EF手型钙结合蛋白S100A6及其与肿瘤关系的研究进展

S100A6蛋白是一类具有EF手型模序结构的钙结合蛋白,通过对钙离子的调节及与靶蛋白的相互作用,启动细胞信号转导,参与细胞生长、分化、细胞周期及胞外基质分泌等多种生物学过程,与肿瘤发生、发展及转移预后相关.本文就近年来S100A6钙结合蛋白的研究现状与进展进行综述.     

c-FLIP confers resistance to FAS-mediated apoptosis in anaplastic large-cell lymphoma

We hypothesized that inhibition of the FAS-mediated apoptosis pathway by FLICE-like inhibitory protein (c-FLIP) may contribute to oncogenesis in ALK+ anaplastic large-cell lymphoma (ALCL). Treatment with increasing concentrations of CH-11 (CD95/FAS agonistic antibody) had no effect on cell viability of 2 ALK+ ALCL cell lines, Karpas 299 and SU-DHL1, each expressing high levels of c-FLIP. However, inhibition of endogenous c-FLIP expression by specific c-FLIP siRNA in Karpas 299 and SU-DHL1 cells treated with CH-11 resulted in FAS-mediated cell death associated with increased annexin V binding, apoptotic morphology, and cleavage of caspase-8. In 26 ALK+ ALCL tumors, assessed for expression of DISC-associated proteins, CD95/FAS and c-FLIP were commonly expressed, in 23 (92%) of 25 and 21 (91%) of 23 tumors, respectively. By contrast, CD95L/FASL was expressed in only 3 (12%) of 26 ALCL tumors, although it was strongly expressed by surrounding small reactive lymphocytes. Our findings suggest that overexpression of c-FLIP protects ALK+ ALCL cells from death-receptor-induced apoptosis and may contribute to ALCL pathogenesis.     

Colorectal cancer progression correlates with upregulation of S100A11 expression in tumor tissues

Background and aims Early detection and treatment of human colorectal cancers remain a challenge. Identification of new potential markers may help in the diagnosis of colorectal cancer. Materials and methods By comparative two-dimensional gel electrophoresis using extracts from colorectal tumor and adjacent normal tissues, we identified a calcium-binding protein, S100A11, which was highly expressed in colorectal cancer compared with adjacent normal tissues. We expanded our study in 89 clinical colorectal tumor samples to validate this finding and correlates S100A11 expression in human colorectal cancer tissues with various stages of the tumor by Western blotting and immunohistochemical staining. Results We identified a calcium-binding protein, S100A11, which was highly expressed in colorectal cancer compared with adjacent normal tissues. S100A protein was expressed predominantly in the cytoplasm of normal tissue; however, it was expressed in both the nuclei and cytoplasm of colorectal cancer. S100A11 level in colorectal cancer tissue was increased following stage progression of the disease. Conclusion These findings suggest S100A11 could be helpful in the pathological study of colorectal cancer, especially for the classification of different stages in colorectal cancer.     

Loss of SHP1 enhances JAK3/STAT3 signaling and decreases proteosome degradation of JAK3 and NPM-ALK in ALK+ anaplastic large-cell lymphoma

Previous studies showed that most cases of ALK(+) anaplastic large-cell lymphoma (ALK(+)ALCL) do not express SHP1, a tyrosine phosphatase and an important negative regulator for cellular signaling pathways such as that of JAK/STAT. To fully assess the biologic significance of loss of SHP1 in ALK(+)ALCL, we transfected SHP1 plasmids into 2 SHP1(-), ALK(+)ALCL cell lines, Karpas 299 and SU-DHL-1. After 24 hours of transfection, pJAK3 and pSTAT3 were decreased, and these changes correlated with down-regulation of STAT3 downstream targets including cyclin D3, mcl-1, and bcl-2. Expression of SHP1 in these 2 cell lines also resulted in marked decreases in the protein levels of JAK3 and NPM-ALK, and these effects were reversible by proteosome inhibitor MG132. Conversely, when SHP1 expression in SUP-M2 (a SHP1(+) ALK(+)ALCL cell line) was inhibited using siRNA, pSTAT3, pJAK3, JAK3, and NPM-ALK were all up-regulated. Coimmunoprecipitation studies showed that SHP1 was physically associated with JAK3 and NPM-ALK. SHP1 expression in Karpas 299 and SU-DHL-1 led to significant G(1) cell cycle arrest but not apoptosis. To conclude, loss of SHP1 contributes to the pathogenesis of ALK(+)ALCL by 2 mechanisms: (1) it leaves the tyrosine phosphorylation and activation of JAK3/STAT3 unchecked and (2) it decreases proteosome degradation of JAK3 and NPM-ALK.     

Gene-expression profiling of systemic anaplastic large-cell lymphoma reveals differences based on ALK status and two distinct morphologic ALK+ subtypes

With the use of microarray gene-expression profiling, we analyzed a homogeneous series of 32 patients with systemic anaplastic large-cell lymphoma (ALCL) and 5 ALCL cell lines. Unsupervised analysis classified ALCL in 2 clusters, corresponding essentially to morphologic subgroups (ie, common type vs small cell and "mixed" variants) and clinical variables. Patients with a morphologic variant of ALCL had advanced-stage disease. This group included a significant number of patients who experienced early relapse. Supervised analysis showed that ALK+ALCL and ALK- ALCL have different gene-expression profiles, further confirming that they are different entities. Among the most significantly differentially expressed genes between ALK+ and ALK- samples, we found BCL6, PTPN12, CEBPB, and SERPINA1 genes to be overexpressed in ALK+ ALCL. This result was confirmed at the protein level for BCL-6, C/EBPbeta and serpinA1 through tissue microarrays. The molecular signature of ALK- ALCL included overexpression of CCR7, CNTFR, IL22, and IL21 genes but did not provide any obvious clues to the molecular mechanism underlying this tumor subtype. Once confirmed on a larger number of patients, the results of the present study could be used for clinical and therapeutic management of patients at the time of diagnosis.     

Upregulated expression of S100A6 in human gastric cancer

OBJECTIVE: The expression of S100A6 (calcyclin), a member of the S100 calcium binding protein family, is elevated in a number of malignant tumors, but there have been few reports about its expression in gastric cancer. The aim of this study was to investigate its expression regulations in human gastric cancer and noncancerous mucosa, and the response to chemotherapeutic drugs in the gastric cancer cell line. MATERIALS AND METHODS: In one matched gastric cancer sample pair, the serial analysis of gene expression (SAGE) experiment was conducted to compare the gene expression profiles between cancerous and adjacent tissues. To detect the expression regulations among more cancerous tissues, microarrays were carried out and real-time RT-PCR was conducted to validate the results. At the protein level, Western blot and tissue microarray (TMA) examination were further used to verify S100A6 expression. The regulation detection of S100A6 with flurouracil and doxorubicin at the mRNA and protein level was performed in the SGC7901 cell line. RESULTS: With the SAGE strategy, five times more S100A6 tags were identified in cancer tissues than in normal tissues. With the cDNA microarray, S100A6 was found to be significantly upregulated in 21 of 42 (50%) nonselective gastric cancers. In 10 other paired samples, the upregulation of S100A6 was consolidated with RT-PCR and Western blot analysis as well. A total of 14 endoscopy-sectioned gastric noncancerous lesions and corresponding normal gastric mucosa were also applied to profile the gene expression; both cDNA microarray and RT-PCR demonstrated no significant alterations of S100A6 at the mRNA level. TMA examination showed that 34 of 52 (65.4%) cancer samples were positively stained, while only 17 of 80 (21.3%) noncancerous lesions were positively detected and all nine normal mucosae were detected to be negative. An in vitro experiment showed that in the gastric cell line SGC-7901, S100A6 mRNA was detected to be upregulated from 24 to 72 h after treatment with 5 mg/L 5-flurouracil or 0.3 mg/L doxorubicin, and there were two wave upregulations of the S100A6 protein. CONCLUSION: The observed regulated expression of S100A6 suggests that it is associated with gastric cancer tumorigenesis and quantitation of S100A6 is a promising tool for diagnosis of gastric cancer.     

非小细胞肺癌患者S100A4蛋白表达及其临床意义

目的探讨非小细胞肺癌S100A4蛋白的表达及临床意义。方法选择130例非小细胞肺癌患者术后癌组织标本,用免疫组织化学S-P方法,检测NSCLC中的S100A4蛋白含量并进行统计学分析。结果在NSCLC中S100A4蛋白阳性率为69．2％;鳞状细胞癌、腺癌、腺鳞癌、大细胞肺癌阳性率分别为62．7％、68．1％、70．O％、100％;S100A4蛋白表达阳性表达与年龄、淋巴结转移、远处转移及TNM临床分期明显相关;S100A4蛋白阳性组和阴性组3年生存率比较有显著差异（P〈0．05）。结论S100A4蛋白的表达和与年龄、淋巴转移、临床分期及预后有密切相关;可作为肺癌早期的标记物。     

Hammerhead ribozyme-mediated cleavage of the fusion transcript NPM-ALK associated with anaplastic large-cell lymphoma

OBJECTIVE: Approximately 60% of all anaplastic large-cell lymphomas (ALCL) contain a specific t(2;5)(p23;q35) chromosomal translocation leading to overexpression of NPM-ALK. As the chimeric tyrosine kinase is involved in tumorigenesis and pathogenesis of ALCL, we were interested to inhibit NPM-ALK expression using an exogenous and an endogenous ribozyme approach. METHODS: We designed five anti-ALK hammerhead ribozymes that were targeted to cleave the ALK proportion of NPM-ALK. The ribozyme with the highest cleavage activity was used as a modified RNA/DNA chimera (RZ1*) for transient transfection and as a self-splicing ribozyme vector (pRZ1) for endogenous expression. Ribozyme performance was tested in 293 cells (cotransfected with NPM-ALK) and in the ALCL cell line Karpas 299 by transient and stable transfection and Western blotting. The half-life time of NPM-ALK was determined by pulse-chase experiments. RESULTS: In vitro cleavage assays demonstrated different catalytic efficiencies depending on the targeted site of the substrate. Constant transfection of Karpas 299 cells with RZ1* for 96 hours did not lead to a significant reduction of NPM-ALK protein, presumably due to the long half-life of NPM-ALK (48 hours). In contrast, NPM-ALK protein expression was almost completely suppressed in transiently transfected 293 cells. Stable transfection of Karpas 299 cells with pRZ1 also resulted in significant reduction of NPM-ALK expression. CONCLUSION: These results suggest that ribozymes targeted against NPM-ALK are able to inhibit expression of this oncogenic kinase efficiently and will be a useful tool to analyze its role in the pathophysiology of ALCL.     

S100A8和S100A9在冠心病中的研究进展

S100A8和S100A9作为内源性危险相关分子模式与Toll样受体4和晚期糖基化终产物受体相识别,通过参与免疫应答,改变内皮通透性并促进斑块内炎症,从而影响动脉粥样硬化的发生和发展。此外,S100A8和S100A9血浆水平升高与增加心血管事件的风险密切相关。本文综述S100A8和S100A9的研究现状及其在冠心病的预防、治疗及预后评估中的应用前景。     

S100A16基因促进3T3-L1前脂肪细胞分化

目的 研究S100A16基因在3T3-L1前脂肪细胞分化过程中的作用及机制.方法 构建过表达S100A16的慢病毒载体(PLJMI-S100A16-GFP),转染3T3-L1细胞.以Western印迹法检测S100A16正常3T3-L1细胞分化过程中S100A16的表达;采用油红O观察脂滴堆积情况;采用Western印迹和实时定量PCR方法检测前体脂肪细胞分化过程中相关基因的表达变化.免疫共沉淀方法检测S100A16是否与p53相互作用.结果 成功构建S100A16过表达3T3-L1细胞株;随着3T3-L1前脂肪细胞的分化,S100A16蛋白表达水平逐渐升高;高表达S100A16能够促进3T3-L1前脂肪细胞分化,促进甘油三酯在脂肪细胞内聚集(P＜0.01),同时上调脂肪细胞分化标志基因PPARy、CCAAT增强子结合蛋白α(C/EBP-α)、脂蛋白脂酶、脂肪细胞脂肪酸结合蛋白(aP2)及脂肪酸合成酶的表达(P＜0.05或P＜0.01);免疫共沉淀结果提示,S100A16蛋白与p53相互作用.结论 S100A16通过抑制p53活性进而促进3T3-L1前脂肪细胞的分化.     

人肺腺癌组织S100 A6蛋白表达及其临床意义

目的：探讨S100A6蛋白表达对肺腺癌患者的临床特征及其转移预后的影响。方法收集手术切除、经病理证实的肺腺癌及配对癌旁正常肺组织标本各98例,通过 Western blot 检测S100A6蛋白表达。结果肺腺癌组织 S100A6蛋白表达量显著高于癌旁正常肺组织(t=19.884,P<0.05);S100A6蛋白表达与肺腺癌患者性别、年龄无相关性(P 值均>0.05),与患者吸烟指数、肿瘤细胞分化、淋巴结转移、远处转移、临床分期均有显著相关(P 值均<0.05);Kaplan-Meier 法比较低表达组和高表达组患者的生存期,差异有统计学意义(P <0.05);COX比例风险多因素回归分析发现,肿瘤分化、TNM分期和 S100A6蛋白表达是影响肺腺癌患者预后的独立因素。结论 S100A6在肺腺癌组织显著高表达,可以作为与肿瘤发生、发展密切相关的一种标志蛋白。     

S100A4 regulates motility and invasiveness of human esophageal squamous cell carcinoma through modulating the AKT/Slug signal pathway

The involvement of S100A4 in modulating invasiveness of esophageal squamous cell carcinoma (ESCC) cell lines was explored. It was shown that S100A4 expression is positively correlated with the degree of invasiveness in human ESCC cells. The S100A4‐rich EC‐1 cells displayed higher migratory and invasive cell behavior while ET‐1 cells with low S100A4 expression levels displayed lower migratory and invasive cell behavior. S100A4 silencing by small interfering (siRNA) in EC‐1 cells induced E‐cadherin expression, and overexpression of S100A4 in a lowly invasive TE‐1 cells suppressed E‐cadherin expression. It is suggested that S100A4 silencing inhibit invasion via E‐cadherin upregulation, and overexpression of S100A4 promote invasion via E‐cadherin downregulation in ESCC cells. Compared with the vector‐transfected cells, S100A4 silencing in EC‐1 cells showed reduced ability of migration and invasiveness, and overexpression of S100A4 in TE‐1 cells showed increased ability of migration and invasiveness via wound‐healing and Transwell assay, and pseudometastatic model assay. Furthermore, re‐expression of S100A4 could increase the invasive phenotypes in S100A4 siRNA transfected EC‐1 cells, and S100A4 silencing could decrease the invasive phenotypes in S100A4 circular DNA (cDNA) transfected TE‐1 cells. It was found that Slug is downregulated in S100A4 siRNA transfected EC‐1 cells, and Slug is upregulated in S100A4 cDNA transfected TE‐1 cells. It was also discovered S100A4 cDNA induced protein kinase B (AKT) phosphorylation at Serine‐473(phospho‐AKT [p‐AKT]) levels, followed by the Slug upregulation, and S100A4 siRNA decreases the phospho‐AKT levels, followed by the Slug downregulation. The data suggested that S100A4 could regulate migratory and invasive behavior of human ESCC cells through modulating AKT/Slug pathway.     

The tyrosine kinase NPM-ALK, associated with anaplastic large cell lymphoma, binds the intracellular domain of the surface receptor CD30 but is not activated by CD30 stimulation

The heterogenous group of anaplastic large cell lymphomas (ALCLs) is characterized by expression of the Ki-1/CD30 antigen, a member of the tumor necrosis factor receptor superfamily. About 40 to 50% of cases diagnosed as ALCL contain a specific chromosomal rearrangement, t(2;5)(p23;q35), resulting in expression of the chimeric tyrosine kinase NPM-ALK. As NPM-ALK-positive lymphomas define a distinct subtype within the group of ALCL, the chimeric protein might be responsible for certain pathogenetic and clinicopathologic characteristics. To better elucidate the function of NPM-ALK, we investigated a possible mechanism for regulation of its activity. We demonstrate that NPM-ALK specifically binds to the intracellular domain of the cytokine receptor CD30. In vitro binding assays revealed that the ALK portion of NPM-ALK mediates interaction of the two proteins. Stimulation of the CD30 receptor by cross-linking with immobilized anti-CD30 antibody results in complete growth inhibition of Karpas 299, an NPM-ALK-positive ALCL cell line, but does not alter proliferation of HDLM-2, a Hodgkin's lymphoma-derived cell line lacking t(2;5). Western blot analysis of coimmunoprecipitated CD30 and NPM-ALK proteins from stimulated Karpas 299 cells showed that the interaction of the proteins is not modified by stimulation. Activation of CD30 neither enhanced NPM-ALK activity measured by autophosphorylation of the chimeric tyrosine kinase nor phosphorylation of phospholipase C-gamma, an NPM-ALK substrate. We conclude that NPM-ALK is not stimulated by CD30 activation, but exists as a constitutively hyperactivated protein. Interaction with CD30 may extend the subcellular localization of NPM-ALK to the microenvironment of membrane-associated proteins.     

Pediatric ALK+ anaplastic large cell lymphoma with t(3;8)(q26.2;q24) translocation and c-myc rearrangement terminating in a leukemic phase

Pediatric ALK-positive anaplastic large cell lymphoma (ALK+ ALCL) is usually associated with a favorable prognosis. ALK+ ALCL associated with a leukemic phase is uncommon, but has been associated with an aggressive clinical course and unfavorable prognosis. Overexpression of c-myc has been shown to be a consistent finding in ALK+, but not ALK-negative ALCL (ALK- ALCL), and the c-myc gene is considered a downstream target of deregulated ALK signaling. We describe a pediatric ALK+ ALCL with a leukemic phase at relapse. Similar to other rare cases described in the literature, it followed an aggressive clinical course despite multiple regimens of chemotherapy and bone marrow transplantation. Lymphoma cells showed aberrant ALK expression and c-myc overexpression. In addition to the characteristic t(2;5)(p23;q35) translocation, a t(3;8)(q26.2;q24) translocation was also present, and c-myc gene rearrangement was confirmed by FISH analysis. The findings in this case demonstrate the association of peripheral blood leukemic involvement and aggressive clinical course, and suggest that other factors, such as c-myc rearrangement, may be responsible for the aggressive clinical behavior in ALK+ ALCL.     

S100A8 enhances IL-1β production from nasal epithelial cells in eosinophilic chronic rhinosinusitis

BackgroundChronic rhinosinusitis is classified into eosinophilic chronic rhinosinusitis (ECRS) and non-eosinophilic chronic rhinosinusitis (NECRS). ECRS is a refractory allergic disease involving a variety of immune and epithelial cells. S100A8 is a damage-associated molecular pattern that is closely related to allergic inflammation. However, the pathological implications of S100A8 in ECRS have not been clarified.MethodsWe evaluated the role of S100A8 in the pathogenesis of ECRS. Gene expression profiles of nasal polyps obtained from patients with ECRS or NECRS were evaluated using RNA sequencing.ResultsS100A8 was identified as a significantly upregulated gene in nasal polyps associated with ECRS. Immunohistochemistry consistently revealed intense S100A8 staining in nasal polyps from patients with ECRS. Human nasal epithelial cells expressed the receptor for advanced glycation end products and Toll-like receptor 4. Recombinant S100A8 protein induced interleukin-1β secretion in human nasal epithelial cells.ConclusionsOur data demonstrate that S100A8 results in production of interleukin-1β in the nasal epithelium, which may be involved in the pathogenesis of ECRS.     

Prognostic significance of S100A4 and vascular endothelial growth factor expression in pancreatic cancer

AIM： To investigate the expression of vascular endothelial growth factor （VEGF） and calcium-binding protein S100A4 in pancreatic cancer and their relationship to the clinicopathological parameters and prognosis of pancreatic cancer. METHODS： Expression status of VEGF and S100A4 was examined in 62 surgical specimens of primary pancreatic cancer by immunohistochemistry. Correlation between the expression of VEGF and S100A4 and clinicopathological parameters was analyzed. RESULTS： Thirty-eight of 62 （61.3%） specimens of primary pancreatic cancer were positive for S100A4. Thirty-seven （59.7%） specimens showed positive expression of VEGF. The positive correlation between S100A4 and VEGF expression was significant in cancer tissues （P 〈 0.001）. S100A4 expression was significantly correlated with tumor size, TNM stage and poorer prognosis. VEGF expression had a significant correlation with poorer prognosis. The prognosis of 17 S100A4- and VEGF-negative cancer patients was significantly better than that of other patients （P 〈 0.05）. Distant metastasis （P = 0.001）, S100A4- （P = 0.008） and VEGF-positive expression （P = 0.016） were significantly independent prognostic predictors （P 〈 0.05）. CONCLUSION： Over-expression of S100A4 and VEGF plays an important role in the development of pancreatic cancer. Combined examination of the two molecules might be useful in evaluating the outcome of patients with pancreatic cancer.     

胰腺癌S100A4癌基因及ppENK抑癌基因的甲基化状态

目的 分析胰腺癌及胰腺癌细胞株ppENK、S100A4基因的甲基化状态.方法 收集31例胰腺癌及相应癌旁组织、5株胰腺癌细胞株及1例正常胰腺组织.采用MSP方法分析ppENK基因甲基化状态,采用COBRA方法分析S100A4基因甲基化状态,RT-PCR检测ppENK和S100A4 mRNA,免疫组化法检测S100A4蛋白表达,并与胰腺癌临床参数进行相关性分析.结果 正常胰腺组织ppENK基因无甲基化,S100A4基因高甲基化.31例胰腺癌组织中ppENK基因甲基化率为90.3%,与临床参数无相关性;S100A4基因低甲基化率为71.0%,仅与外周血CA19-9水平呈相关性(P=0.011,OR=0.05).S100A4蛋白在胰腺癌组织中表达率为87.1%,该表达与S100A4基因低甲基化相关(P=0.017),同时与肿瘤的分化程度有关,肿瘤分化程度越低,表达阳性率越高.S100A4基因低甲基化与ppENK基因甲基化相关(P=0.019).5株胰腺癌细胞株ppENK基因均甲基化,ppENK mRNA不表达;S100A4基因均低甲基化,S100A4 mRNA均高表达.结论 胰腺癌组织ppENK基因为高甲基化状态,而S100A4基因为低甲基化状态.