ABSTRACT
Objective:To study the differentially expressed proteins in saliva of health people and the patients with oral squamous cell carcinoma(OSCC).Methods:Saliva of 17 cases with OSCC and paired health subjects was collected,the proteins in the saliva were examined by two-dimensional gel electrophoresis (2-DE) separation,the proteins were examined by 2-DE separation,the saliva proteome dimensional electrophoresis profiles were obtained by MALDI-TOF/MS mass spectrometric identification,the information of the differentially expressed protein in OSCC group was studied by NCBI database bioinformatics analysis.Results:10 proteins differentially expressed between the 2 groups were observed by mass spectrometry.Bioinformatics analysis showed that S100A8,S100A8/S100A9 and Epidermal cytokeratin 2(EK2) were highly expressed in the saliva of OSCC cases.Conclusion:S100A8,S100A8/S100A9 and EK2 may be related to the development of OSCC.
ABSTRACT
Gamma-tocotrienol (GTT) has been shown to exhibit significant antitumor activity in a variety of tumor cells. Previous findings have demonstrated that GTT had antiprolifera-tive effects on a liver cancer cell line (HepG2) with an IC50 value of 170μM. In this study, two dimensional gel electrophoresis (2DE) was used to determine changes in protein expression in HepG2 cell line following treatment with GTT. The ultimate aim is to identify the possible molecular mechanisms involved in GTT antitumor activity. This study is focused on obtaining a 2DE protein profile for HepG2 cell line with and without GTT treatment. In the preliminary analysis of the resulting 2DE profiles, 18 protein spots were found to be differentially expressed in cells treated with GTT. This observation is confirmed by extending the analysis to a larger sample size. By studying the effects of GTT treatment on differential protein expression in HepG2 cells the underly-ing mechanisms involved in the antitumor activity of GTT may be elucidated.
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OBJECTIVE: Comparison of protein expressions by two-dimensional gel electrophoresis (2-DE) in normal cervix and squamous cell carcinoma tissues in Korean women. METHODS: Normal cervix and squamous cell carcinoma tissues were solubilized with 2-DE buffer and the first dimension of PROTEAN IEF CELL, isoelectric focusing (IEF), was performed using pH3-10 linear IPG strips of 17 cm. And then running 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and sliver stain. Scanned image was analyzed using PDQuest 2-D softwareTM. Protein spot spectrum was identified by assisted laser desorption/ionization-time of fighting (MALDI-TOF) and the protein mass spectrum identifications were performed by searching protein databases of Swiss-prot/TrEMBL, Mascot and MS-FIT. RESULTS: We found 9 up-regulation proteins (Alpha enolase, Keratin 19 type I, Keratin 20 type I, Keratin 13 type I, beta-actin, Aflatoxin B1 aldehyde reductase 1, Annexin A2, Squamous cell carcinoma antigen 2, unknown), 7 down-reguation proteins (Annexin 1, Myosin regulatory light chain 2, 14-3-3 protein epsilon, Heat shock 27 kDa protein, Hypothetical protein (DKFZP434C1715), Tumor necrosis factor receptor superfamily member 13B, Smoth muscle protein 22-alpha) and 6 up and down-regulation proteins (Tropomyosin 1, Tropomyosin 2, Tropomyosin 3, Serine (or cysteine) proteinase inhibitor, Phosphatidylinositol transfer protein alpha isoform, Src homology 3 domain-containing protein HIP-55) between normal cervix and squamous cell carcinoma cell tissues. CONCLUSION: 2-DE offers total protein expressions between normal cervix and squamous cell carcinoma cell tissues, and searching of differently expressed protein for the diagnostic markers of squamous cell carcinoma tissue.
Subject(s)
Female , Humans , 14-3-3 Proteins , Actins , Aflatoxin B1 , Aldehyde Reductase , Annexin A2 , Carcinoma, Squamous Cell , Cervix Uteri , Databases, Protein , Down-Regulation , Electrophoresis, Gel, Two-Dimensional , Electrophoresis, Polyacrylamide Gel , Hot Temperature , Isoelectric Focusing , Keratin-13 , Keratin-19 , Keratin-20 , Mass Spectrometry , Muscle Proteins , Myosin Light Chains , Phospholipid Transfer Proteins , Phosphopyruvate Hydratase , Receptors, Tumor Necrosis Factor , Running , Serine , Shock , Sodium Dodecyl Sulfate , Tropomyosin , Up-Regulation , Uterine Cervical NeoplasmsABSTRACT
OBJECTIVE: Comparison of protein expression by two-dimensional gel electrophoresis (2-DE) in normal myometrium and uterine leiomyoma in Korean women. METHODS: Normal myometrium and uterine leiomyoma tissues were solubilized with 2-DE buffer and the first dimension of PROTEAN IEF CELL, isoelectric focusing (IEF), was performed using pH4-8 linear IPG strips of 17 cm. And then running 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS- PAGE) and sliver stain. Scanned image analyzed using PDQuest 2-D softwareTM. Protein spot spectrum was identified by assisted laser desorption/ionization-time of fighting (MALDI-TOF) and the protein mass spectrums identification were performed by searching protein databases of Swiss-prot/TrEMBL, Mascot and MS-FIT. RESULTS: In this study, we found 17 up-regulated proteins (phosphate carrier protein, 60 kDa heat shock protein, acidic calcium-independent, glutathione transferase omega, chloride intracellular channel 4, Ras-related protein Rab-11B, phosphatidylinositol transfer protein alpha isoform, type II keratin subunit protein, Cofilin 2 isoform 1, transgelin, ATP carrier protein, alpha-catenin homolog, parkinson disease 2, apo-cellular retinoic acid binding protein II, osteoglycin preproprotein, proteasome activator subunit 1 isoform, Unnamed protein) and 7 down-regulated proteins (Serum amyloid P component, annexin IV, alpha 1 actin precursor, hypoxanthine-guanine phosphoribosyltransferase, tumor necrosis factor receptor superfamily member EDAR precursor, peroxiredoxin 2, translation elongation factor EF-Tu precursor) between myometrium and leiomyoma. CONCLUSION: 2-DE offer total protein expression between normal myometrium and uterine leiomyoma, and searching of differently expressed protein for the diagnostic markers of leiomyoma.
Subject(s)
Animals , Female , Humans , Mice , Actins , Adenosine Triphosphate , alpha Catenin , Annexin A4 , Carrier Proteins , Cofilin 2 , Databases, Protein , Electrophoresis, Gel, Two-Dimensional , Electrophoresis, Polyacrylamide Gel , Glutathione Transferase , Heat-Shock Proteins , Hypoxanthine Phosphoribosyltransferase , Isoelectric Focusing , Keratins, Type II , Leiomyoma , Myometrium , Parkinsonian Disorders , Peptide Elongation Factor Tu , Peptide Elongation Factors , Peroxiredoxins , Phospholipid Transfer Proteins , Proteasome Endopeptidase Complex , Receptors, Tumor Necrosis Factor , Running , Serum Amyloid P-Component , Sodium Dodecyl Sulfate , TretinoinABSTRACT
Identification of expressed protein profiles and antigenic determination are some of the most challenging aspects of proteomics. Two-dimensional gel electrophoresis (2-DE) combined with immunoblot analysis were employed to study the N. caninum proteome. Protein sample preparation was carried out by first conducting sonication, followed by adding lysis buffer containing 7M urea plus 2M thiourea to the purified tachyzoites in order to complete disruption. A total of 335 differentially expressed protein spots were detected using pH 4-7 IPG strip (7 cm) that were run in a 56 kVh isoelectric focusing (IEF) system. Of the spots analyzed, 64 were identified as antigenic spots on immunoblot profile. Major antigenic spots appeared at 65 kDa (pI 5.2-5.3), 51 kDa (pI 5.5), 38 kDa (pI 5.1), 33 kDa (pI 4.4), 29 kDa (pI 5.6) and 15.5 kDa (pI 5.0) were observed to be significantly distinct compared to the rest of the antigenic spots. The results indicate that combination of 2-DE and immunoblotting methods were thought as very useful tools in defining both proteins and antigens of N. caninum tachyzoites. Additionally, present 2-DE profiles may be valuable in further proteomic approaches and study of the pathogen.
Subject(s)
Animals , Antibody Formation , Antigens, Protozoan/analysis , Electrophoresis, Gel, Two-Dimensional/methods , Electrophoresis, Polyacrylamide Gel , Epitopes/analysis , Image Processing, Computer-Assisted , Immunoblotting/methods , Isoelectric Focusing , Neospora/chemistry , Proteome/analysis , Proteomics , Protozoan Proteins/analysisABSTRACT
Objective:To establish two-dimensional electrophoresis profiles with high resolution and reproducibility from immortalized human endocervical cell(H8) and cervical cancer cell(CaSKi),and to identify the differential expressions of cytoplasmic proteins.Methods:H8 cells and CaSKi cells were incubated,the cytoplasmic proteins of H8 cells and CaSKi cells were extracted and seperated by means of immobilized pH gradient-based two-dimensional gel electrophoresis(2-DE).The differential expression proteins were analyzed by using ImageMaster 2D analysis software.Results:We obtained well-resolved,reproducible 2-DE patterns,and 8 notable differential protein spots were defined in 2-DE gels.Conclusion:Cytoplasmic proteins of cervical precancerous lesion and cervical cancer are separated by means of immobilized pH gradient-based two-dimensional gel electrophoresis(2-DE).The expression maps of proteomics in cytoplasmic proteins of H8 and CaSKi are established.There are significant differences in proteome between cytoplasmic proteins of H8 cells and CaSKi cells.These data may be valuable for differential proteomics research of cervical precancerous lesion and cervical cancer.