein domains such as 14-3-3, SH2 and SH3 domains. The usage and function of the potent approach were reviewed and compared with previously established tools for phosphorylation prediction. The facing problems and application outlook of Scansite in prediction of cell signaling networks within proteomes were also presented.
Amino Acid Motifs ; Amino Acid Sequence ; Databases, Factual ; Internet ; Molecular Sequence Data ; Phosphorylation ; Protein Structure, Tertiary ; Proteins ; chemistry ; metabolism ; Signal Transduction

Liver Diseases ; immunology ; metabolism ; Signal Transduction ; Toll-Like Receptors ; immunology ; metabolism

OBJECTIVETo study the metastasis-associated molecules differentially expressed in highly and poorly metastatic sublines and the mechanism of metastasis in lung giant cell carcinoma.

METHODSHighly and poorly metastatic sublines (PLA801D and PLA801C)were used as metastasis model. Cell motility and invasion assay in vitro were first compared between the two sublines. Then, gelatin zymography analysis was used to determine the MMP-2 and MMP-9 activity. The protein expression level of secreted MMP-2, MMP-9, TIMP-1, TIMP-2 and intracellular expression level of p53, p16, PCNA, CD44(V6) isomeride, E-cadherin, CK18, nm23-H1 as well as the mRNA expression level of MMP-2, MMP-9, TIMP-1, TIMP-2, VEGF were compared through Western blot. Semi-quantitative RT-PCR analysis was used to determine the intracellular mRNA expression of MMP-2, MMP-9, TIMP-1, TIMP-2 and VEGF.

RESULTSThe in vitro cell invasion potential of highly metastatic subline PLA801D was significantly higher than that of poorly metastatic subline PLA801C by about 4 folds, while the cell motility potential was similar. The secreted MMP-2 activity was notably higher in PLA801D, which was initiated by the higher expression of MMP-2 at protein and mRNA level. In addition, the expression level of p53, PCNA, CK18 protein and VEGF mRNA were significantly higher, while the expression level of p16, E-cadherin and nm23-H1 protein were significantly lower in PLA801D. Some molecules such as MMP-9, TIMP-1, TIMP-2, CD44(V6) isomeride, which had been reported to be associated with tumor metastasis, were not observed to change significantly between the two sublines.

CONCLUSIONThere are significant differences in metastatic potential and phenotypes between highly and poorly metastatic sublines of lung giant cell carcinoma. Some differentially expressed molecules might be playing roles in promoting or inhibiting metastasis of lung giant cell carcinoma, which may be useful to elucidate the mechanism of metastasis.

Carcinoma, Giant Cell ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Interleukin-8 ; genetics ; Lung Neoplasms ; metabolism ; pathology ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Neoplasm Invasiveness ; Neoplasm Metastasis ; RNA, Messenger ; analysis ; Tissue Inhibitor of Metalloproteinase-1 ; analysis ; Vascular Endothelial Growth Factor A ; genetics

Objective： The current study was designed to clarify whether hepatopoietin (HPO) stimulates autonomous growth of hepatoma cell by autocrine loop. Methods： The authors conducted experiments in vitro with hepatoma cell lines. RT-PCR, ELISA and Western blot were used to examine HPO expression in hepatoma cells. Blocking effect of HPO by HPO neutralizing antibody was utilized and the changes of cell proliferation was observed. Results： HPO was expressed by hepatoma cells and secreted into the medium. Moreover, the HPO antibody inhibited specifically the autonomous proliferation of hepatoma cell and antagonized the stimulatory effect of concentrated conditioned medium derived from hepatoma cell HepG2. Conclusion： The results strongly suggest that HPO acts as an autocrine factor to maintain the autonomous growth of hepatoma cells.

Fatty Liver ; immunology ; Immune Tolerance ; Liver ; immunology

Hepatic Stellate Cells ; immunology

The human basic Krüppel-like factor (hBKLF) is a newly cloned human transcription factor from the cDNA library of fetal liver. It belongs to the Krüppel-like transcription factor family. Previous expression study showed that it is a hematopoietic related factor. This study was aimed to investigate the effect of hBKLF on cell proliferation, differentiation and hemoglobin synthesis by using K562 cell line as model. The sense and antisense expression plasmids of hBKLF were constructed, and transfected into K562 cells by lipofectamine. After G418 selection for 4 weeks, the cell line with stable expression of the gene was obtained. Then the hBKLF expression level, proliferation ability, colony formation and hemoglobin production were detected by RT-PCR and Western blot, MTT method, methyl cellulose semisolid culture method and benzidine test respectively. The morphologic change of cell was observed with inverted microscope. The results showed that the sense plasmid could increase hBKLF level and antisense plasmid could decrease hBKLF expression. When hBKLF level was down-regulated, K562 cells could proliferate more quickly and synthesize more hemoglobin. But there were no differences in colony formation ability and no apparent morphologic change. It is concluded that hBKLF can inhibit hematopoietic cell proliferation and hemoglobin synthesis. It is suggested that hBKLF plays an important role in the proliferation and differentiation of hematopoietic cells.
Animals ; COS Cells ; Cell Differentiation ; physiology ; Cell Proliferation ; drug effects ; Cell Transformation, Neoplastic ; drug effects ; Cercopithecus aethiops ; Hemoglobins ; biosynthesis ; Humans ; K562 Cells ; Kruppel-Like Transcription Factors ; biosynthesis ; genetics ; pharmacology ; Transcription Factors ; biosynthesis ; genetics ; Transfection

Fatty Liver ; Humans ; Proteomics ; methods