Progress of the relationship between S100A9 and breast cancer / 肿瘤研究与临床

S100A9 is a calcium-binding protein that plays an important role in the progression of malignant tumors. Related studies have confirmed that the abnormal expression of S100A9 is closely related to the proliferation, metastasis and prognosis of breast cancer, while whether S100A9 can be used as a marker for the diagnosis and treatment of breast cancer is still controversial. This article reviews the current research status of S100A9 and its application prospect in the development, progression, diagnosis and treatment of breast cancer.

MANF brakes TLR4 signaling by competitively binding S100A8 with S100A9 to regulate macrophage phenotypes in hepatic fibrosis

The mesencephalic astrocyte-derived neurotrophic factor (MANF) has been recently identified as a neurotrophic factor, but its role in hepatic fibrosis is unknown. Here, we found that MANF was upregulated in the fibrotic liver tissues of the patients with chronic liver diseases and of mice treated with CCl4. MANF deficiency in either hepatocytes or hepatic mono-macrophages, particularly in hepatic mono-macrophages, clearly exacerbated hepatic fibrosis. Myeloid-specific MANF knockout increased the population of hepatic Ly6Chigh macrophages and promoted HSCs activation. Furthermore, MANF-sufficient macrophages (from WT mice) transfusion ameliorated CCl4-induced hepatic fibrosis in myeloid cells-specific MANF knockout (MKO) mice. Mechanistically, MANF interacted with S100A8 to competitively block S100A8/A9 heterodimer formation and inhibited S100A8/A9-mediated TLR4-NF-κB signal activation. Pharmacologically, systemic administration of recombinant human MANF significantly alleviated CCl4-induced hepatic fibrosis in both WT and hepatocytes-specific MANF knockout (HKO) mice. This study reveals a mechanism by which MANF targets S100A8/A9-TLR4 as a "brake" on the upstream of NF-κB pathway, which exerts an impact on macrophage differentiation and shed light on hepatic fibrosis treatment.

Predictive value of S100A9 for lymph node metastasis in cervical cancer / 中南大学学报(医学版)

OBJECTIVES@#Lymph node metastasis affects the initial treatment strategy for cervical cancer and is hard to be diagnosed in clinical practice.This paper aims to explore the relationship between calcium-binding A9 (S100A9) and lymph node metastasis (LNM) in cervical cancer, and to determine the predictive value of S100A9 for LNM in cervical cancer.@*METHODS@#We performed a retrospective cohort study and collected the pathological data, follow-up data, and paraffin tissue samples of 99 patients with cervical cancer who underwent modified extensive or extensive hysterectomy plus pelvic lymphadenectomy at the Department of Gynecology, Xiangya Hospital, Central South University from January 2013 to December 2018. Immunohistochemistry was used to detect the expression of S100A9 in cervical cancer tissues, and the correlation between S100A9 expression and LNM of cervical cancer, or clinicopathological characteristics were analyzed. The receiver operating characteristic (ROC) curve was used to establish a predictive model for LNM of cervical cancer, and Chi-square test of four-grid table was used to evaluate the diagnostic value of S100A9 for LNM in cervical cancer.@*RESULTS@#The expression of S100A9 was significantly correlated with LNM. The S100A9 immunohistochemical semi-quantitative score of the LNM group was significantly higher than that in the non-lymph node metastasis group (<0.001). Moreover, the expression of S100A9 was significantly correlated with histological type, stromal invasion, lymphatic vessel invasion, or LNM (<0.05). The cut-off of the ROC curve for predicting LNM was 5, with the Youden index of 0.649 and the area under the ROC curve of 0.863. The disease-free survival and overall survival in the S100A9 positive group were significantly shorter than those in the negative group (<0.05). S100A9 alone had a sensitivity of 71.4%, a specificity of 91.5%, and an accuracy of 85.1% for diagnosing LNM. Imaging had a sensitivity of 32.1%, a specificity of 74.6%, and an accuracy of 60.9%. Combination of S100A9 with image examination in parallel test had a sensitivity of 85.7%, a specificity of 71.2%, and an accuracy of 75.9%, while combination of S100A9 and image examination in serial test had a sensitivity of 17.9%, a specificity of 98.3%, and an accuracy of 72.4%.@*CONCLUSIONS@#S100A9 may be associated with LNM in cervical cancer. S100A9 shows a promising perspective in predicting LNM in cervical cancer. Combination of S100A9 and image examination in serial test has a high specificity for LNM.

Expression of S100A8 and S100A9 in oviduct epithelial cells induced by estrogen and their possible role in oviduct / 中国药理学通报

Aim To study whether the genes S100A8 and S100A9 are related to the functional regulation of oviduct by estrogen, and to explore their possible effects on fallopian tubes. Methods The basic expression and distribution of S100A8 and S100A9 in ampullary oviduct tissues of healthy sheep in diestrum were verified by immunohistochemistry. The expressions of S100A8 and S100A9 (mRNA and protein) in oviduct epithelial cells were detected by q-PCR and immunofluorescence, the cells were treated by E2 at different time points and different concentrations. Results S100A8 and S100A9 were highly expressed in mucosal epithelium and glandular epithelium of sheep uterine tubes during the diestrum period, and in blood vessels as well. The expression of S100A8 and S100A9 in tubal epithelial cells changed dynamically at different time ponits under the action of high concentration of E2 in vitro, and reached the peak 6 hours after E2 treatment. At this time, different concentrations of E2 significantly induced the high expression of S100A8 and S100A9, but the expressions of S100A8 and S100A9 were the highest at the concentrations of 10-7 mol • L-1 and 10-8 mol • L-1 , and there was no significant difference between the two groups. Conclusions S100A8 and S100A9 in oviduct epithelial cells are regulated by estrogen. Under the regulation of high concentration estrogen, the high expression of SI00A8 and S100A9 may be related to the natural defense of reproductive tract during mating, and may be involved in the transport of eggs in fallopian tube.

Construction and anti-tumor efficacy of a peptide vaccine that targets calcium-binding protein S100A9 / 药学学报

Calcium-binding protein S100A9 is closely related to inflammation and tumor invasion, and is one of the specific markers of myeloid-derived suppressor cells (MDSC). In this study, a recombinant polypeptide vaccine CTB-S100A9 targeting mouse calcium-binding protein S100A9 was constructed by fusion cholera toxin B subunit (CTB) with S100A9 gene. The CTB-S100A9 fusion protein was expressed in E coli. and purified by Ni+ affinity chromatography. Vaccinate the purified recombinant CTB-S100A9 protein supplemented with aluminum hydroxide adjuvant can break the autoimmune tolerance and produce high titer of S100A9 antibody in mice. Moreover, the S100A9 antibody produced by CTB-S100A9 vaccination is more specific and does not cross-react with S100A8. In the mouse 4T1 breast cancer model, CTB-S100A9 vaccination not only has significant tumor prevention effects, but also has significant tumor therapeutic effects. In addition, CTB-S100A9 significantly inhibited lung metastasis in 4T1 mice breast cancer model. Further analysis by flow cytometry showed that CTB-S100A9 vaccination can significantly reduce the tumor induced Treg cells and granulocyte-derived MDSC in 4T1 mice model, and reverse the tumor immunosuppressive environment, thereby promote the anti-tumor efficacy. The animal experiments in this study were carried out under the animal care guidelines approved by the Animal Ethics Committee of the Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine. This study shows that CTB-S100A9 is a good recombinant vaccine that targets the tumor immune-suppression environment and has great potential for the future clinical application.

Prognostic Role of S100A8 and S100A9 Protein Expressions in Non-small Cell Carcinoma of the Lung

BACKGROUND: S100A8 and S100A9 have been gaining recognition for modulating tumor growthand metastasis. This study aimed at evaluating the clinical significance of S100A8 and S100A9 innon-small cell lung cancer (NSCLC). METHODS: We analyzed the relationship between S100A8and S100A9 expressions, clinicopathological characteristics, and prognostic significance in tumorcells and peritumoral inflammatory cells. RESULTS: The positive staining of S100A8 in tumorcells was significantly increased in male (p < .001), smoker (p = .034), surgical method other thanlobectomy (p = .024), squamous cell carcinoma (SQCC) (p < .001) and higher TNM stage (p = .022)compared with female, non-smoker, lobectomy, adenocarcinoma (ADC), and lower stage. Theproportion of tumor cells stained for S100A8 was related to histologic type (p < .001) and patientsex (p = .027). The proportion of inflammatory cells stained for S100A8 was correlated with patientage (p = .022), whereas the proportion of inflammatory cells stained for S100A9 was correlatedwith patient sex (p < .001) and smoking history (p = .031). Moreover, positive staining in tumorcells, more than 50% of the tumor cells stained and less than 30% of the inflammatory cellsstained for S100A8 and S100A9 suggested a tendency towards increased survivability in SQCCbut towards decreased survivability in ADC. CONCLUSIONS: S100A8 and S100A9 expressions might be potential prognostic markers in patients with NSCLC.

Study on the proliferation and the expression of S100A8 and S100A9 in Helicobacter pylor-infected human gastric epithelial cells GES-1 / 中华消化杂志

Objective@#To investigate the effects of Helicobacter pylori (H.pylori) on the proliferation of GES-1 cells and the expressions of S100A8 and S100A9 in human gastric epithelial cell line GES-1. @*Methods@#H. pylori were co-cultured with GES-1 cells at different infection plural (muhiplieity of infection (MOI) 50∶1, 100∶1, 200∶1), then the cells and cell culture supernatants were collected. The proliferative activity was detected by cell counting kit 8 (CCK-8) methods. The expression of S100A8 and S100A9 at mRNA level was determined by reverse transcription polymerase chain reaction (RT-PCR). The levels of S100A8 and S100A9 proteins and tumor necrosis factor-alpha (TNF-α) in cell culture supernatants were measured by enzyme linked immunosorbent assay (ELISA). T test and Pearson method were performed for statistical analysis. @*Results@#The negative control group was taken as the baseline, the proliferation rates of GES-1 cells of the H. pylori multiplicity 50∶1 group, 100∶1 group and 200∶1 group were (105.51±4.78)%, (168.97±11.29)% and (64.05±10.11)%, respectively. There was no statistically significant difference in the proliferation rate of GES-1 cells between H. pylori multiplicity 50∶1 group and negative control group (t=0.69, P=0.51). The proliferation rate of GES-1 cells of the H. pylori multiplicity 100∶1 group was higher than that of the negative control group, and the difference was statistically significant (t=10.63, P<0.01). The proliferation rate of GES-1 in the H. pylori multiplicity 200∶1 group was lower than that of the negative control group, and the difference was statistically significant (t=-5.54, P<0.01). The expression of S100A8 and S100A9 at the mRNA level in the H. pylori multiplicity 200∶1 group was 0.31±0.21 and 8.66±4.08, respectively, which were higher than those of the negative control group (0.06±0.05 and 0.08±0.08), and the differences were statistically significant (t=10.20 and 6.89, both P<0.05). The expressions of S100A8 at the protein level of H. pylori multiplicity 50∶1, 100∶1, and 200∶1 groups were (112.21±1.25) ng/mL, (120.39±1.61) ng/mL and (121.28±0.71) ng/mL, respectively; while the expression of S100A9 at the protein level were (179.43±2.44) ng/mL, (191.47±1.98) ng/mL and (201.80±2.06) ng/mL, respectively; and the expression of TNF-α levels were (285.52±3.64) ng/mL, (320.08±2.28) ng/mL and (350.97±2.90) ng/mL, respectively; which were all higher than those of the negative control group ((76.14±1.30) ng/mL, (161.35±1.31) ng/mL and (270.08±2.96) ng/mL, respectively), and the differences were statistically significant (tS100A8=35.09, 43.06, 43.92, tS100A9 = 11.13, 18.54, 24.90, tTNF-α= 6.34, 20.54, 33.23; all P<0.01). The expressions of S100A8 and S100A9 at the protein level were positively correlated with TNF-α (r=0.92 and 0.95, both P<0.01). @*Conclusion@#S100A8 and S100A9 may be involved in the process of H. pylori induced proliferation disorder and inflammation in GES-1 cells.

Extremely elevated IL-18 levels may help distinguish systemic-onset juvenile idiopathic arthritis from other febrile diseases

The aim of this research was to explore whether IL-18 can be a serological marker for the diagnosis of systemic-onset juvenile idiopathic arthritis (sJIA). A total of 23 sJIA patients (13 males, median age 8.2), 20 acute lymphoblastic leukemia (ALL) patients, 18 patients with severe infections (SIF), 26 Kawasaki disease (KD) patients, 18 juvenile idiopathic arthritis (JIA) patients, and 25 healthy control patients were selected for this study. Enzyme-linked immunosorbent assays (ELISAs) were used to determine the serum concentrations of the S100A8, S100A9, and IL-6 proteins. The serum IL-18 levels were detected by a cytometric bead array (CBA). The serum IL-6 concentrations in various disease groups were significantly higher than that in the healthy control group. The IL-6 concentrations exhibited no significant difference between disease groups. The S100A8 level in the sJIA group was significantly higher than those of the ALL, JIA, and healthy control groups but showed no significant difference compared to the SIF and KD groups. The S100A9 serum concentration in the sJIA group was significantly higher than those in the ALL and healthy control groups and exhibited no significant difference from the SIF, KD, and JIA groups. The IL-18 level of the sJIA group was significantly higher than that of the other febrile disease groups. The IL-18 serum concentration may be used as a biological serum marker to distinguish sJIA from other febrile diseases.

Clinical significance of serum calcium-binding protein A9(S100A9)levels in patients with rheumatoid ar- thritis / 实用医学杂志

Objective To investigate the serum level of calcium-binding protein A9(S100A9)in patients with rheumatoid arthritis and explore its potential clinical significance. Methods The serum level of S100A9 was measured by ELISA in 79 rheumatoid arthritis (RA) patients and 20 healthy controls (HC). The correlation of S100A9 level and relevant clinical parameters were analyzed. Results The serum level of S100A9 was significantly increase in RA patients than those in HC. The serum level of S100A9 was higher in high disease activity than that in moderate and low disease activity in the RA group. There was a positive correlation in serum S100A9 level and DAS28 score,Rheumatoid factor,tender joint count and swollen joint count. Conclusion The serum level of S100A9 increases in RA patients and correlates with RA activity.

Effects of denatonium benzoate on calcium binding protein S100A9 and mucus in the airways of asthmatic mice / 医学研究生学报

number of goblet cells, mucus secretion and mucin MUC5AC content in lung tissues. Results S100A9 in BALF of group B was (11.89±0.77) ng/mL, S100A9 integrated optical density (IOD) value in airway epithelial cells was 13.96±1.62, PAS stain area /epithelial cell area was (12.53±1.21)%, relative value of MUC5AC / NADPH was 173.91±4.29, all of the above were higher than those of group A [(6.19±0.61) ng/mL, 4.97±0.30, (1.94±0.18)%, 1];S100A9 levels, IOD of S100A9 in airway epithelial cells, PAS stain area / epithelial cell area (%), relative value of MUC5AC / NADPH in group C [(10.69±0.79) ng / ml, 11.80±0.72, (10.61±0.61)%, 94.65±1.59], group D[(9.49±0.99) ng/mL, 10.39±0.59, (8.63±0.62)%, 82.08±1.12], group E [(7.54± 0.42) ng/mL, 5.63±0.84, (4.59±0.87)%, 26.30±1.94] were lower than group B, which showed a dose-dependent reduction and the difference was statistically significant (P<0.05 or P<0.01). Conclusion DB downregulates the expression level of Ca2+-binding protein S100A9 and the mucus secretion amount of the airway goblet cells in rats.

Expression and effect of S100A9 in the periodontium of diabetic rats / 华西口腔医学杂志

<p><b>OBJECTIVE</b>The study seeks to investigate the expression of S100A9 and its potential role in periodontal diseases induced by diabetes.</p><p><b>METHODS</b>A diabetic SD rat model was established through intraperitoneal injection of streptozotocin (STZ). Hematoxylin-eosin (HE) staining was performed to study the structure of the periodontium of diabetic rats. Using immunohistochemical staining, the distribution of S100A9 expression was detected in the periodontium of diabetic rats. Ex-pressions of Toll-like receptor 4 (TLR4) (ligands of S100A9) and p-P65/nuclear factor κB (NF-κB) were also measured.</p><p><b>RESULTS</b>The trabecular structure of alveolar bone was sparser, and lamina dura was disappeared in diabetic rats. Obviously higher expressions of S100A9 were observed in the periodontal ligament, alveolar bone, and gingival epithelial of diabetic rats than in the control rats. TLR4 expressions in the periodontal ligament, alveolar bone and gingival epithelial of the diabetic rats were also higher as compared to the control rats. p-P65 expression was not detected in the control rats, but was detected in the periodontal ligament and alveolar bone of the diabetic rats.</p><p><b>CONCLUSIONS</b>Periodontium lesions in diabetes mellitus may be induced by the activation of TLR4 and NF-κB signaling pathway meditated by S100A9. 
.</p>

Development and identification of mouse anti-human S100 A9 natural protein / 中国免疫学杂志

Objective:To prepare and identify the mouse anti-human monoclonal antibodies ( mAbs) using leukocytes as im-munogens. Methods: The mice were immunized using human peripheral blood leukocytes. Then, use of B lymphocyte hybridoma technology preparation of mAbs,followed screening by immunocytochemistry and limited dilution. The secreted mAbs were identified by immunoprecipitation,mass spectrometry,Western blot,ELISA and immunohistochemistry. Results:The 35 positive polyclonal cells were obtained,of which 11 strains secreted mAbs against S100A9. And one strain was used to prepare monoclonal antibody. The purified mAb against S100A9 were purified and identified as IgG1 subtype,with the titer,purity and affinity constant was 1∶3. 18×105,95% and 3. 54×108 L/mol,respectively. This mAb generally had 0. 12% crossed reactivity to S100A8 ,and showed little or no cross reactivity to S100A12 and S100A13. The prepared monoclonal antibodies can specifically recognizes the S100A9 antigen in human breast cancer tissues. Conclusion:Successful preparation of mAb against S100A9,which can secrete specific mAb against S100A9 protein with high titers and specificity have been established successfully,which laid the foundation for the immunology application.

Study on the mechanism of S100A9-induced secretion of vascular endothelial growth factor-A by monocytes / 国际检验医学杂志

Objective To explore the molecular mechanism of S100A9-induced secretion of vascular endothelial growth factor-A (VEGF-A)by monocytes.Methods Peripheral blood specimen were collected from healthy individuals undergoing physical exami-nation and the CD14 + monocytes were purified by using immunomagnetic beads and the expression of the receptor for advanced gly-cation endproducts (RAGE)was detected by flow cytomertry.In vitro CD14 + monocytes were stimulated by S100A9,and anti-RAGE antibody or NK-κB signal pathway inhibitor were pre-incubated for 1 hour and then stimulated by S100A9,the levels of VEGF-A were detected by using enzyme-linked immunosorbent assay.Results The high level of RAGE was expressed by isolated CD14 + monocytes,after S100A9 stimulation,the secretion of VEGF-A by CD14 + monocytes was significantly increased in a dose and time dependent manner.However,the inducing VEGF-A was significantly decreased(P <0.01 ),while pre-treated with anti-RAGE antibody or NK-κB inhibitor (P <0.01).Conclusion S100A9 inducing the secretion of VEGF-A by monocytes and is de-pended on RAGE-NK-κB signal pathway,suggesting that S100A9 might promote angiogenesis.

Urinary Nucleic Acid TSPAN13-to-S100A9 Ratio as a Diagnostic Marker in Prostate Cancer

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.

Effect of spleen lymphocytes on the splenomegaly in hepatocellular carcinoma-bearing mice / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study the effect of spleen lymphocytes on the splenomegaly by hepatocellular carcinoma-bearing mouse model.</p><p><b>METHODS</b>Cell counts, cell cycle distribution, the percentage of lymphocytes subsets and the levels of IL-2 were measured, and two-dimensional gel electrophoresis (2-DE) was used to investigate the relationship between spleen lymphocytes and splenomegaly in hepatocellular carcinoma-bearing mice.</p><p><b>RESULTS</b>Compared with the normal group, the thymus was obviously atrophied and the spleen was significantly enlarged in the tumor-bearing group. Correlation study showed that the number of whole spleen cells was positively correlated with the splenic index. The cell diameter and cell-cycle phase distribution of splenocytes in the tumor-bearing group showed no significant difference compared to the normal group. The percentage of CD3+ T lymphocytes and CD8+ T lymphocytes in spleen and peripheral blood of tumor-bearing mice were substantially higher than that in the normal mice. Meanwhile, the IL-2 level was also higher in the tumor-bearing group than in the normal group. Furthermore, two dysregulated protein, β-actin and S100-A9 were identified in spleen lymphocytes from H22-bearing mice, which were closely related to cellular motility.</p><p><b>CONCLUSION</b>It is suggested that dysregulated β-actin and S100-A9 can result in recirculating T lymphocytes trapped in the spleen, which may explain the underlying cause of splenomegaly in H22-bearing mice.</p>

S100A8/A9 as a biomarker for synovial inflammation and joint damage in patients with rheumatoid arthritis

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.

The Expression of Interleukin-22 and S100A7, A8, A9 mRNA in Patients with Psoriasis Vulgaris / 华中科技大学学报（医学）（英德文版）

In order to study the expression of intedeukin-22 (IL-22) and S100A7, A8, A9 mRNA in the skin lesions of patients with psoriasis vulgaris and their relationship, the biopsies were taken from skin lesions in 35 patients with psoriasis vulgaris and the skin of 16 normal controls, and the expres- sion levels of IL-22 and S 100A7, A8 and A9 mRNA were detected by semi-quantitative RT-PCR. The results showed that (1) IL-22 and SI00A8, A9 mRNA were positively expressed in the psoriatic skin lesions but negatively expressed in the normal controls; The expression level of S100A7 was (1.133±0.040) in the psoriatic skin lesions, significantly higher than that in the normal controls (0.744±0.037, P<0.01). (2) There were significantly positive correlations between the expression of IL-22/S100A7 mRNA, IL-22/S100A8 mRNA, IL-22/S100A9 mRNA in the psoriasis vulgaris (r1=0.543, r2=0.774, r3=0.621, P<0.01). It was concluded that IL-22 and S100A7, A8, A9 might play important roles in the occurrence and progression of psoriasis.