喉癌中microRNA-24对S100A8基因3'非翻译区的作用

目的 分析喉癌中S100A8基因3'UTR与其靶向miRNA的调控作用.方法 构建S100A8基因3'UTR野生型和突变型荧光素酶载体,应用MiRanda和RNA22软件预测S100A8基因3'UTR靶向miRNA,共转构建的荧光素酶载体和靶miRNA前体于喉癌Hep2细胞中,双荧光素酶检测系统测定荧光素酶活性.结果 ...     

S100A8/S100A9在骨关节炎疾病中的研究进展

S100A8(钙粒蛋白A)和S100A9(钙粒蛋白B)是钙结合蛋白S100家族的重要成员,作为重要的促炎介质,参与多种慢性炎症性疾病的病理进程,在炎症反应和机体固有免疫过程中发挥重要作用.最近有研究表明S100A8和S100A9蛋白在骨关节炎炎症的病理发展中发挥关键作用,针对S100A8和S100A9的干预有望成为骨关节炎临床治疗的潜在靶点.     

外源性S100A8在体外抑制HeLa细胞的增殖和侵袭

目的 探讨外源性S100A8对宫颈癌细胞系HeLa的增殖、凋亡、克隆形成及迁移和侵袭的影响.方法 MTT法检测细胞增殖活性;Hoechst染色检测细胞凋亡;流式细胞技术检测细胞周期;平板集落形成实验检测细胞集落形成;划痕和Transwell侵袭实验分别检测细胞的迁移和侵袭.结果细胞培养3d时,浓度为100、300和10...     

外源性S100A8在体外抑制HeLa细胞的增殖和侵袭

目的探讨外源性S100A8对宫颈癌细胞系HeLa的增殖、凋亡、克隆形成及迁移和侵袭的影响。方法 MTT法检测细胞增殖活性;Hoechst染色检测细胞凋亡;流式细胞技术检测细胞周期;平板集落形成实验检测细胞集落形成;划痕和Transwell侵袭实验分别检测细胞的迁移和侵袭。结果细胞培养3 d时,浓度为100、300和1 000 mg/L的GST-hS100A8组的A值较GST组减少13.64%、19.29%和25.06%(P<0.05);在浓度为100 mg/L时,GST-hS100A8组呈时间依赖性减少(P<0.05);GST-hS100A8组在第3天时细胞凋亡率较GST组增加5.18倍(P<0.05),同时,出现峰值为(19.9±0.76)%的凋亡峰,而对照组没有凋亡峰;GST-hS100A8组的集落形成率较GST组减少30.2%(P<0.005);划痕愈合率较GST组降低30.1%(P<0.05);穿膜细胞数减少48.9%(P<0.05)。结论外源性S100A8可能具有抑制宫颈癌的作用。     

D-半乳糖诱导的衰老大鼠心脏S100A8/A9及相关炎症通路基因表达的研究

目的：探讨D-半乳糖（D-Gal）诱导的衰老大鼠模型心脏差异基因表达的变化,并对其中S100钙结合蛋白A8/A9(S100A8/A9)及其下游p38丝裂原活化蛋白激酶（MAPK）/核因子κB(NF-κB)通路关键因子进行验证。方法：（1）将8～10周龄SD大鼠随机分为正常对照（control）组和D-Gal组,每组6只。D-Gal组大鼠每天颈背部皮下注射D-Gal(150 mg/kg),control组大鼠注射相同体积的生理盐水。Morris水迷宫检测学习记忆能力;测定血清超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和丙二醛（MDA）水平;超声心动图测定心功能变化;进行心脏HE、Masson、衰老相关β-半乳糖苷酶（SA-β-Gal）和二氢乙啶染色,并用透射电镜观察线粒体超微结构。转录组测序分析大鼠心脏基因表达差异,qRT-PCR和Western blot检测大鼠心肌组织衰老标志基因、S100A8/A9、p38 MAPK和NF-κB通路相关因子的mRNA和蛋白表达。（2）利用D-Gal(10 g/L)诱导建立H9C2心肌细胞衰老模型,并用S100A8/A9抑制剂帕奎莫德（paquin...     

非小细胞肺癌中S100A2、S100A4及S100P表达及意义

目的研究钙结合蛋白S100家族中S100A2、S100A4及S100P基因在非小细胞肺癌(non-small cell lung cancer,NSCLC)中的表达,阐明其与肺癌发生及转移的关系。方法以12例正常肺组织为对照,采用半定量RT-PCR技术检测17例腺癌和12例鳞癌及其癌旁组织中S100A2、S100A4及S100P mRNA的表达水平。结果(1)S100A2、S100A4及S100P mRNA在NSCLC中的表达量均高于癌旁和正常组织。(2)肺腺癌中三者mRNA表达量均高于癌旁和正常组织;肺鳞癌中S100A4和S100P mRNA的表达量高于正常组织。(3)根据不同的临床分期,S100A2、S100A4 mRNA在Ⅱ期与Ⅲ期中的表达量均高于Ⅰ期;S100P mRNA在Ⅲ期中的表达量高于Ⅰ期。(4)根据有无淋巴结转移,三者mRNA在有淋巴结转移癌组织中的表达量均高于无淋巴结转移的癌组织。(5)根据有无静脉癌栓,S100A4 mRNA在有静脉癌栓的癌组织中的表达量高于无静脉癌栓的癌组织。结论S100A4、S100A6、S100P在NSCLC中表达增加,尤其是在有淋巴结转移及TMN分期越高的癌组织中表...     

siRNA干扰沉默S100A4基因表达对喉癌Hep2细胞周期和侵袭的影响

目的应用RNA干扰技术下调喉癌Hep2细胞中S100A4基因的表达,探讨其对Hep2细胞周期和细胞侵袭力的影响。方法脂质体法转染S100A4siRNA至Hep2细胞。Real-time PCR和Western blot验证S100A4基因mRNA和蛋白水平的表达,流式细胞术和transwell实验分别检测下调S100A4表达对Hep2细胞周期和侵袭能力的影响。结果 S100A4 mRNA和蛋白表达水平明显降低,细胞增殖指数显著降低,并且处于G1期细胞数明显增多,细胞侵袭能力显著降低。结论 S100A4siRNA转染喉癌Hep2细胞能有效下调S100A4基因的表达,从而影响Hep2的细胞周期和抑制细胞的侵袭能力。     

鼠抗人S100A9 天然蛋白单克隆抗体的制备与鉴定

目的:探索以白细胞为免疫原的鼠抗人白细胞天然蛋白单克隆抗体(monoclonal antibodies,mAbs)制备与克隆筛选鉴定方法,制备鼠抗人S100A9 天然蛋白单克隆抗体,并鉴定抗体性质。方法:用健康人外周血白细胞免疫小鼠,利用B淋巴细胞杂交瘤技术制备mAbs,通过免疫细胞化学法对杂交瘤细胞进行非特异性阳性筛选,有限稀释法进行亚克隆,应用免疫沉淀鄄质谱法进行mAb 特异性鉴定,通过Western blot 进行细胞株筛选,小鼠体内诱生腹水法制备单抗,亲和层析法纯化,ELISA 间接法测定效价及亲和力,Western blot 进行特异性鉴定及交叉反应性分析,免疫组化染色人乳腺癌石蜡切片。结果:获得免疫细胞化学检测阳性多克隆细胞35 孔,分泌鼠抗人S100A9 蛋白单克隆抗体细胞株11 株,优选1 株制备腹水并纯化鉴定抗体,抗体效价为1 3.18 105 ,亚类为IgG1,轻链为kappa 链,抗体纯度达95%以上,亲和力常数3.54 108 L/ mol,与S100A8的交叉反应率为0.12%,与S100A12 和S100A13 几乎无交叉反应,组化染色识别人乳腺癌组织中的S100A9 蛋白。结论:成功制备鼠抗人S100A9 天然蛋白单克隆抗体,该单抗具有高效价、较高亲和力和高特异性,能为其应用于免疫组化检测S100A9蛋白的表达提供依据。     

钙结合蛋白S100A2与肿瘤

钙结合蛋白家族成员其结构相似，都具有与钙离子结合的区域，但功能不尽相同，特别是与肿瘤的关系研究显示出不同的功能作用。S100A2是唯一的与肿瘤发生发展呈负相关的蛋白。本文将对钙结合蛋白S100A2基因的结构、功能、定位、组织分布及其与肿瘤的关系等加以回顾，并对其在肿瘤基因治疗中的作用进行阐述。     

S100A8作为非小细胞肺癌免疫检查点抑制剂治疗疗效预测指标的研究

目的探讨S100A8作为非小细胞肺癌(NSCLC)免疫检查点抑制剂(ICIs)治疗疗效预测指标的可行性与潜在机制。方法采用在线数据库,分析NSCLC肿瘤组织和正常肺组织中S100A8 mRNA、蛋白的表达情况以及S100A8 mRNA水平与NSCLC患者生存情况的关系。采用生物信息学分析法分析S100A8与肿瘤组织中免疫抑制性细胞、免疫抑制性分子、缺氧和血管生成的相关性。采用Kaplan-Meier对数秩检验分析血浆S100A8水平与接受ICIs治疗的NSCLC患者无进展生存期(PFS)的关系。结果肿瘤组织中,S100A8 mRNA和蛋白水平均高于正常肺组织(P=0.006 1)。S100A8 mRNA水平升高提示NSCLC患者生存预后差(P<0.000 1)。S100A8 mRNA水平越高,髓源性免疫抑制细胞(MDSC)(P<0.000 1)、调节性T细胞(Treg)(P<0.000 1)、M2型巨噬细胞(P<0.000 1)浸润的越多,IL-10生成的越多(P<0.000 1)且缺氧(P<0.000 1)和血管生成(P=0.007 4)越严重。生存曲线显示血浆S100A8水平升高提示接受ICIs治疗的NSCLC患者无进展生存期受损(P<0.05),且S100A8是PFS的独立影响因素(P<0.05)。结论 S100A8升高提示NSCLC患者处于免疫抑制状态,且接受ICIs治疗时PFS减短,S100A8具有作为NSCLC患者ICIs治疗疗效预测指标的可能性。     

S100A8, S100A9 and the S100A8/A9 heterodimer complex specifically bind to human endothelial cells: identification and characterization of ligands for the myeloid-related proteins S100A9 and S100A8/A9 on human dermal microvascular endothelial cell line-1 cells

The natural ligands of the S100 EF hand proteins S100A8 and A9 [myeloid-related proteins 8 and 14] have long been searched for in order to further the understanding of the role of the S100A8/A9-expressing monocyte subpopulation in progressing inflammatory processes. We demonstrate that S100A8, S100A9 and the S100A8/A9 heterodimeric complex bind to human dermal microvascular endothelial cell line (HMEC)-1 with an increasing binding capacity progressing from S100A8 < or = S100A9 < or = S100A8/A9. Similar results were obtained in the apolipoprotein E knockout mouse model, where preferably recombinant S100A9 but no S100A8 bound to the endothelium of the aorta ascendens. The binding of the S100A8/A9 heterodimer complex to activated HMEC-1 is specific as demonstrated by a dose-responding and satiable binding curve and the competition of FITC-labeled versus unlabeled protein. The protein character of the binding site was proven by treatment with trypsin. S100A8/A9 binding to HMEC-1 is inducible by lipopolysaccharide and tumor necrosis factor-alpha, and in the presence of calcium. A 163-kDa protein was isolated from a cell lysate of activated HMEC-1 cells using an affinity-chromatography protocol. The endothelial cell-associated ligand proteins isolated by the use of the S100A9 monomer and the S100A8/A9 dimer were subjected to mass spectrometry for protein identification. Clearly, alpha(2)-macroglobulin was identified as a binding partner for the S100A9 monomer, whereas no protein could be identified from the database for the ligand of the S100A8/A9 dimer.     

Expression of S100 proteins in normal human tissues and common cancers using tissue microarrays: S100A6, S100A8, S100A9 and S100A11 are all overexpressed in common cancers

AIMS: To survey the expression of members of the S100 family of calcium-binding proteins in normal human tissues and common cancers using tissue microarrays. S100A6, S100A8, S100A9 and S100A11 have all been suggested to have potential roles in carcinogenesis and tumour progression but their expression has not been described in a wide range of human tissues and tumours. METHODS AND RESULTS: A custom-made tissue array, containing 291 tissue cores representing 28 tissue types and 21 tumour types, was used to produce sections that were immunostained for S100A2, S100A6, S100A8, S100A9, S100A11, calbindin 1, calbindin 2, S100B and parvalbumin. S100A6, S100A8 and S100A9 were expressed in 32%, 12% and 28% of breast cancers, respectively. There was a translocation of S100A11 expression from exclusively nuclear in normal tissues to cytoplasmic and nuclear in all common cancers. CONCLUSIONS: S100A6, S100A8, S100A9 and S100A11 are all expressed in common cancers, especially breast cancer. In addition, S100A11 undergoes a nucleocytoplasmic translocation which may have a direct influence on the proliferation of the cancer cells.     

Expression of S100A2 and S100A6 in thyroid carcinomas

AIMS: S100 calcium-binding proteins are known to play multiple roles in carcinoma development. In this study, we focused on two kinds of these proteins, S100A2 and S100A6, and investigated their expression in thyroid neoplasms. METHODS AND RESULTS: We investigated S100A2 and S100A6 expression in 141 thyroid neoplasms by immunohistochemistry. S100A2 was not expressed in normal follicles or follicular tumours, with one exception. Although 89.5% of papillary carcinoma were positive for S100A2, the expression was heterogeneous except in two cases. In anaplastic carcinoma, 78.5% of cases expressed S100A2 diffusely, while the remaining cases were negative. In normal follicles, S100A6 expression was always low, while 8.3% of follicular adenomas and 39.5% of follicular carcinomas showed increased expression. In papillary carcinomas, S100A6 expression was increased in 75% of cases, but in anaplastic carcinomas it was decreased, with only 14.3% showing high expression. CONCLUSIONS: The expression patterns of S100A2 and S100A6 in thyroid neoplasms are unique compared with those of other carcinomas, suggesting that: (i) S100A2 and S100A6 contribute to certain events in papillary carcinoma progression, and (ii) S100A2 expression is one of the biological characteristics of anaplastic carcinoma.     

S100A8, S100A9 and S100A12 activate airway epithelial cells to produce MUC5AC via extracellular signal‐regulated kinase and nuclear factor‐κB pathways

Airway mucus hyperproduction is a common feature of chronic airway diseases such as severe asthma, chronic obstructive pulmonary disease and cystic fibrosis, which are closely associated with neutrophilic airway inflammation. S100A8, S100A9 and S100A12 are highly abundant proteins released by neutrophils and have been identified as important biomarkers in many inflammatory diseases. Herein, we report a new role for S100A8, S100A9 and S100A12 for producing MUC5AC, a major mucin protein in the respiratory tract. All three S100 proteins induced MUC5AC mRNA and the protein in normal human bronchial epithelial cells as well as NCI‐H292 lung carcinoma cells in a dose‐dependent manner. A Toll‐like receptor 4 (TLR4) inhibitor almost completely abolished MUC5AC expression by all three S100 proteins, while neutralization of the receptor for advanced glycation end‐products (RAGE) inhibited only S100A12‐mediated production of MUC5AC. The S100 protein‐mediated production of MUC5AC was inhibited by the pharmacological agents that block prominent signalling molecules for MUC5AC expression, such as mitogen‐activated protein kinases, nuclear factor‐κB (NF‐κB) and epidermal growth factor receptor. S100A8, S100A9 and S100A12 equally elicited both phosphorylation of extracellular signal‐regulated kinase (ERK) and nuclear translocation of NF‐κB/degradation of cytosolic IκB with similar kinetics through TLR4. In contrast, S100A12 preferentially activated the ERK pathway rather than the NF‐κB pathway through RAGE. Collectively, these data reveal the capacity of these three S100 proteins to induce MUC5AC production in airway epithelial cells, suggesting that they all serve as key mediators linking neutrophil‐dominant airway inflammation to mucin hyperproduction.