Superexpressão do receptor de Inositol 1,4, 5-trifosfato isoforma 3 (ITPR3) em colangiocarcinoma e sua correlação com a proteína dependente de cálcio S100A4 / Inositol 1,4, 5-triphosphate receptor isoform 3 (ITPR3) overexpression in cholangiocarcinoma and its correlation with calcium-dependent protein S100A4

O colangiocarcinoma (CCA) é a segunda neoplasia mais maligna do fígado que surge na árvore biliar. O CCA está associado com mau prognóstico e os principais fatores envolvidos em sua patogênese não são bem compreendidos. Os receptores tirosina quinases (RTKs), como o receptor do fator de crescimento epidérmico (EGFR), podem mediar as vias de sinalização de cálcio intracelular (Ca 2+ ), via inositol 1,4,5-trifosfato (InsP3). Eles ativam os receptores 1,4,5-trifosfato (ITPRs) e regulam o crescimento tumoral. ITPR isoforma 3 é o principal canal de liberação intracelular de Ca 2+ em colangiócitos. Os efeitos do Ca 2+ intracelular, por sua vez são mediados por proteínas de ligação de cálcio, como calmodulina e proteína A4 de ligação de cálcio S100 (S100A4). No entanto, o significado clínico patológico e biológico de EGFR, ITPR3 e S100A4 no CCA permanece obscuro. Assim, o presente trabalho investiga a imuno exprepressão dessas três proteínas em 59 pacientes diagnosticados com CCA, submetidos a tratamento cirúrgico curativo e correlaciona os dados com características clínico-patológicas e sobrevida. A alta expressão de ITPR3 foi correlacionada com os níveis de CA 19-9, estágio TNM e metástases em linfonodos (N). Além disso, a expressão de ITPR3 foi aumentada em CCA distal em comparação com ductos biliares de controle e CCAs intra-hepáticos e peri-hilares. Os escores clínicos ITPR3 e S100A4 foram significativamente correlacionados. Em resumo, a super expressão de ITPR3 pode contribuir para a progressão da CCA e pode representar um potencial alvo terapêutico. Palavras-chave: ITPRs; ITPR3; S100A4; Colangiocarcinoma; Fígado; Câncer

Cholangiocarcinoma (CCA) is the second most malignant neoplasm in the liver that arises from the biliary tree. CCA is associated with a poor prognosis, and the key players involved in its pathogenesis are still not well understood. Receptor tyrosine kinases (RTKs), such as epidermal growth factor receptor (EGFR), can mediate intracellular calcium (Ca2+) signaling pathways via inositol 1,4,5trisphosphate (InsP3), activating inositol 1,4,5-trisphosphate receptors (ITPRs) and regulating tumor growth. ITPR isoform 3 (ITPR3) is the main intracellular Ca2+ release channel in cholangiocytes. The effects of intracellular Ca2+ are mediated by calciumbinding proteins such as Calmodulin and S100 calcium-binding protein A4 (S100A4). However, the clinicopathological and biological significance of EGFR, ITPR3 and S100A4 in CCA remains unclear. Thus, the present work investigates the immunoexpression of these three proteins in 59 CCAs from patients who underwent curative surgical treatment and correlates the data with clinicopathological features and survival. High ITPR3 expression was correlated with CA 19-9 levels, TNM stage and lymph node metastasis (N). Furthermore, ITPR3 expression was increased in distal CCA compared to control bile ducts and intrahepatic and perihilar CCAs. In summary, ITPR3 overexpression could contribute to CCA progression and it may represent a potential therapeutic target.

Immunolocalización de s100a4 y α-sma en tejidos gingivales de pacientes con hipertrofia gingival por tratamiento ortodóntico: estudio preliminar / Immunolocalization of s100a4 and α-sma in gingival tissues of patients with gingival hypertrophy due to orthodontic treatment: preliminary study

Objective: to determine the presence and distribution of markers of the epithelialmesenchymal transition (EMT) (S-100A4 and alpha-smooth muscle actin-α-SMA) in gingival tissues of patients affected by Gingival hypertrophy (GH) due to orthodontics.GH is an exaggerated increase in gingival tissue whose pathogenesis is unknown. However, it has been reported that the epithelial-mesenchymal transition as a process involved in other types of GH. Materials and methods: descriptive study that included the analysis of gingival tissues of healthy individuals (n = 6) and patients with GH by orthodontic treatment (n = 6). Before gingival surgery, the patients underwent a periodontal hygiene phase. The gingival tissue samples obtained were processed and embedded in paraffin. The cuts were made with a microtome and deposited on polysine adhesion slides. Histological hematoxylin-eosin staining was performed.The identification and location of S-100A4 and α-SMA markers was determined by immunohistochemistry with monoclonal antibodies. The reading of the findings was carried out by oral pathologists. Results: in healthy individuals, an S100A4 label was observed in Langerhans cells, while α-SMA was identified in the vascular endothelium of all samples analysed. However, in patients with GH due to orthodontics, they registered an intense staining of S100A4 in gingival fibroblasts, Langerhans cells, vascular endothelium, and areas adjacent to the rupture of blood vessel. α-SMA expression in GO was detected in the vascular endothelium and gingival fibroblasts. Conclusion: the differential immunostaining of EMT markers in gingival tissues of patients with orthodontic GH suggests an eventual role of EMT in the pathogenesis of this pathology..Au

Objective: to determine the presence and distribution of markers of the epithelialmesenchymal transition (EMT) (S-100A4 and alpha-smooth muscle actin-α-SMA) in gingival tissues of patients affected by Gingival hypertrophy (GH) due to orthodontics. GH is an exaggerated increase in gingival tissue whose pathogenesis is unknown. However, it has been reported that the epithelial-mesenchymal transition as a process involved in other types of GH. Materials and methods: descriptive study that included the analysis of gingival tissues of healthy individuals (n = 6) and patients with GH by orthodontic treatment (n = 6). Before gingival surgery, the patients underwent a periodontal hygiene phase. The gingival tissue samples obtained were processed and embedded in paraffin. The cuts were made with a microtome and deposited on polysine adhesion slides. Histological hematoxylin-eosin staining was performed. The identification and location of S-100A4 and α-SMA markers was determined by immunohistochemistry with monoclonal antibodies. The reading of the findings was carried out by oral pathologists. Results: in healthy individuals, an S100A4 label was observed in Langerhans cells, while α-SMA was identified in the vascular endothelium of all samples analysed. However, in patients with GH due to orthodontics, they registered an intense staining of S100A4 in gingival fibroblasts, Langerhans cells, vascular endothelium, and areas adjacent to the rupture of blood vessel. α-SMA expression in GO was detected in the vascular endothelium and gingival fibroblasts. Conclusion: the differential immunostaining of EMT markers in gingival tissues of patients with orthodontic GH suggests an eventual role of EMT in the pathogenesis of this pathology..Au

S100a4 promotes differentiation of neural stem cells through up-regulation of brain-derived neurotrophic factor / 中国组织工程研究

BACKGROUND: How to promote neural stem cells differentiate into neurons is a difficulty. S100A4 has been found to play a role in the nervous system repair by various pathways. OBJECTIVE: To investigate whether S100A4 affects the differentiation of neural stem cells into neurons through up-regulating the expression of brain-derived neurotrophic facto. METHODS: The neural stem cells from brain hippocampus and subependymal region of embryonic mice were cultured in vitro and passaged. The S100A4 expression vector and/or brain-derived neurotrophic factor + siRNA were transfected into neural stem cells by electroporation, and the cells were induced to differentiate into neurons at 48 hours after transfection. Three days later, the expression levels of brain-derived neurotrophic factor and Tuj1 in cells were detected by western blot assay. Proportion of Tuj1 positive neurons was tested by immunofluorescence. RESULTS AND CONCLUSION: Compared with the unrelated sequence plasmid group, the proportion of Tuj1 positive neurons and the expression levels of Tuj1 and brain-derived neurotrophic factor in the S100A4 transfection group were significantly increased (P < 0.01). Compared with the S100A4+siRNA unrelated sequence plasmid group, the proportion of Tuj1 positive neurons and the expression levels of Tuj1 and brain-derived neurotrophic factor in the co-transfection group were significantly decreased (P < 0.01). These results indicate that S100A4 overexpression can promote the differentiation of neural stem cells into neurons, which may be mediated by brain-derived neurotrophic factor.

Benzyl isothiocyanate inhibits invasion and induces apoptosis via reducing S100A4 expression and increases PUMA expression in oral squamous cell carcinoma cells

Benzyl isothiocyanate (BITC) has been shown to inhibit invasion and induce apoptosis of various types of cancer. However, its role on human oral squamous cell carcinoma (OSCC) cells is still not well elucidated. In the present study, we investigated the effect of BITC on apoptosis and invasion of SCC9 cells, and its underlying mechanisms in vitro and in vivo. SCC9 cells were exposed to BITC (5 and 25 μM) for 24 and 48 h. Cell growth, apoptosis, invasion, and migration were detected in vitro by MTT, FITC-conjugated annexin V/propidium iodide staining followed by flow cytometry, Matrigel-coated semi-permeable modified Boyden, and wound-healing assay. S100A4, PUMA, and MMP-9 expressions were detected to investigate its mechanisms. Xenotransplantation experiments were used to investigate the role of BITC on tumor growth and lung metastasis. BITC inhibited cell viability and induced cell apoptosis in a dose- and time-dependent manner through upregulation of PUMA signals. BITC inhibited cell invasion and migration by downregulation of S100A4 dependent MMP-9 signals. The ip administration of BITC reduced tumor growth but not lung metastasis of SCC9 cells subcutaneously implanted in nude mice. BITC treatment activated pro-apoptotic PUMA and inhibited S100A4-dependent MMP-9 signals, resulting in the inhibition of cell growth and invasion in cultured and xenografted SCC9 cells. Thereby, BITC is a potential therapeutic approach for OSCC.

Expression of metastasis associated genes nm23, p53 and S100A4 in conjunctival melanoma and their relationship with invasion and metastasis / 中华实验眼科杂志

Objective To study the significance of metastasis associated genes nm23,p53 and S100A4 in the development of conjunctival melanoma metastasis.Methods Conjunctival melanoma tissue specimens were collected from 42 cases of conjunctival melanoma patients in Henan Eye Hospital from July 2015 to November 2016,meanwhile 30 cases of conjunctival nevus tissue samples served as control group under the informed consent.The expressions of nm23,p53 and S100A4 were detected in conjunctival melanoma group and control group by Western blot and immunohistochemical method.The relationship between the clinical and pathological features of nm23,p53 and S100A4 with conjunctival melanoma patients with lymph nodes metastasis were analyzed.Results Western blot assay showed that the expression level of nm23 in conjunctival melanoma group was lower than that in the control group,with a significant difference between them (P＜0.05);the expression level of p53 and S100A4 in conjunctival melanoma group was significantly higher than those in the control group (both at P＜0.05).Immunohistochemistry showed that nm23 and S100A4 appeared claybank in cytoplasm,while p53 appeared red in cell nucleus.The positive rate of nm23 protein expression in conjunctival melanoma group was significantly lower than that in the control group (14.3％ vs.53.3％,P＜0.05).The positive rate of p53 expression in conjunctival melanoma group was significantly higher than that in the control group (54.8％ vs.6.7％,P＜0.05).The positive rate of S100A4 expression in the conjunctival melanoma group was significantly higher than that in the control group (59.5％ vs.6.7％,P＜0.05).There were no difference in nm23,p53 and S100A4 protein expression positive rate between various gender groups,various age groups and various ulcer groups (all at P＞0.05).There were siginificant differences in nm23,p53 and S100A4 protein expression positive rate between various sclera invasion groups and various distant metastasis groups (all at P＜0.05).Conclusions Expression of nm23,p53 and S100A4 may play an important role in the invasion and metastasis of conjunctival melanoma,which may be the main reference index for the diagnosis and treatment.

Effects of Elemene Combined with Chemotherapy on S100A4 and MMP-9 Protein Expression in Lewis Lung Carcinoma Mice / 广州中医药大学学报

Objective To investigate the effects of elemene combined with chemotherapy on S100A4 and MMP-9 protein expression in Lewis lung carcinoma mice and to explore the possible mechanism. Methods Twenty-four mice inoculated with Lewis cells were randomly divided into 4 groups,including lung carcinoma model group,chemotherapy group,elemene group,and combination group,6 mice in each group. On day 11 after inoculation, the mice in chemotherapy group were given intraperitoneal injection of etoposide (3 mg·kg-1·d-1)and cisplatin (3 mg·kg-1·d-1),the mice in elemene group were given intraperitoneal injection of elemene (100 mg·kg-1·d-1), the mice in the combination group were given intraperitoneal injection of elemene (100 mg·kg-1·d-1),etoposide (3 mg·kg-1·d-1),and cisplatin (3 mg·kg-1·d-1). After 7-day continuous treatment,the tumor body mass, spleen index, thoracic gland index, growth-inhibition rate, and metastasis-inhibition rate in various groups were measured,the contents of protein S100A4 and matrix metalloproteinase 9 (MMP-9)in tumor tissues were determined by enzyme-linked immunosorbent assay (ELISA),and the protein expression levels of S100A4 and MMP-9 were detected by immunohistochemical staining. Results In the combination group, the growth-inhibition rate and the inhibitory rate of metastases were increased obviously, the spleen index and thoracic gland index were also increased, the contents of S100A4 and MMP-9 were decreased, and the protein expression rates of S100A4 and MMP-9 were reduced(P < 0.05 or P < 0.01 compared with the chemotherapy group). Conclusion Elemene combined with chemotherapy could inhibit the Lewis lung carcinoma growth and metastasis in mice, and the possible mechanism might be associated with the decrease of S100A4 and MMP-9 protein expression in the tumor tissues.

Expression of vascular endothelial growth factor,S100A4 and survivin in thyroid carcinoma and their clinical significance / 中国基层医药

Xu XP,Xiao N);Department ofGeneral Surgery,Pingxiang People's Hospital,Pingxiang,Jiangxi 337000,China(Rong J) Objective To study the expression of vascular endothelial growth factor(VEGF),S100A4 and survivin in thyroid carcinoma and their clinical significance.Methods The expression of VEGF,S100A4 and survivin in normal thyroid tissues from 28 people and thyroid cancer tissues obtained from 73 patients were detect by immuno-histochemistry method.Results The positive rates of VEGF,S100A4 and survivin in thyroid cancer were 84.93%, 86.30% and 79.45%,which in the normal thyroid tissues were 3.57%,7.14% and 14.29%,respectively.There were significant differences between every two groups,thyroid cancer and the normal thyroid tissues,statistically(χ2 =57.08,55.28,36.26,all P =0.000).VEGF,S100A4 and survivin in the positive expression of capsular infiltration carcinoma group were 97.37%,97.74%,97.74%,which in lymph node metastasis positive group cancer rates were 97.30%,94.60%,97.30% were significantly increased compared with no capsular infiltration group (71.43%, 77.14%,62.86%,respectively)and lymph node metastasis group(72.22%,77.78%,61.11%),the differences were statistically significant(χ2 =8.96,4.37,14.63,9.58,4.77,11.34,all P <0.05);the positive expression rate of three with thyroid cancer histological grade increased.Conclusion VEGF,S100A4 and survivin are closely related to the aggressive and metastatic character of thyroid cancer.A combination of VEGF,S100A4 and survivin may be of grate significance for predicting high aggressive and metastatic character in thyroid carcinoma.

Effect of small interference RNA silenced S100A4 protein in human gastric carcinoma cells on proliferation, apoptosis and chemotherapy sensitivity in vitro / 国际外科学杂志

Objective To study the application of small interfering RNA silencing S100A4 protein in human gastric cancer cell BGC-823 proliferation,apoptosis and the effect of chemotherapy sensitivity.Methods Human gastric carcinoma cell line BGC-823 transfection siRNA,RT-PCR detected the changes of mRNA after transfection.Groups divided into interference group,negative control group and normal control group.MTT test determined different concentrations of oxaliplatin in gastric cancer cells and calculated IC50,then draw cell growth curve,TUNEL method to detect apoptosis,RT-PCR tested each cell mRNA changed,Western blot detected the change of the S100A4 protein.All data analysis by SPSS17.0,t test applied,RT-PCR and Western blot results analysis by SPSS17.0,comparing multiple samples by using single factor analysis of variance and LSD test.P ＜ 0.05 was statistically significant.Results RT-PCR results showed that BGC-823 cell transfection,S100A4mRNA expression quantity respectively after 48 hours:(0.674+0.011),(0.652+0.021),(0.345 + 0.040),the interference group and normal control group were statistically significant (P =0.012,P ＜ 0.05) and the negative control group with interference group differences were statistically significant (P =0.000,P ＜ 0.05),and normal control group was no statistically significant difference with the negative control group (P =0.380,P ＞ 0.380);Western blot results showed BGC-823 cell transfection S100A4 expression significantly lowered respectively after 48 hours,there were (0.654 + 0.025),(0.642 + 0.014),(0.317 ± 0.061),the interference group and normal control group was statistically significant (P =0.01,P ＜ 0.05),between negative control group and interference group were statistically significant (P =0.000,P ＜ 0.05),normal control group and the negative control group had no significant difference (P =0.341,P ＞ 0.341).After S100A4-siRNA transfection,gastric carcinoma BGC-823 cell proliferation decreased,TUNEL method showed obviously increase apoptosis,MTY showed that IC5o of oxaliplatin was 56.31 μmol/L,after transfection,IC50 was 0.654 μmol/L.Conclusions This study showed that the siRNA silence S100A4 protein inhibit gastric cancer cell proliferation,induced apoptosis and improved chemotherapy sensitivity of oxaliplatin.S100A4 might be prompt targets for the treatment of gastric carcinoma.

Dose-response of S100A4 gene expression in human lymphocytes at different time points after irradiation / 中华放射医学与防护杂志

Objective To investigate the dose response of S100A4 gene expression in the irradiated lymphoblastoid cells AHH-1 at different time points post irradiation.Methods AHH-1 cells was exposed to different doses(0,1,3,5,8,10,15 and 18 Gy)of 60Co γ-rays,and its mRNA levels of S100A4 was detected by reverse transcription PCR and real-time PCR at 4,8,12,24,48 and 72 h after irradiation.Results Within the range of applied doses,the level of S100A4 gene expression was upregulated with a good dose-response (R2 =0.79-0.93,P ＜ 0.05) and had obvious difference at different time points (F =8.91,P ＜ 0.01).Conclusion S100A4 gene expression at transcriptional level could be detected easily and had optimum dose-responses at certain time points after irradiation,and hence is applicable as a dosimeter.

The effects of knockdown of S100A4 on invasion and migration of SNB19 glioma cells / 中国神经精神疾病杂志

Objective To investigate the effects of siRNA-mediated knockdown of S100A4 expression on the inva?sion and migration of SNB19 glioma cells. Methods The S100A4 expression was knockdowned using S100A4 siRNA in SNB19 glioma cells. Glioma cells were assigned into control group,siRNA-negative control treated group (siRNA-NC) and siRNA-S100A4 group. RT-PCR and western blot were used to detect the mRNA and protein expression of S100A4, respectively. The wound-healing assay and transwell invasion assay were used to determine the ability of migration and invasion of SNB19 glioma cells, respectively. The expression of matrix metalloproteinase 9 (MMP-9), matrix metallopro?teinase 2 (MMP-2) and E-cadherin proteins were evaluated by using western blot. Moreover, the morphology of lamellipo?dia of glioma cells were examined by using inverted phase-contrast microscopy. Results The mRNA and protein expres?sion levels of S100A4 was obviously down-regulated after transfection of S100A4 siRNA. Compared with control group, the mRNA expression levels of S100A4 in siRNA-NC group and siRNA-S100A4 group were 0.97±0.07 and 0.21±0.04,respectively(P<0.01). The protein expression levels of S100A4 in control, siRNA-NC and siRNA-S100A4 groups were 78.12%±2.63%, 77.16%±3.00%and 37.95%±2.71%, respectively(P<0.01). The migration and invasiveness capability were decreased up to 46% and 55% in the siRNA-S100A4 group compared with the control group(P<0.01). The pro?tein expression levels of MMP-9 and MMP-2 were inhibited up to 62% and 68%(P<0.01)whereas the expression of E-cadherin was increased up to 154%(P<0.01)in the siRNA-S100A4 group. The lamellipodia became smaller or unex?tended in siRNA-S100A4-treated SNB19 glioma cells. Conclusion S100A4 plays an important role in the invasion and migration of glioma cells, suggesting that S100A4 might be a potential candidate for anti-glioma strategy to prevent the invasion and migration of glioma cells.

The effect of S100A4 siRNA on the inflammation and expression of TNF-α,IL-1β in adjuvant arthritis rats / 重庆医学

Objective To observe the effects of S100A4 siRNA on the expression of serum TNF‐α,IL‐1βand VEGF in adjuvant arthritis rats .Methods Adjuvant arthritis rat models were established and were randomly divided into model group and interfere group .On Day 11 ,rats in interfere group were injected with S100A4 siRNA fragment in articular cavity .Arthritis index (AI) chan‐ges and pathological changes of ankle joint were observed .The levels of serum TNF‐α and IL‐1β ,VEGF were detected by ELISA . Results Compared with that of model group ,the levels of serum TNF‐ α ,IL‐1β and VEGF were reduced significantly in interfere group (P< 0 .05) ;variances of AI and pathological scores in interfere group were diminished significantly (P< 0 .05) .Conclusion Inhibition of the expression of S100A4 gene can significantly reduce the expression of inflammatory factor TNF‐α ,IL‐1 β and angio‐genesis factor VEGF ,and improve the pathological injury of synovial membrane .

Effect of S100A4 silencing on tumor related gene mRNA expression / 中华胰腺病杂志

Objective To investigate the effect of S100A4 silencing on tumor related gene COX-2,bcl-2,Surviving,MMP-9 mRNA expressions of pancreatic cancer BxPC-3,AsPC-1 cells,and explore their relationship.Methods Small interfering RNA interfering S100A4 gene (siRNA-S100A4) was applied to transfect human pancreatic cancer BxPC-3,AsPC-1 cells,and nonhomologous siRNA-C was used as negative control,and cells without transfection were used as control group.The expressions of S100A4,COX-2,Survivin,MMP-9,bcl-2 mRNA after interference were detected by using RT-PCR.Results S100A mRNA expressions of BxPC-3's control group,siRNA-C group,siRNA-S100A4 group were 0.661 ± 0.023,0.659 ± 0.043,0.379 ± 0.039,and expressions of COX-2 mRNA were 0.760 ± 0.026,0.830 ± 0.017,0.443 ±0.006,and expressions of Survivin mRNA were 0.948 ± 0.049,0.909± 0.081,0.068 ± 0.006,and expressions of bcl-2 mRNA were 0.462 ±0.018,0.421 ±0.049,0.184 ±0.025,and expressions of MMP-9 mRNA were 0.813 ± 0.008,0.908 ± 0.063,0.246 ± 0.027.S100A mRNA expressions of AsPC-I's control group,siRNA-C group,siRNA-S100A4 group were 0.641 ± 0.042,0.626-± 0.053,0.320 ± 0.081,and expressions of COX-2 mRNA were 0.727 ± 0.021,0.743 ± 0.025,0.560 ± 0.035,and expressions of Survivin mRNA were 0.994 ± 0.032,0.984 ± 0.049,0.063 ± 0.005,and expressions of bcl-2 mRNA were 0.458 ±0.004,0.537 ± 0.046,0.181 ± 0.007; and expressions of MMP-9 mRNA were 0.698 ± 0.011,0.718 ± 0.073,0.199± 0.013.The expressions of S100A,COX-2,Survivin,bcl-2,MMP-9 mRNA in groups with siRNA-S100A4 transfection were significantly lower than those of siRNA-C group and control group (P ＜0.01),but the difference between siRNA-C group and control group was not statistically significant.Conclusions S100A4 plays a role in the pathogenesis of pancreatic cancer through up-regulation of COX-2,Survivin,bcl-2,MMP-9 expressions.

Correlation of Immunohistochemical Markers and BRAF Mutation Status with Histological Variants of Papillary Thyroid Carcinoma in the Korean Population

Several pathologic characteristics are associated with an adverse clinical outcome in papillary thyroid carcinoma (PTC), including the histological variant. This study aimed to investigate immunohistochemical expression and BRAF mutation status based on the histological variant and evaluated potential markers of aggressive behavior of PTC in Korean patients. In all, 407 PTC cases were classified to each histological variant, and the 94 representative cases were subjected to immunohistochemistry and BRAF mutation analysis. The classic type, follicular variant (FV) and tall cell variant (TCV) represented 76.9%, 14.2% and 6%, respectively. TCV showed a larger tumor size (P = 0.009), frequent extrathyroidal extension (P = 0.022) and cervical lymph node (LN) metastasis (P = 0.018). TCV and FV showed the reduced expression of galectin-3 (P = 0.003) and HBME1 (P = 0.114). Regardless of histology, PTEN loss and diffuse S100A4 expression were associated with LN metastasis (P = 0.007, P = 0.013). All TCVs harbored BRAF V600E mutation, and FV harbored less BRAF V600E mutation (P = 0.043). Immunohistochemical evaluation showed characteristic patterns in histological variants. PTEN and S100A4 expression are suggested as indicators of regional lymph node metastasis.

Podoplanin, alpha-Smooth Muscle Actin or S100A4 Expressing Cancer-Associated Fibroblasts Are Associated with Different Prognosis in Colorectal Cancers

The interactions between the tumor microenvironment and tumor cells determine the behavior of the primary tumors. Whether cancer-associated fibroblasts (CAF) have a tumor progressive or a protective role likely depends on the type of tumor cells and the CAF subpopulation. In the present study, we analyzed the prognostic significance of CAF subpopulations in colorectal cancer (CRC). CAF phenotypes were analyzed in 302 CRC patients by using antibodies against podoplanin (PDPN), alpha-smooth muscle actin (alpha-SMA), and S100A4. The relationship between the CAF phenotypes and 11 clinicopathological parameters were evaluated and their prognostic significance was analyzed from the disease-free and overall survival times. We observed that at the tumor invasive front, PDPN CAFs were present in 40% of the cases, and S100A4 or alpha-SMA CAFs were detected in all the cases. PDPN/S100A4 and alpha-SMA/S100A4 dual-stained CAFs were observed in 10% and 40% of the cases, respectively. The PDPN+ CAFs were associated with 6 favorable clinicopathological parameters and prolonged disease-free survival time. The PDPN-/alpha-SMA(high) CAFs were associated with 6 aggressive clinicopathological parameters and tended to exhibit shorter disease-free survival time. On the other hand, the PDPN-/S100A4(high) CAFs were associated with 2 tumor progression parameters, but not with disease prognosis. The PDPN+ CAF phenotype is distinct from the alpha-SMA or S100A4 CAFs in that it is associated with less aggressive tumors and a favorable prognosis, whereas the PDPN-/alpha-SMA(high) or PDPN-/S100A4(high) CAFs are associated with tumor progression in CRC. These findings suggest that CAFs can be a useful prognostic biomarker or potential targets of anti-cancer therapy in CRC.

Correlation of Cav-1 mRNA, S100A4 mRNA and CD31 expression with prostate cancer tumor metastasis and patient survival rate / 中华泌尿外科杂志

Objective　To explore the expression of Cav-1 mRNA,S100A4 mRNA and CD31 in prostate cancer (PCa) and their correlation with tumor metastasis and patient survival rate.　Methods PCa specimens (n =42) and adjacent tissue specimens ( n =12 ) from radical prostatectomy were obtained from January 2004 to May 2006.The mean age of patient was 71.6 ± 7.6 years ( range 58 - 86 years).According to Gleason scores,prostatectomy specimens were stratified into≤6 (n =17),7 (n =12) and ≥8( n =13 ) groups.Patients were classified as clinical stage T1 ( n =16),stage T2 ( n =9 ),stage T3 ( n =11 )and stage T4 (n =6).Patients were divided into PCa with bone metastasis (n =8 ) and PCa without bone metastasis ( n =34).Preoperative PSA levels of the patients were stratified into three groups: ＜ 4 ( n =4)μg/L,4-10 (n=10)μg/Land ＞10 μg/L (n=28).12 adjacent tissues 1 -2 cm away from tumor or another lobe of prostate were microscopically verified without cancer cells and were tested for comparison.The expression of Cav-1 mRNA and S100A4 mRNA were detected by Situ hybridization in 42 PCa specimens and 12 adjacent tissues and using CD31 for marking vascular endothelial cells,the tumor microvascular density (MVD) was counted.The correlation of Cav-1 mRNA,S100A4 mRNA and CD31 expression was analyzed in combination.with clinical and pathological fcatures including Gleason score,TNM staging,PSA values and bone metastasis.　Results　The positive expression rate of Cav-1 mRNA in PCa was 35.7％　( 15/42),while it was 0％ (0/12) in controls,P ＜0.05.The positive expression rate of S100A4 mRNA in PCa was 47.6％ (20/42),while it was 8.3％ (1/12) in controls,P ＜0.05.The positive expression rate of Cav-1 mRNA in PCa was positively correlated with Gleason score,TNM stage and bone metastasis.The positive expression rate of S100A4 mRNA in PCa was positively correlated with TNM stage and bone metastasis.The average MVD in patients of negative expression of Cav-1 mRNA was (62.8 ± 10.4)/mm2,and the average -MVD in patients of positive expression of Cav-1 mRNA was (83.5 ±6.7 )/mm2,P ＜ 0.05.While the average MVD in patients of negative expression of S100A4 mRNA was (63.3 ± 12.0)/mm2,and the average MVD in patients of positive expression of S100A4 mRNA was (77.9 ± 11.0)/mm2,P ＜ 0.05.The 5-year survival rate in patients with positive Cav-1 mRNA expression was significantly lower than that of with negative expression (46.7％ versus 85.2％,P ＜ 0.05 ),while the 5-year survival rate in the patients with positive expression of S100A4 mRNA was significantly lower than that of with negative expression (50.0％ versus 90.9％,P ＜ 0.05 ).　Conclusions　The positive Cav-1 mRNA and S100A4 mRNA expression,increased MVD are positively correlated with PCa progression and bone metastasis. Furthermore,Cav-1 and S100A4 in PCa may promote angiogenesis and cause tumor cells to bone metastases,which can reduce survival rate of patients.

Expressions of S100A4 and uPA and the correlation with pancreatic cancer prognosis / 国际肿瘤学杂志

Objective To study the expressions of S100A4 and urokinase plasminogen activator(uPA) in pancreatic cancer cells and their correlation with patients prognosis.Methods The expressions of S100A4 and uPA were examined in 63 surgical specimens of primary pancreatic carcinoma by suing immunohistochemistry PV methods,and correlation of their expressions and prognosis of pancreatic cancer was analyzed.Results ( 1 ) Positive immunostaining for S100A4 and uPA was observed in 74.6％ (47 cases) and 65.1％ (44 cases) of 63 pancreatic cancer samples respectively.(2) The positive expressions of S100A4 and uPA were significantly correlated in pancreatic cancer( P =0.000,r =0.567 ).( 3 ) The expression of S100A4 significantly correlated with TNM stages( P =0.002 ),lymph node metastasis ( P =0.002 ) and distant metastasis ( P =0.007 ).The expression of uPA had a significant correlation with TNM stages ( P =0.002),lymph node metastasis ( P =0.001 )and differentiation(P =0.003).(4) Kaplan-Meier survival analysis showed that the median survival (21 months) of patients with S100A4 ( - ) was significantly longer than the median survival ( 9 months) of the patients with S100A4( + )(P=0.000) ;the median survival(9 months) of patients with uPA( + ) was significantly shorter than the median survival ( 18 months) of the patients with uPA ( - ) ( P =0.000) ; the median survival(23 months)of 13 patients with S100A4( - )/uPA( - )was significantly longer than the median survival of other cases ( Log-rank test,x2 =54.444,P =0.000).( 5 ) Cox regression model ( x2 =53.974,P =0.000 )analysis suggested:the differentiation(P =0.004),lymph node metastasis(P =0.017) 、S100A4( + ) expression ( P =0.000) and uPA ( + ) expression ( P =0.001 ) were independent prognostic factors for pancreatic cancer.Conclusion S100A4 and uPA are highly expressed in pancreatic cancer cells,and S100A4 expression has positive correlation with uPA expression,which indicates that the overexpression of S100A4 and uPA maybe poor prognosis factors for pancreatic cancer patients.S100A4 maybe promote extracellular matrix and basement membrane degradation by up-regulation of uPA protein expression,and ultimately promote tumor invasion and metastasis,which is not favorable to prognosis.They may be potential indicators of metastasis and predictors for prognosis of pancreatic cancer.

Podocyte Expression of Osteopontin and FSP-1/S100A4 in Human Crescentic Glomerulonephritis

BACKGROUND: Osteopontin (OPN) is a cytokine associated with a cell-matrix via integrins. Fibroblast specific protein-1 (FSP-1), known as S100A4, has been implicated in cell migration by non-muscle myosin. We investigated whether the role of OPN and FSP-1/S100A4 expression in their contribution to the podocyte phenotype change to form podocyte bridge and cellular crescent. METHODS: Glomerular expression of OPN and FSP-1/S100A4 in renal biopsies of 16 patients with crescentic glomerulonephritis (CrGN) and 13 normal renal biopsies were studied by immunohistochemistry. RESULTS: The expression of OPN and FSP-1/S100A4 was increased in the podocytes of glomeruli, with and without crescents, in patients with CrGN. Neither OPN nor FSP-1/S100A4 was expressed in glomeruli from the normal controls (p<0.01). A significant positive correlation was found between the expression of OPN in glomerular tufts and cellular crescents, and the expression of OPN and FSP-1/S100A4 in glomerular tufts (p<0.05). CONCLUSIONS: The results suggest that OPN plays a role in early podocyte attachment to Bowman's capsule, and FSP-1/S100A4 potentiate podocyte contribution to cellular crescent formation by inducing cellular migration and growth.

Study on S100A4 gene silence by siRNA and its inhibition on invasion of human thyroid carcinoma cell / 中华内分泌外科杂志

Objective To explore the effects and mechanism of S100A4 gene silence on invasion of human thyroid cancer cell. Methods After thyroid cancer cell ARO was transfected by S100A4 small interfering RNA (siRNA), mRNA and protein level of S100A4 and matrix metalloproteinase 2 (MMP-2) were determined by real time RT-PCR and Western blot respectively. The anchorage-independent growth was examined by colony formation assay in soft agar, and invasion ability was evaluated by boyden chamber model. Results The level of mRNA and protein of S100A4 was significantly inhibited in ARO cancer cells transfected by S100A4 siRNA.Transfection with S100A4 siRNA could inhibit anchorage-independent growth and invasion ability of thyroid cancer cell ARO in a dose-dependent manner. mRNA and protein expression of MMP-2 were down-regulated by S100A4 siRNA. Conclusion S100A4 siRNA can inhibit the invasion of thyroid cancer cell through down-regulation of MMP-2.

Significance of S100A4 expression in benign and malignant thyroid tissue / 中华内分泌外科杂志

Objective To investigate the expression of S100A4 protein in benign and malignant thyroid tissues and its clinical significance. Methods Expression of S100A4 protein in thyroid cancers (50 cases), adenoma (45 cases) and adjacent noncancerous tissue (20 cases) was respectively studied by microwave-LSAB immunohistochemical staining. Results The positive rate of S100A4 protein in thyroid cancer and adenoma was 78.0% and 15.6% respectively. No S100A4 protein expression was found in any adjacent noncancerous tissue (P ＜0. 01). No correlation was observed between expression of S100A4 protein and histological grading of thyroid cancer. Expression of S100A4 protein in lymph node metastasis and staging Ⅲ-Ⅳ was both higher than that in negative cases and staging Ⅰ -Ⅱ (P ＜0. 05). The rate of recurrence and death in S100A4 protein positive cases were notably higher than that in negative cases (P ＜ 0. 05). Conclusions S100A4 protein expression can help to distinguish benign and malignant thyroid tissues. It can be used as a molecular marker to predict thyroid metastasis.

A study on the expression of BCSG1, S100A4, VEGF in breast neoplasm / 中华普通外科杂志

Objective To study the expression of BCSG1 ,S100A4,VEGF in human breast cancer.Methods Immunohistochemistry technique SP method was used to examine breast fibroadenoma in 40 cases,breast invasive ductal carcinoma in 62 cases ( by armpit lymph node metastasis as 2 groups) and corresponding paratumor tissues in 48 cases. Results Among the 4 groups, there was significant difference ( P＜0.05 ) in the positive immunostaining rate of BCSG1 ,S100A4, VEGF. Conclusions In the breast invasive ductal carcinoma, the expression of BCSG1, S100A4 and VEGF increased. This suggested the invasive and metastasis ability of the neoplasm enhanced. The expression were in positive correlation with tumor pathology. Combined detection of BCSG1, S100A4, VEGF expression contributes to the early diagnosis and prognostic assessment of the carcinoma of the breast.