重组hS100A6蛋白的原核表达及其对人骨肉瘤细胞系143B的生物学作用

目的:制备重组hS100A6蛋白,并研究其对人骨肉瘤细胞系143B的生物学作用。方法:构建pGST-HRV3C-hS100A6质粒,经转化至E.coli BL21,IPTG诱导表达融合蛋白GST-HRV3C-hS100A6,经超声破菌,谷胱甘肽-琼脂糖4B球珠纯化,GST-HRV3C酶切,再纯化,Western blot鉴定,分光光度法蛋白定量。以人骨肉瘤细胞系143B为研究对象,MTT检测细胞增殖,Hoechst检测细胞凋亡,Transwell检测细胞迁移和侵袭,Western blot检测hS100A6对β-catenin表达的影响。结果:成功制备重组hS100A6蛋白,测得该蛋白产量约为4mg/L菌液。30μg/ml重组hS100A6促进143B细胞的增殖、迁移、侵袭以及β-catenin的表达(P0.05),对143B细胞的凋亡无明显影响(P0.05)。结论:成功制备重组hS100A6蛋白,30μg/ml重组hS100A6蛋白对143B细胞有一定的促进作用。     

SAHA与顺铂联合用药对骨肉瘤细胞增殖、凋亡和迁移的影响

该文研究了组蛋白去乙酰化酶抑制剂SAHA(suberoylanilide hydroxamic acid)联合顺铂(cisplatin,DDP)对骨肉瘤细胞的作用。骨肉瘤143B细胞分别经不同浓度的SAHA、DDP、SAHA+DDP作用48 h。利用倒置显微镜观察细胞形态学变化;分别采用MTT法、流式细胞术、细胞划痕实验、克隆形成实验检测SAHA、DDP及二者联用对143B细胞活力、细胞凋亡、细胞迁移和集落形成能力的影响。利用Western blot检测143B细胞中cleaved-Caspase-8、cleaved-PARP蛋白表达水平。结果显示,SAHA+DDP组143B细胞大量坏死,抑制细胞增殖的现象明显高于单药组;中低剂量SAHA与DDP联用表现为协同作用,而较高剂量的联用表现为单纯相加作用;SAHA+DDP组较单药组细胞早期凋亡率增加、迁移率明显降低,且SAHA+DDP组的细胞集落形成能力明显降低;SAHA+DDP组能够促进cleaved-Caspase-8、cleaved-PARP的蛋白表达。以上结果表明,SAHA联合顺铂对骨肉瘤细胞143B细胞有协同增敏作用,能够明显抑制其增殖和迁移并促进其凋亡。     

甲基莲心碱促进骨肉瘤143B细胞凋亡的相关研究

目的:研究不同浓度甲基莲心碱对骨肉瘤143B细胞增值迁移的影响及其诱导凋亡的相关机制。方法:不同浓度甲基莲心碱处理骨肉瘤143B之后,CCK-8法检测甲基莲心碱对骨肉瘤143B细胞的增值抑制作用;细胞划痕实验检测不同浓度甲基莲心碱对骨肉瘤143B细胞迁移的影响;流式细胞术检测甲基莲心碱对癌细胞的凋亡率和周期分布;Western Blot检测甲基莲心碱对癌细胞相关蛋白Bcl-2、Bax的表达。结果:CCK-8显示甲基莲心碱能抑制骨肉瘤143B细胞的增值且以浓度和时间依赖的方式(P0.05);给药组(20, 40, 60μmol·L~(-1))24 h细胞迁移率分别为(62.35±4.15)%,(40.74±4.80)%,(25.10±5.52)%,较对照组(75.89±5.24)%明显降低(P0.05);流式细胞术结果显示甲基莲心碱能使骨肉瘤143B细胞周期阻滞于G0/G1期,且呈剂量依赖性诱导癌细胞凋亡(P0.05);Western Blot证明甲基莲心碱可促进癌细胞中促凋亡蛋白Bax的表达,而降低抑制凋亡蛋白Bcl-2的表达(P0.05)。结论:甲基莲心碱可能通过激活线粒体凋亡相关蛋白的表达,发挥抗骨肉瘤143B的作用。     

Caveolin-1抑制胰腺癌细胞PANC1的体内外生长和增殖

窖蛋白-1(caveolin-1)是胞膜窖(caveolae)中重要的结构和功能蛋白.Caveolin-1参与细胞的多种生命活动并与恶性肿瘤的发生相关.为探讨caveolin-1对胰腺癌细胞PANC1的体外增殖、迁移、侵袭以及裸鼠体内成瘤能力的影响,通过基因转染技术培育caveolin-1过表达细胞株PANC1/cav-1作为实验组,转染空载体细胞株PANC1/vector作为对照组,采用RT-PCR及Western blot方法检测caveolin-1的表达量,流式细胞术分析细胞周期,软琼脂细胞克隆实验检测细胞增殖能力,侵袭小室实验检测癌细胞迁移和侵袭的能力,建立裸鼠皮下种植瘤模型并检测肿瘤组织的增殖与凋亡.PANC1/cav-1中的caveolin-1表达稳定,表达量明显高于对照组细胞株和亲本细胞株(P<0.01),细胞周期检测显示大量PANC1/cav-1细胞被抑制于G0/G1期,caveolin-1抑制PANC1的增殖,迁移和侵袭能力.在裸鼠的体内实验中,caveolin-1显著抑制PANC1细胞在裸鼠体内的生长,Ki-67染色和TUNEL染色表明在PANC1细胞中过表达caveolin-1,可以抑制肿瘤增殖并诱导肿瘤凋亡.上述结果表明,caveolin-1可能通过对胰腺癌细胞周期的影响(抑制于G0/G1期),抑制胰腺癌PANC1细胞在体内外的增殖、迁移和侵袭,并导致肿瘤凋亡.     

Ad-IL-24对MG-63人骨肉瘤细胞抑癌效应的体内外实验

本研究旨在分析腺病毒携带的IL-24基因在体内外对人骨肉瘤细胞生长抑制效应及其分子机制。将Ad-IL-24重组腺病毒感染MG-63细胞,用荧光显微镜、RT-PCR法检测IL-24在MG-63细胞中的转录和表达;MTT法、流式细胞技术和Hoechst染色法检测IL-24基因的表达对MG-63细胞的生长抑制和凋亡效应;半定量RT-PCR法检测IL-24基因的表达对MG-63细胞中的bcl-2、bax、caspase-3相关基因表达的影响。用Ad-IL-24重组腺病毒在MG-63骨肉瘤荷瘤裸鼠的瘤体内进行注射治疗,观察肿瘤生长变化,15d后处死裸鼠,摘除瘤体,称瘤重。并通过免疫组化法检测Bcl-2、Bax、Caspase-3等与细胞凋亡相关因子的表达。结果表明Ad-IL-24重组腺病毒感染MG-63细胞后,能明显抑制MG-63细胞增殖,并能通过上调细胞中bax、caspase-3和下调bcl-2基因表达,诱导细胞凋亡,呈现出典型细胞凋亡形态学变化。Ad-IL-24重组腺病毒瘤内注射MG-63裸鼠荷瘤骨肉瘤移植瘤后,能显著抑制肿瘤生长,瘤重的抑制率可达52%,免疫组化结果显示Ad-IL-24重组腺病毒能明显上调与细胞...     

瘤体内注射凋亡素基因诱导人骨肉瘤S180细胞株凋亡研究

观察瘤体内注射凋亡素基因诱导人骨肉瘤S180细胞株凋亡的效果。利用脂体介导将重组质粒pCDNAVP3转染至荷瘤裸鼠肿瘤结节内。PCR和RT PCR检测结果表明 ,质粒可存在和持续表达 14d以上。抑瘤试验结果证实 ,凋亡素可抑制S180细胞的生长 ,抑瘤率达 2 7.2 %。TUNFL染色法检测结果表明 ,试验组的TUNEL阳性细胞率明显高于对照组。上述结果显示 ,所注射的凋亡素基因可诱导人骨肉瘤S180细胞株凋亡。     

电转系统介导的荧光素酶稳定表达PANC-1细胞的构建及其生物发光成像检测

在构建稳定表达荧光素酶的人胰腺癌PANC-1细胞的基础上,建立适合活体成像研究的胰腺癌动物模型,为深入研究胰腺癌发病机制打下基础。该研究运用Lonza NucleofectorTM核转染系统将携带有荧光素酶基因(firefly luciferase,luc)的质粒(GV258)稳定转染PANC-1细胞,利用嘌呤霉素筛选出能稳定表达荧光素酶的细胞株PANC-1-LUC,并应用活体成像系统对其体内外生物发光进行检测。进一步构建PANC-1-LUC裸鼠皮下移植瘤,同样方法接种慢病毒载体构建的PANC-1-luc细胞作为对照,观察成瘤率、瘤体积,检测生物发光强度。体内外生物发光检测结果均显示,光通量值与细胞数之间呈显著的直线相关(P0.01)。PANC-1-LUC细胞皮下接种裸鼠,1周成瘤率即达80%以上,瘤体积可达平均100 mm3。观察6周,裸鼠皮下肿瘤生长符合Gompertzian曲线。每周的生物发光检测结果均显示,PANC-1-LUC移植瘤的光强度与瘤体积有显著的线性关系(P0.05)。以上结果说明,该文所构建的PANC-1-LUC细胞株能够稳定、长期地表达荧光素酶,更适合构建活体成像研究的裸鼠胰腺癌模型。     

HDGF的PWWP结构域改变对肿瘤细胞体外及体内增殖的影响

目的:研究肝癌衍生生长因子(Hepatoma-derived growth factor,HDGF)PWWP结构域(PWWP domain)改变对肿瘤细胞体外及体内增殖的影响。方法:构建HDGF的PWWP结构域突变体P24A,利用慢病毒感染细胞筛选稳定细胞系。采用CCK-8法和软琼脂克隆形成实验检测细胞的增殖情况。通过裸鼠皮下成瘤实验检测移植瘤的形成情况。结果:在肝癌Hep G2和结直肠癌DLD1稳定细胞株中,CCK-8法检测结果显示突变体P24A对细胞生长的抑制作用呈时间依赖性,其OD值在24、48、72和96 h均明显低于HDGF稳定细胞株(均P0.001)。克隆形成实验结果显示P24A组克隆数目明显小于HDGF组(P0.01)。异种移植瘤动物模型则证明P24A细胞株的瘤块生长速度(P0.001,P0.01,P0.01),瘤块大小及体积(P0.01)均明显低于HDGF细胞株。结论:PWWP结构域改变可能抑制HDGF发挥促进细胞增殖的作用。     

汉黄芩素抗骨肉瘤143B细胞的实验研究

目的:观察汉黄芩素对人骨肉瘤细胞系143B增殖和凋亡的影响,并探讨其可能的作用机制。方法:采用体外培养人成骨肉瘤细胞系143B,CCK-8实验检测不同浓度汉黄芩素对骨肉瘤143B细胞增殖抑制作用;流式细胞术分析汉黄芩素对癌细胞周期分布及凋亡的影响;Western Blot检测凋亡相关蛋白Bax、Bcl-2、cleaved caspase-9和cleaved caspase-3的表达水平。结果:CCK-8结果显示汉黄芩素以时间、浓度依耐性的方式抑制骨肉瘤143B细胞的增殖;流式细胞术结果表明汉黄芩素可导致骨肉瘤细胞周期阻滞于G0/G1期并以浓度依赖的方式诱导骨肉瘤细胞凋亡;Western Blot检测结果证明,汉黄芩素可上调骨肉瘤细胞中促凋亡蛋白Bax、cleaved caspase-9、cleaved caspase-3的表达,而下调抑制凋亡蛋白Bcl-2。结论:汉黄芩素抑制骨肉瘤细胞增殖、导致细胞周期阻滞,促进其凋亡,并呈现时间和浓度依赖性,汉黄芩素激活Caspase凋亡途径及诱导细胞周期阻滞可能是其抗骨肉瘤的作用机制。     

靶向诱骗受体3的小干扰RNA抑制人结肠癌裸鼠皮下移植瘤的实验研究

目的：以脂质体为载体,探讨靶向诱骗受体3（DcR3）的小干扰RNA（siRNA）对人结肠癌SW480细胞裸鼠皮下移植瘤生长的抑制作用。方法：裸鼠背部皮下种植结肠癌SW480细胞,建立结肠癌SW480细胞裸鼠皮下移植瘤模型;针对靶标DcR3基因,利用Dharmacon公司的软件设计合成siRNA序列,通过瘤体局部多点注射脂质体与DcR3siRNA混合物,将DcR3siRNA转染到裸鼠背部皮下移植瘤内,同时设立阴性对照组和空白对照组;观察肿瘤治疗前后体积变化,评价DcR3siRNA对肿瘤的抑制作用;比较肿瘤治疗前后的细胞形态学改变;免疫组织化学及RT-PCR检测DcR3基因表达;原位细胞凋亡检测肿瘤的凋亡。结果：治疗组肿瘤体积明显小于对照组,肿瘤组织内坏死面积增大,细胞凋亡率显著增高,DcR3蛋白表达水平降低;治疗过程中裸鼠生长良好,无明显毒性反应。结论：脂质体介导的DcR3siRNA对裸鼠皮下结肠癌SW480细胞移植瘤有明显的抑制、杀伤作用,细胞凋亡是DcR3siRNA致肿瘤细胞死亡的重要形式。     

S100A6蛋白对细胞中β-catenin水平的影响及可能机制

探讨S100A6蛋白对细胞中β-catenin水平的影响及可能机制。用表达S100A6及其siRNA的重组腺病毒AdS100A6和AdsiS100A6处理人骨肉瘤细胞系143B,Western blot分析处理前后细胞中β-catenin水平的变化;再以人胚肾细胞HEK293为研究对象,通过免疫共沉淀(co-immunoprecipitation,Co-IP)联合Western blot检测S100A6与Wnt经典信号通路主要成员β-catenin、糖原合酶激酶-3β(GSK-3β)、散乱蛋白(Dishevelled,DVL)和轴蛋白(Axin)之间的相互作用。结果:1)AdS100A6作用48h后,143B细胞中β-catenin水平较实验对照组(AdGFP组)增加了64%(P<0.01),而AdsiS100A6处理的细胞中β-catenin较AdGFP组减少了20.2%(P<0.05),AdGFP组与空白组之间的差异无统计学意义;2)在S100A6抗体的沉淀物中,检测到β-catenin、GSK-3β和DVL,而无Axin检出;3)β-catenin、GSK-3β和DVL的抗体沉淀物中,均有S100A6检出,而Axin抗体的沉淀物中无S100A6检出。结论:S100A6能够上调人骨肉瘤细胞系143B中的β-catenin水平;并与β-catenin、GSK-3β和DVL之间存在直接相互作用;S100A6与这三个分子之间的直接相互作用可能抑制了β-catenin的降解,使之在胞浆内聚集,水平升高。这可能是S100A6上调β-catenin的部分机制。     

S100A9参与乙型肝炎病毒X蛋白介导的HepG2细胞增殖与迁移

目的:探讨S100A9在乙型肝炎病毒X(HBx)介导的HepG2细胞增殖及迁移中的作用。方法:用表达HBx蛋白的重组腺病毒AdHBx感染HepG2细胞后,用CCK-8实验检测细胞增殖能力及划痕愈合实验检测细胞迁移能力;在HepG2/AdHBx细胞中转染S100A9-siRNA及其对照siRNA后,检测HepG2细胞增殖及迁移能力;在HepG2/Ad HBx和对照组HepG2/AdGFP细胞中,采用Real-time PCR及Western Blot检测S100A9基因及蛋白的表达情况;在HepG2/AdHBx细胞中,加入不同剂量的NF-κB抑制剂BAY11-7082后,检测各组中S100A9的基因及蛋白表达情况。结果:HBx促进HepG2细胞的增殖与迁移; S100A9-siRNA抑制S100A9的表达后,HBx促进HepG2细胞的增殖与迁移的作用降低,HBx介导的HepG2细胞的增殖与迁移部分依赖于S100A9; S100A9基因及蛋白表达在HepG2/AdHBx中较对照组HepG2/Ad GFP显著升高,HBx可致S100A9表达增加;抑制NF-κB转录活性后,AdHBx+BAY11-7082组S100A9基因及蛋白表达较对照组显著降低,阻断NF-κB转录活性可部分抑制HBx调控的S100A9表达。结论:HBx可调控S100A9的表达且与NF-κB活化有关,S100A9参与HBx介导的HepG2细胞的增殖与迁移。     

曲古抑菌素A（TSA）对骨肉瘤细胞株143B增殖和凋亡的作用及其机制

观察曲古抑菌素A（TSA）对骨肉瘤细胞株143B增殖及凋亡的作用,并探讨其机制。TSA与p38抑制剂（SB203580,3μmol／L）及JNK抑制剂（SP600125,0．5μmol／L）单独或同时处理143B细胞,分别以MTT、台盼蓝染色、流式细胞术和JC—1（测定线粒体跨膜电位）法检,TSA对143B细胞的增殖、存活、周期以及凋亡的影响。应用RT-PCR、Westernblot检测Bax、Bcl．2、p38／JNK表达。结果显示,TsA能够以时间和剂量依赖方式抑制143B细胞增殖,使细胞周期阻滞于GdG。与G2／M期,并能诱导143B细胞凋亡,引起线粒体膜电位降低,促凋亡蛋白Bax表达上调,抑凋亡蛋白Bcl．2表达下调,同时使p38／ⅢK活化增加。p38／JNK4检测剂则能逆转TSA对Bax／Bcl．2的上调及抑制作用。研究结果揭示,TSA可以时间剂量依赖方式抑制143BN胞增殖,阻滞细胞周期,诱导细胞凋亡;其诱导细胞凋亡的机制可能与活化MAPK通路中p38和JNK的活性从而激发线粒体凋亡通路有关。     

过表达外源性CCN1对人骨肉瘤细胞系143B生长和迁移的影响

采用AdEasy系统构建带有标记基因GFP和FC的重组腺病毒AdCCN1,并感染人骨肉瘤细胞,观察外源性CCN1对肿瘤细胞生长和迁移的影响.PCR扩增FC编码序列标记的人CCN1克隆到pAdTrack-TO4中获得重组穿梭载体pAdTrack-CCN1. PmeⅠ线性化该质粒并电转到有腺病毒骨架质粒的BJ/AdEasy 1菌中,同源重组获得腺病毒质粒pAd-CCN1,PacⅠ线性化并转染293细胞,包装制备AdCCN1.卡那霉素抗性筛选、XbaⅠ与KpnⅠ酶切鉴定,荧光显微镜和Western印迹检测标记基因GFP和FC表达,鉴定获得的复制缺陷型重组腺病毒AdCCN1.AdCCN1与AdGFP分别感染人骨肉瘤细胞143B,通过MTT实验、胶原集落形成实验、划痕愈合和室移动实验,观察CCN1对143B增殖和迁移的影响.酶切鉴定表明,带有FC标记的CCN1被成功克隆到穿梭载体pAdTrack中.卡那霉素抗性筛选并酶切鉴定,获得重组腺病毒质粒pAd-CCN1.Western 印迹和荧光显微镜观察,确定与CCN1共表达的FC和GFP表达.AdCCN1感染骨肉瘤细胞, 与对照比较细胞数量增多,集落形成增加,细胞划痕逐渐愈合,迁移至膜上的细胞数量明显增加.应用AdEasy系统高效快速构建了带有标记GFP和FC的重组腺病毒AdCCN1,为监控腺病毒转基因的效果提供了便捷的工具.AdCCN1感染骨肉瘤细胞后可促进细胞增殖和迁移,提示CCN1在肿瘤发生和转移中可能起到重要作用.     

Resistance of SKW6 cell to apoptosis induction with anti-Fas antibody upon transduction of a reverse fragment to a cDNA encoding human 6A8 α-mannosidase

The effect of transduction with a reverse fragment to a cDNA encoding human 6A8 α-mannosidase on apoptosis induction of human B cell line SKW6 by anti-Fas antibody was tested. Apoptosis-inducer of anti-Fas monoclonal antibody was used to induce apoptosis in SKW6 cells. Giemsa’s staining, Annexin-V-FLUOS staining and DNA ladder test were used to determine the events of apoptosis. Indirect immunofluorescent staining with anti-Fas antibody was performed to detect the surface Fas expression. In a time-course test of 12, 24 and 36 h for apoptosis induction by anti-Fas antibody, DNA ladder was observed in the wild-type SKW6 cells in a time-dependent fashion. Mock transduction had no effect on DNA ladder production. However, no DNA ladder was detected in the rAAV-antisense 6A8 cDNA-transduced SKW6. Results from Annexin-V-FLUOS staining on anti-Fas antibody-treated cells revealed that the staining-positive rate in the rAAV-antisense 6A8 cDNA-transduced SKW6 cells was decreased in comparison to that in the wild-type and the mock-transduced cells. Giemsa’s staining observation showed that the number of dying (with apoptotic bodies) and dead cells was reduced in the rAAV-antisense 6A8 cDNA-transduced SKW6 cells in comparison with that in the wild-type and the mock-transduced cells upon anti-Fas antibody induction. The transduction did not affect the expression of Fas molecular on cell surface. 100% cells in all the groups showed Fas expression. The SKW6 cells became resistant to apoptosis induction by anti-Fas antibody upon transduction with a reverse fragment to a cDNA encoding human 6A8 α-mannosidase. The transduction did not affect the expression of Fas molecule on cells.     

Resistance of SKW6 cell to apoptosis induction with anti-Fas antibody upon transduction of a reverse fragment to a cDNA encoding human 6A8 α-mannosidase

The effect of transduction with a reverse fragment to a cDNA encoding human 6A8 ?-mannosidase on apoptosis induction of human B cell line SKW6 by anti-Fas antibody was tested. Apoptosis-inducer of anti-Fas monoclonal antibody was used to induce apoptosis in SKW6 cells. Giemsa’s staining, Annexin-V-FLUOS staining and DNA ladder test were used to determine the events of apoptosis. Indirect immunofluorescent staining with anti-Fas antibody was performed to detect the surface Fas expression. In a time-course test of 12, 24 and 36 h for apoptosis induction by anti-Fas antibody, DNA ladder was observed in the wild-type SKW6 cells in a time-dependent fashion. Mock transduction had no effect on DNA ladder production. However, no DNA ladder was detected in the rAAV-antisense 6A8 cDNA-transduced SKW6. Results from Annexin-V-FLUOS staining on anti-Fas antibody-treated cells revealed that the staining-positive rate in the rAAV-antisense 6A8 cDNA-transduced SKW6 cells was decreased in comparison to that in the wild-type and the mock-transduced cells. Giemsa’s staining observation showed that the number of dying (with apoptotic bodies) and dead cells was reduced in the rAAV-antisense 6A8 cDNA-transduced SKW6 cells in comparison with that in the wild-type and the mock-transduced cells upon anti-Fas antibody induction. The transduction did not affect the expression of Fas molecular on cell surface. 100% cells in all the groups showed Fas expression. The SKW6 cells became resistant to apoptosis induction by anti-Fas antibody upon transduction with a reverse fragment to a cDNA encoding human 6A8 a-mannosidase. The transduction did not affect the expression of Fas molecule on cells.     

S100B and S100A6 differentially modulate cell survival by interacting with distinct RAGE (receptor for advanced glycation end products) immunoglobulin domains

S100 proteins are EF-hand calcium-binding proteins with various intracellular functions including cell proliferation, differentiation, migration, and apoptosis. Some S100 proteins are also secreted and exert extracellular paracrine and autocrine functions. Experimental results suggest that the receptor for advanced glycation end products (RAGE) plays important roles in mediating S100 protein-induced cellular signaling. Here we compared the interaction of two S100 proteins, S100B and S100A6, with RAGE by in vitro assay and in culture of human SH-SY5Y neuroblastoma cells. Our in vitro binding data showed that S100B and S100A6, although structurally very similar, interact with different RAGE extracellular domains. Our cell assay data demonstrated that S100B and S100A6 differentially modulate cell survival. At micromolar concentration, S100B increased cellular proliferation, whereas at the same concentration, S100A6 triggered apoptosis. Although both S100 proteins induced the formation of reactive oxygen species, S100B recruited phosphatidylinositol 3-kinase/AKT and NF-kappaB, whereas S100A6 activated JNK. More importantly, we showed that S100B and S100A6 modulate cell survival in a RAGE-dependent manner; S100B specifically interacted with the RAGE V and C(1) domains and S100A6 specifically interacted with the C(1) and C(2) RAGE domains. Altogether these results highlight the complexity of S100/RAGE cellular signaling.     

Differentiated S100A7 expression in infected tonsils and tonsils from allergic individuals

Palatine tonsils are continuously exposed to microorganisms and antigens and secrete antimicrobial peptides as a first line of defense. S100A7 is a protein with antimicrobial and chemotactic properties. Our aim was to investigate how the expression of S100A7 in human palatine tonsils is affected by inflammatory processes. Tonsils obtained from 109 patients undergoing tonsillectomy were divided into groups of infected and noninfected as well as allergic and nonallergic, based on the results from tonsillar core culture tests and Phadiatop analysis, respectively. Western blot and immunohistochemistry were used to assess protein expression and real-time PCR was used to quantify mRNA levels. To explore the induction of S100A7, tonsils were stimulated with lipopolysaccharide in vitro. The immunohistochemical staining for S100A7 was most intense in the tonsillar epithelium, but the protein was also detected in B- and T-cell regions, which was confirmed with Western blot on isolated B and T cells. The S100A7 expression appeared to be the highest in CD8+ T cells. Reduced mRNA levels of S100A7 were detected in infected tonsils as well as in tonsils from allergic individuals. In vitro stimulation of tonsils with lipopolysaccharide did not have any effect on the expression. The results suggest a role for S100A7 in recurrent tonsillitis and allergic disease.     

S100A7 promotes the development of human endometriosis by activating NF-κB signaling pathway in endometrial stromal cells

Endometriosis (EM) is a chronic inflammatory disease affecting women aged between 23 and 42 years with a prevalence of 6%–10%. S100A7, a member of the S100 protein family, has been implicated in promoting inflammation. However, the role of S100A7 in EM and its underlying mechanism remain to be elucidated. S100A7 was silenced or overexpressed in primary endometrial stromal cells (ESCs). Cell proliferation was determined using a Cell Counting Kit-8. Cell cycle/apoptosis was monitored using a flow cytometer. Cell invasion was studied by a Transwell assay. Quantitative RT-PCR and Western blot analyses were used to evaluate gene expression. S100A7 and NF-κB expression is increased in both endometriotic tissue and ESCs from women with EM. The expression of S100A7 is correlated with the expression of NF-κB. S100A7 knockdown inhibits ESCs proliferation, cell cycle progression, cell invasion, and inflammation, but promotes cell apoptosis in an NF-κB dependent manner. In contrast, S100A7 overexpression demonstrated an inverse effect. S100A7 is increased in both endometriotic tissue and ESCs from women with EM. S100A7 overexpression contributes to EM through increasing ESCs proliferation, cell cycle progression, cell invasion, and inflammation, and inhibiting cell apoptosis in the NF-κB dependent manner. These findings highlight the importance of S100A7/NF-κB signaling in EM and provide new insights into therapeutic strategies for EM.