S100A8 and S100A9 are major leukocyte proteins, known as damage-associated molecular patterns, found at high concentrations in the synovial fluid of patients with rheumatoid arthritis (RA). A heterodimeric complex of S100A8/A9 is secreted by activated leukocytes and binds to Toll-like receptor 4, which mediates downstream signaling and promotes inflammation and autoimmunity. Serum and synovial fluid levels of S100A8/A9 are markedly higher in patients with RA than in patients with osteoarthritis or miscellaneous inflammatory arthritis. Serum levels of S100A8/A9 are significantly correlated with clinical and laboratory markers of inflammation, such as C-reactive protein, erythrocyte sedimentation rate, rheumatoid factor, and the Disease Activity Score for 28 joints. Significant correlations have also been found between S100A8/A9 and radiographic and clinical assessments of joint damage, such as hand radiographs and the Rheumatoid Arthritis Articular Damage score. In addition, among known inflammatory markers, S100A8/A9 has the strongest correlation with total sum scores of ultrasonography assessment. Furthermore, baseline levels of S100A8/A9 are independently associated with progression of joint destruction in longitudinal studies and are responsive to change during conventional and biologic treatments. These findings suggest S100A8/A9 to be a valuable diagnostic and prognostic biomarker for RA.
Arthritis, Rheumatoid/*blood/pathology/radiography ; Arthrography ; Biological Markers/blood ; Calgranulin A/*blood ; Calgranulin B/*blood ; Humans ; Joints/pathology ; Synovial Fluid/metabolism

<b>OBJECTIVEb>To investigate the mRNA and protein expression levels of S100A8 and S100A9 in giant cell tumor (GCT) of bone, and its relation with radiological findings and biological behavior.

<b>METHODSb>Forty three patient with GCT of bone admitted in Ruijin Hospital Shanghai Jiaotong University School of Medicine from January 2009 to June 2012 were enrolled in the study. The expression levels of S100A8 and S100A9 mRNA and protein were detected by using semiquantitative RT-PCR and Western blotting in 43 specimens of GCT and 6 specimens of normal bone marrow. The CT and MRI findings of patients were retrospectively reviewed, its relation with tissue expression of S100A8 and S100A9 was analyzed.

<b>RESULTSb>Among 43 GCT cases 40 showed positive expression of S100A8 and S100A9 mRNA and protein, and the expression levels were significantly higher than those in normal bone marrow P<0.05). The expression level of S100A8 protein was significantly different in bone GCT with different composition ratio on MRI (P<0.05).The expression level of S100A9 protein was significantly different in GCT with different degree of bone destruction on CT scan (P<0.05).

<b>CONCLUSIONb>The expression of S100A8 and S100A9 mRNA and protein is up-regulated in GCT of bone. The expression of S100A8 and S100A9 is associated with the real composition ratio and the degree of bone destruction, respectively, indicating that S100A8 and S100A9 may be involved in the biological behavior of bone GCT.

Bone Neoplasms ; metabolism ; Calgranulin A ; metabolism ; Calgranulin B ; metabolism ; China ; Giant Cell Tumor of Bone ; metabolism ; Humans ; RNA, Messenger ; Tomography, X-Ray Computed ; Up-Regulation

<b>BACKGROUNDb>S100A8 and S100A9 are two members of the S100 protein family characterized by the presence of two Ca2+-binding sites of the EF-hand type. Previous studies suggested that the whole S100 family displays significant functions in tumor growth, progression and invasion. This study aimed to determine the expression of the two indices of the family, S100A8 and S100A9, in lung cancer tissues and normal lung tissues and its correlation with clinical features.

<b>METHODSb>A total of 60 cases with a variety of clinical data that were diagnosed with different histological subtypes of lung cancer were investigated. Semi-quantitative reverse transcriptase-PCR (Sq-Rt-PCR) and immunohistochemical staining of cancer, adjacent and peripheral lung tissues were executed to distinguish the expression patterns of S100A8 and S100A9 and to further clarify their correlation with clinical features.

<b>RESULTSb>Immunohistochemical staining of both proteins showed a significant up-regulation in lung cancer tissue (S100A8, S100A9, P<0.0001), and PCR revealed that the levels of S100A8 and S100A9 expression were significantly higher in lung cancer tissues (S100A8 P=0.002/0.004; S100A9 P=0.022/0.026). The higher expression was found to be correlated with the clinical characteristics of adenocarcinoma, inflammation and stage IV lesion.

<b>CONCLUSIONSb>S100A8, S100A9 up-regulation was found in the lung adenocarcinoma and end stage lung cancer tissue, the correlation of which with their higher expression in inflammatory lung tissues may indicate the collaborative effect of inflammation on the progression of cancer.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Aged ; Calgranulin A ; genetics ; metabolism ; Calgranulin B ; genetics ; metabolism ; Female ; Humans ; Immunohistochemistry ; Inflammation ; genetics ; metabolism ; pathology ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Reverse Transcriptase Polymerase Chain Reaction

<b>OBJECTIVEb>To explore mechanism of S100A8 in the oncogenesis and development of laryngeal cancer.

<b>METHODSb>Proteins interacting with S100A8 were isolated from laryngeal cancer cell lines Hep-2 by immunoprecipitation assay with anti-S100A8 antibody. The target bands were cut out and identified by maxtrix assisted laser desorption/ionization time of flight (MALDI-TOF). The peptide mass fingerprinting data of the proteins identified were analyzed based on the Mascot database. The NF-kappa B binding sites of the proteins were predicted by P-Match software. The binding ability of one of the proteins to S100A8 was confirmed by co-immunoprecipitation and immunocytochemistry methods.

<b>RESULTSb>Four proteins interacting with S100A8 were obtained, which were hypothetical protein LOC80154, MHC class I HLA-B, similar to T-box 1 isoform C and sarcolemmal associated protein 1. The four genes were predicted to have NF-kappa B binding sites. MHC class I HLA-B, which is one of targets in NF-kappa B pathway, was first confirmed to have the binding ability to S100A8.

<b>CONCLUSIONb>The novel partners of S100A8 identified in the study might be involved in NF-kappa B pathway. The binding ability of MHC class I HLA-B to S100A8 implies that S100A8 might function as a new member with other proteins including HLA-B in NF-kappa B pathway. These findings provide a new clue to further study on the molecular mechanism of S100A8 in the genesis of laryngeal carcinomas.

Animals ; Binding Sites ; Calgranulin A ; genetics ; metabolism ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; HLA-B Antigens ; genetics ; metabolism ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; pathology ; NF-kappa B ; metabolism ; Signal Transduction

OBJECTIVE: To observe the correlation between the expression of galectin-7 and S100A9 with the development of cervical squamous carcinoma. METHODS: Immunohistochemical SP staining was used to detect the expression of galectin-7 and S100A9 in 243 patients with cervical intraepithelial neoplasia (CIN) or cervical squamous carcinoma. The association of clinical data with galectin-7 and S100A9 expression was examined. RESULTS: The expression of galectin-7 and S100A9 in CIN and cervical squamous carcinoma was significantly different (P<0.05). The positive rates of galectin-7 in normal cervical tissues, CIN I, CIN II, CIN III, and cervical squamous carcinoma were 56.7%, 41.9%, 32.0%, 27.3%, and 25.0%, respectively. Statistic analysis found significant difference between the normal cervical tissues and cervical squamous carcinoma (P<0.0045). The positive rates of S100A9 in CIN I, CIN II, CIN III, and cervical squamous carcinoma were 80.0%, 77.4%, 48.0%, 27.3%, and 20.2%. Statistic analysis showed significant difference between the normal tissues and CIN III, the normal cervical tissues and cervical squamous carcinoma, CIN I and CIN III, CIN I and cervical squamous carcinoma, CIN II and cervical squamous carcinoma (P<0.0045). A positive correlation was found between galectin-7 and S100A9 expression in CIN and cervical squamous carcinoma (rs=0.298, P<0.001). Expressions of both galectin-7 and S100A9 in cervical squamous carcinoma were associated with the clinical stage and lymph nodes (P<0.05), but not with patient's age and degree of differentiation (P>0.05). Expression of galectin-7 was associated with the survival rate of patients with cervical squamous carcinoma (P<0.05). Univariate analysis of Cox proportional hazards regression model revealed that the FIGO stage, lymph nodes metastasis, and the expression of galectin-7 were relevant to the 5 year survival rate of patients with cervical squamous carcinoma, which was confirmed by multiple analysis of Cox proportional hazards regression model. CONCLUSION Expression of galectin-7 and S100A9 is related with cervical the tumorigenesis of carcinoma, clinical stage, and lymph nodes of cervical squamous carcinoma. Galectin-7 is probably associated with the prognosis. The long-term survival of patients with cervical carcinoma may be associated with FIGO stage, lymph node metastasis, and the expression of galectin-7.
Calgranulin B ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; Cell Differentiation ; Cervical Intraepithelial Neoplasia ; metabolism ; Female ; Galectins ; metabolism ; Humans ; Lymphatic Metastasis ; Prognosis ; Proportional Hazards Models ; Survival Rate ; Uterine Cervical Neoplasms ; metabolism

<b>OBJECTIVEb>To explore the differentially expressed proteins in normal cervix, cervical intraepithelial neoplasia (CIN) and squamous cervical carcinoma (SCC) tissues by differential proteomics, and to provide a basis for studies on CIN molecular pathogenesis, clinical diagnosis and treatment.

<b>METHODSb>Uterine cervical tissue specimens from the patients treated between August 2008 and September 2009 in the Department of Oncology of Beijing Obstetrics and Gynecology Hospital were collected. There were samples of normal cervix (n = 9), CIN (n = 23, CIN I = 7, CIN II = 8, CIN III = 8) and SCC (n = 7). 2-D DIGE and DeCyder software were used to detect the differentially expressed protein-spots. Then MALDI-TOF/TOF MS was used to analyze the differentially expressed proteins. Collect normal cervix(n = 20), CIN (n = 60) and SCC (n = 20), immunohistochemistry (IHC) and Western blot were used to verify the differentially expressed proteins of S100A9 (S100 calcium-binding protein A9) , eEF1A1 (eukaryotic elongation factor 1-alpha-1) and PKM2 (pyruvate kinase isozymes M2) among the normal cervix, CIN and SCC tissues. Immunohistochemistry was used to detect the differentially expressed S100A9, eEF1A1 and PKM2 in the cervical tissues.

<b>RESULTSb>2D gel electrophoresis images with high resolution and good repeatability were obtained. Forty-six differentially expressed proteins (27 were up-regulated and 19 were down-regulated) were selected among the normal, CIN, and SCC, and 26 proteins were successfully identified. Immunohistochemistry showed that protein S100A9 was mainly expressed in the cytoplasm, and its positive expression rate was 20.0% in normal cervical mucosa, 70.0% in CIN, and 100.0% in squamous cell carcinoma, with a significant difference between them (P = 0.006). eEF1A1 was mainly expressed in the cell plasma. Its positive expression rate was 70.0% in normal cervix, 73.3%in CIN and 60.0% in SCC tissues, with a non-significant difference between them (P = 0.758). The protein PKM2 was mainly expressed in the cell nuclei. Its positive expression rate was 100.0% in normal cervix, 93.3% in CIN and 75.0% in SCC tissues, showing a difference close to statistical significance (P = 0.059) between them. The results of Western blot were similar with that of immunohistochemical examination.

<b>CONCLUSIONSb>There are differentially expressed proteins among normal cervix, CIN and SCC. S100A9, eEF1A1 and PKM2 may become candidate markers for early diagnosis of cervical cancer and new targets for therapy. It also provides a further basis for studies of the pathogenetic mechanism of CIN developing to cervical cancer.

Adult ; Aged ; Biomarkers, Tumor ; metabolism ; Calgranulin B ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; Carrier Proteins ; metabolism ; Cervical Intraepithelial Neoplasia ; metabolism ; Cervix Uteri ; metabolism ; Female ; Humans ; Immunohistochemistry ; Membrane Proteins ; metabolism ; Middle Aged ; Peptide Elongation Factor 1 ; metabolism ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Thyroid Hormones ; metabolism ; Uterine Cervical Neoplasms ; metabolism ; Young Adult

Myeloid-related protein (MRP)8/MRP14 is an endogenous Toll-like receptor 4 (TLR4) ligand and is abundant in synovial fluid (SF) of rheumatoid arthritis (RA) patients. Belonging to damage-associated molecular patterns, it amplifies proinflammatory mediators and facilitates a wide range of inflammatory and autoimmune diseases. Interleukin (IL)-17-producing T-helper (Th)17 cells have a crucial role in RA pathogenesis, and IL-6 is the key factor promoting Th17 differentiation. We investigated whether the level of MRP8/MRP14 is positively associated with IL-6 and IL-17 levels in RA SF and found that MRP8/MRP14 level had a significant correlation with IL-6 and IL-17 levels in RA SF. We also observed that MRP8-induced IL-17 production by peripheral blood mononuclear cells but MRP14 did not. Upon stimulation with MRP8, IL-6 production was enhanced by RA fibroblast-like synoviocytes (FLS) and was further elevated by coculturing RA FLS with activated CD4+ T cells. Moreover, we demonstrated that MRP8-activated IL-6 production by RA FLS promoted differentiation of Th17 cells using the coculture system consisting of CD4+ T cells and RA FLS. In addition, IL-6 blockade attenuated Th17 polarization of CD4+ T cells in the cocultures. Inhibitor studies revealed that MRP8 increased IL-6 production in RA FLS via TLR4/phosphoinositide 3-kinase/nuclear factor-kappaB and mitogen-activated protein kinase signaling pathways. Our results show that MRP8 has a crucial role in stimulating IL-6 expression by RA FLS, and subsequently promotes Th17 differentiation in RA, suggesting that neutralizing MRP8 level in RA synovium may be an effective therapeutic strategy in RA treatment.
ATP-Binding Cassette Transporters/*metabolism ; Adult ; Aged ; Arthritis, Rheumatoid/*pathology ; CD4-Positive T-Lymphocytes/metabolism ; Calgranulin B/metabolism ; Cell Differentiation/*immunology ; Fibroblasts/*metabolism/pathology ; Humans ; Inflammation Mediators/metabolism ; Interleukin-17/metabolism ; Interleukin-6/*biosynthesis ; Middle Aged ; Signal Transduction/immunology ; Synovial Fluid/cytology ; Synovial Membrane/metabolism/pathology ; Th17 Cells/*pathology ; Toll-Like Receptor 4/metabolism ; *Up-Regulation

The potential use of urinary nucleic acids as diagnostic markers in prostate cancer (PCa) was evaluated. Ninety-five urine samples and 234 prostate tissue samples from patients with PCa and benign prostatic hyperplasia (BPH) were analyzed. Micro-array analysis was used to identify candidate genes, which were verified by the two-gene expression ratio and validated in tissue mRNA and urinary nucleic acid cohorts. Real-time quantitative polymerase chain reaction (qPCR) was used to measure urinary nucleic acid levels and tissue mRNA expression. The TSPAN13-to-S100A9 ratio was selected to determine the diagnostic value of urinary nucleic acids in PCa (P = 0.037) and shown to be significantly higher in PCa than in BPH in the mRNA and nucleic acid cohort analyses (P < 0.001 and P = 0.013, respectively). Receiver operating characteristic (ROC) analysis showed that the area under the ROC curve was 0.898 and 0.676 in tissue mRNA cohort and urinary nucleic acid cohort, respectively. The TSPAN13-to-S100A9 ratio showed a strong potential as a diagnostic marker for PCa. The present results suggest that the analysis of urine supernatant can be used as a simple diagnostic method for PCa that can be adapted to the clinical setting in the future.
Aged ; Aged, 80 and over ; Biomarkers, Tumor/*genetics/*urine ; Calgranulin B/*genetics ; Cohort Studies ; Humans ; Male ; Middle Aged ; Nucleic Acids/*genetics/*urine ; Oligonucleotide Array Sequence Analysis ; Prostate/metabolism ; Prostatic Hyperplasia/diagnosis/genetics/urine ; Prostatic Neoplasms/diagnosis/*genetics/*urine ; RNA, Messenger/genetics/metabolism ; RNA, Neoplasm/genetics/metabolism ; ROC Curve ; Real-Time Polymerase Chain Reaction ; Tetraspanins/*genetics