钙结合蛋白S100A12在急性感染性疾病中应用价值的研究进展

近年来,急性感染性疾病的新型生物学标志物不断被发现,其中钙结合蛋白S100A12主要在人外周血细胞中表达,是一种炎性反应蛋白,也是钙结合蛋白S100家族的重要成员。钙结合蛋白S100A12检测方法简单、方便、快捷,在急性感染性疾病尤其是败血症、脓毒症、细菌性肺炎的鉴别诊断、严重程度评估、治疗等方面具有较高的临床应用价值,可作为急性感染性疾病的新型生物学标志物。本文主要综述了钙结合蛋白S100A12在急性感染性疾病中的应用价值,以期为提高临床急性感染性疾病诊治水平提供参考。     

S100A9蛋白的生物学特性与临床意义

S100A9蛋白属钙结合蛋白S100蛋白家族成员,通常与S100A8通过化学结合形成异二聚体,其主要功能是抑制酪蛋白激酶Ⅰ和Ⅱ的活性。近年来研究发现在多种疾病状态,包括肿瘤组织中有S100A9高表达,这提示S100A9有可能作为临床诊断的新指标。     

钙结合蛋白S100A12在动脉粥样硬化中的作用

钙结合蛋白S100A12作为具有EF手型结构的S100蛋白家族中的较新一员,有着其独特的结构与功能。人S100A12主要由中性粒细胞表达和分泌,也可被单核细胞及巨噬细胞诱导表达,其通过Ca2+、Zn2+等调节构象变化,结合特异性配体在体内参与多种生理及病理过程。S100A12作为单核细胞趋化剂,活化肥大细胞,在体内形成嗜中性粒细胞和巨噬细胞聚集。因此,S100A12可能对动脉粥样硬化的形成、进展及预后起着非常重要的作用。     

血清S100A8、S100A9水平与小儿难治性肺炎支原体肺炎病情严重程度及预后的关系

目的 探讨血清S100钙结合蛋白A8(S100A8)、S100钙结合蛋白A9(S100A9)水平与小儿难治性肺炎支原体肺炎（RMPP）病情严重程度及预后的关系。方法 选取70例RMPP患儿为RMPP组、80例普通肺炎支原体肺炎患儿为GMPP组、50例健康体检儿童为对照组。根据病情严重程度将RMPP患儿分为轻症组（n=36）、重症组（n=34）,根据预后将RMPP患儿分为预后良好组（n=43）和预后不良组（n=27）。用酶联免疫吸附实验检测血清S100A8、S100A9并比较各组二者水平变化。分析S100A8、S100A9水平与RMPP患儿病情严重程度的关系;Logistic回归分析RMPP患儿预后不良的危险因素;绘制受试者工作特征曲线,用曲线下面积评价血清S100A8、S100A9水平对RMPP患儿预后的预测效能。结果 RMPP组血清S100A8、S100A9水平均高于GMPP组和对照组（P均<0.05）,GMPP组血清S100A8、S100A9水平均高于对照组（P均<0.05）。治疗前及治疗后7 d,重症组血清S100A8、S100A9水平均高于轻症组（P均<0.0...     

钙结合蛋白S100A4与消化系肿瘤研究进展

S100A4基因编码的钙离子结合调节蛋白-S100A4蛋白,通过与钙离子结合,在许多肿瘤发展、转移过程中发挥重要作用.目前研究认为其与肿瘤的浸润和转移密切相关.本文就S100A4生物学特性及其在消化系肿瘤中作用及可能机制的研究进展作一综述.     

shRNA干扰S100A13基因对人甲状腺癌细胞释放成纤维细胞生长因子1的影响

目的 探讨S100A13基因沉默对应激诱导人甲状腺癌细胞(TT细胞)成纤维细胞生长因子1(FGF-1)释放的影响.方法 将S100A13-shRNA pENTRTM/U6入门质粒转染TT细胞,应用实时PCR、间接免疫荧光法及Western印迹方法检测细胞内S100A13基因及蛋白表达的抑制效率,应用间接免疫荧光,RT-PCR及ELISA检测无血清处理TT细胞S100A13沉默后FGF-1释放变化.结果 S100A13-shRNApENTRTM/U6入门载体转染TT细胞后能够抑制S100A13基因及蛋白表达,抑制效率约为80%.间接免疫荧光法显示FGF-1主要位于细胞浆和细胞核,无血清培养6 h后胞浆内FGF-1基本消失.RT-PCR和ELISA结果均显示,S100A13沉默能降低无血清处理TT细胞培养上清中FGF-1的浓度(P＜0.05).结论 S100A13-shRNA pENTRTM/U6入门载体能有效、特异性地抑制S100A13基因及蛋白表达.S100A13基因的抑制可以减弱FGF-1从细胞内释放到细胞外的过程.     

S100A6、S100B对非小细胞肺癌的变化及与预后的意义

目的分析S100A6、S100B对非小细胞肺癌(NSCLC)患者中的变化及与预后的意义。 方法选取2016年10月至2018年9月我院收治的76例NSCLC患者作为对象,治疗前及治疗6个月后检测S100A6、S100B蛋白表达水平,分析S100A6、S100B对NSCLC预后的预测意义。 结果76例NSCLC患者的3年生存者11例(14.47%),死亡者65例,中位生存时间为15.5个月。死亡者治疗前后S100A6、S100B蛋白水平高于生存者(P<0.05)。死亡者TNM分期Ⅲb~Ⅳa期及出现骨转移所高于生存者,KPS评分低于生存者(P<0.05)。TNM分期Ⅲb~Ⅳa期及出现骨转移、KPS评分低、S100A6、S100B蛋白表达水平高为NSCLC预后的影响因素(P<0.05)。ROC结果,S100A6、S100B蛋白水平及联合对NSCLC预后预测的AUC分别为0.763(95%CI：0.649~0.876)、0.801(95%CI：0.694~0.907)、0.849(95%CI：0.731~0.952)。 结论S100A6、S100B蛋白表达水平对NSCLC患者预后具有临床意义。     

钙结合蛋白S100A12与冠状动脉粥样斑块稳定性的相关性

目的 探讨冠心病患者血清钙结合蛋白S100A12的表达及其与冠心病的相关性,评价其作为预测斑块稳定性的外周血生物学指标的临床意义.方法 选择依据临床症状及冠脉造影结果冠心病诊断明确的患者91例,其中稳定型心绞痛（SAP组）18例、不稳定型心绞痛（UAP组） 37例、急性心肌梗死（AMI组）（包括ST段抬高型心梗和非ST段抬高型心梗） 36例.对照组为冠脉造影正常或微小病变＜50%排除冠心病的患者34例.观察血清S100A12水平在各组间的变化.通过介入手术模拟斑块破裂,比较术前、术后S100A12水平变化,评价其预测斑块稳定性的临床意义.结果 ①冠心病患者（SAP组、UAP组、AMI组）血清S100A12水平显著高于对照组（P＜0.05）;与SAP组比较,UA组及AMI组血清S100A12水平显著升高,且差异有统计学意义（P＜0.05）;UAP组与AMI组比较血清S100A12水平差异无统计学意义（P＞0.05）.②根据是否行支架置入术分为单纯行选择性冠脉造影组（CAG组）与选择性冠脉造影＋内支架置入术组（PCI组）.在CAG组中,血清S100A12术后与术前即刻相比差异无统计学意义（P＝0.064）;在PCI组中,术后血清S100A12水平与术前即刻相比显著升高,差异有统计学意义（P＜0.01）.③入院至手术前住院期间药物对血清S100A12水平的影响：所有对象入院时与术前即刻血清S100A12水平相比差异无统计学意义（P＞0.05）.结论 钙结合蛋白S100A12可能参与动脉粥样硬化的形成,并且可能成为外周血中预测斑块稳定性的生物学指标.     

S100A2基因在结直肠癌组织中的表达、突变及杂合性缺失

目的:探讨S100A2基因在结直肠癌发生及演进过程中的作用.方法:对66例结直肠癌组织标本进行DNA、mRNA及蛋白提取.采用PCR-SSCP和微卫星结合PCR-变性聚丙烯酰胺凝胶电泳技术检测S100A2基因DNA突变及缺失:分别以RT-PCR技术、Western blot检测S100A2 mRNA及蛋白表达水平.结果:66例结直肠癌组织标本中,未发现S100A2基因存在杂合性缺失(LOH)及第二外显子和第三外显子突变:S100A2 mRNA及蛋白表达水平在结直肠癌中明显低于正常结直肠组织(0.499±0.307 vs 1.187±0.264;0.542±0.193 vs 1.301±0.233.均P＜0.05).结论:S100A2蛋白表达下调可能与结直肠癌的发生发展有关.S100A2基因在结直肠癌中表达下调可能不是杂合性缺失或突变所致.     

血清S100A9、S100A2在NSCLC患者中的变化及预后的意义

目的分析血清S100钙结合蛋白A9(S100A9)、S100钙结合蛋白A2(S100A2)在非小细胞肺癌(NSCLC)患者中的变化及预后的意义。 方法选取2019年1月至2020年10月我院收治的46例NSCLC患者为对象，选择同期在我院进行健康体检的32例为对照组。对比两组血清S100A9、S100A2水平。统计NSCLC患者预后，依据预后情况分为生存者和死亡者。Logistic多因素回归分析分析影响NSCLC患者预后的因素。绘制工作特征曲线(ROC)，以曲线下面积(AUC)分析血清S100A9、S100A2联合检测对NSCLC患者预后。 结果观察组血清S100A9、S100A2水平均高于对照组(P<0.05)。截止随访结束，病死率为26.08%。Logistic回归分析结果，临床分期为Ⅲ期、分化程度为低分化、血清S100A9及S100A2水平均为影响NSCLC患者预后的危险因素(OR＝2.824、2.790、3.401、3.102，P<0.05)。ROC结果显示，血清S100A9、S100A2预测NSCLC患者预后的最佳点分别为302.11 ng/L、7.59 μg/L，两者联合的特异度为97.92%，高于血清S100A9、S100A2, S100A9、S100A2两者联合预测NSCLC患者预后的AUC为0.899，高于S100A9、S100A2单独预测的AUC(P<0.05)。 结论血清S100A9、S100A2两者联合检测对NSCLC患者预后有意义。     

S100A6基因过表达对 A549肺腺癌细胞恶性表型特征的影响

目的：探讨 S100A6基因过表达对于肺腺癌细胞恶性表型的影响。方法构建重组pLeno-DCE-S100A6载体,慢病毒转染 A549肺腺癌细胞,应用实时荧光定量聚合酶链反应和 Western blot 鉴定 S100A6基因和蛋白表达;采用四甲基偶氮唑蓝法、Transwell 和流式细胞仪分别检测细胞增殖、侵袭、细胞周期及凋亡等恶性表型特征。结果转染 S100A6基因的 A549肺腺癌细胞 S100A6 mRNA 和蛋白的表达较空载体对照组和阴性对照组均有明显的上调(P 值均<0.05);细胞增殖和侵袭能力较空载体对照组和阴性对照组增加(P 值均<0.05);细胞周期比例在 G0/G1期与空载体对照组和阴性对照组比较差异无统计学意义(P 值均>0.05),在 S 期显著高于其他两组(P 值均<0.05),在 G2/M 期显著低于其他两组(P 值均<0.05);平均凋亡率较空载体对照组和阴性对照组下降(P 值均<0.05)。结论肺腺癌细胞 S100A6基因过表达可增强肿瘤细胞的增殖和侵袭能力,增加DNA 合成,减弱凋亡等,与肿瘤发生、发展、侵袭及转移等生物学行为密切相关,有望作为肺腺癌诊断和治疗的分子标志物。     

下调S100A6基因对裸鼠肺腺癌移植瘤生长和凋亡的影响

目的探讨下调靶向基因S100A6对在体肺腺癌移植瘤生长和凋亡的影响。方法 18只健康Balb/c雄性裸鼠随机分为空载体对照组、阴性对照组及S100A6基因RNA干扰组三组,每组6只动物;分别应用不携带目的基因的空载质粒、未转染任何质粒以及含有S100A6基因RNA干扰慢病毒质粒的A549肺腺癌细胞接种于裸鼠皮下,构建移植瘤动物模型;观察不同组移植瘤组织生长情况;采用Real-Time PCR、免疫组织化学及Western-blot等技术检测S100A6 mRNA和蛋白的表达,TUNEL法检测细胞凋亡。结果成功构建了裸鼠肺腺癌皮下移植瘤模型;病理学特征显示,阴性对照组和空载体对照组可见到成巢的肿瘤细胞,癌细胞核大,而干扰组肿瘤细胞较稀疏,间质组织明显;接种细胞2周时,阴性对照组、空载体对照组和RNA干扰组肿瘤组织体积和质量差异具有统计学意义(P0.05);S100A6基因和蛋白表达在三组之间差异具有统计学意义(P0.05);各组裸鼠移植瘤凋亡细胞呈现不同程度的棕褐色肿瘤细胞核,三组肿瘤细胞凋亡率的差异具有统计学意义(P0.05)。结论下调肿瘤瘤组织S100A6蛋白表达,可抑制肿瘤生长,诱导细胞凋亡,为进一步开发肺腺癌分子靶点提供了新途径。     

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.     

S100A6基因沉默对胰腺癌细胞侵袭的影响

目的 探讨S100A6基因沉默对胰腺癌细胞侵袭的影响和可能机制。方法 将不同浓度(3.125、6.25、12.5 nmol/L)的靶向S100A6的小干扰RNA( S100A6-siRNA)转染人胰腺癌BxPC3细胞,分别采用荧光实时定量PCR和蛋白质印迹法检测S100A6 mRNA和蛋白的表达,采用Transwell小室检测癌细胞侵袭能力,明胶酶谱法检测基质金属蛋白酶-9(MMP-9)活性。结果 S100A6-siRNA转染组细胞的S100A6 mRNA和蛋白表达呈浓度、时间依赖性明显下调,穿膜细胞数呈浓度依赖性明显减少。12.5 nmol/L的S100A6-siRNA转染组细胞转染后48 h的S100A6 mRNA表达从对照组的(100±0.3)％降到(15.3±0.2)％(P＜0.01);S100A6蛋白的表达从(83.2±0.18)％降到(13.5±0.12)％(P＜0.01);穿膜细胞数从(44.5±2.2)个降到(7.6±1.5)个(P＜0.05)。同时,S100A6-siRNA转染组细胞的MMP-9活性明显下调。结论 抑制S100A6基因表达可抑制胰腺癌细胞侵袭转移,其机制可能与下调MMP-9活性有关。     

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。     

S100A6 as a Potential Serum Prognostic Biomarker and Therapeutic Target in Gastric Cancer

Background

Increased expression of S100A6 in many cancer tissues and its association with tumor behavior and patient prognosis were demonstrated, and there are no studies analyzing the serum levels of S100A6 in patients with gastric cancer (GC).

Aim

Serum S100A6 levels were investigated as a marker of tumor aggressiveness in patients with GC, and the S100A6 gene was examined as a potential therapeutic target in GC.

Methods

Serum S100A6 levels were detected in 103 GC patients and 72 healthy subjects by ELISA. Clinicopathological features of GC patients were analyzed in correlation to serum S100A6 levels. Two small interfering RNAs against S100A6 (siRNA1-S100A6 and siRNA2-S100A6) were generated and transfected into SGC7901 cells using pSUPER gfp-neo vector, and the effects of S100A6 knockdown on cell proliferation, invasion and apoptosis were evaluated in vitro. The effects of S100A6 silencing on tumor growth and metastasis were evaluated in vivo in a pseudo-metastatic GC nude mouse model.

Results

Serum S100A6 levels were significantly higher in GC patients than in healthy controls (P < 0.001). Serum S100A6 levels were significantly correlated with lymph node metastasis, TNM stage, perineural invasion and vascular invasion. Serum S100A6 level was an independent predictor of overall survival. SiRNA-mediated silencing of S100A6 significantly induced apoptosis and decreased proliferation, clone formation and the invasiveness of GC SGC7901 cells in vitro and significantly reduced tumor volume and number in vivo (P < 0.01).

Conclusion

Serum S100A6 level may serve as a potential prognostic biomarker in GC. Inhibition of S100A6 decreased the metastatic potential of GC cells.     

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.     

S100A4 is expressed at site of invasion in rheumatoid arthritis synovium and modulates production of matrix metalloproteinases

The metastasis-associated protein S100A4 promotes the progression of cancer by regulating the remodelling of the extracellular matrix. The expression of S100A4 in vivo is shown and the functional role of S100A4 in the pathogenesis of osteoarthritis and rheumatoid arthritisis is explored. The expression of S100A4 in rheumatoid arthritis, osteoarthritis and normal synovial tissues was determined by immunohistochemistry. The expression of matrix metalloproteinase (MMP) mRNA was measured in rheumatoid arthritis and osteoarthritis synovial fibroblasts treated and untreated with S100A4 oligomer by real-time polymerase chain reaction. Levels of released MMPs were confirmed by ELISA in cell culture supernatants. S100A4 protein was expressed in rheumatoid arthritis and osteoarthritis synovial tissues, in contrast with normal synovium. S100A4 up regulated MMP-3 mRNA in rheumatoid arthritis synovial fluid, with a peak after 6 h. This resulted in release of MMP-3 protein. MMP-1, MMP-9 and MMP-13 mRNA were also up regulated in synovial fluid, but with different kinetics. MMP-14 mRNA showed no change. Thus, S100A4 protein is expressed in synovial tissues of patients with rheumatoid arthritis and osteoarthritis in contrast with healthy people. It induces the expression and release of MMP-3 and other MMPs from synovial fluid. The data suggest that S100A4-producing cells could be involved in the pathogenesis of osteoarthritis and rheumatoid arthritis, including pannus formation and joint destruction.