卵巢癌组织中Toll样受体9的表达及其临床意义

目的 检测Toll样受体9(TLR9)在卵巢癌组织中的表达,并探讨其临床意义.方法 采用免疫组织化学和Western blot方法检测30例卵巢癌组织及其癌旁组织、30例正常卵巢组织中TLR9的表达,并分析TLR9的表达与卵巢癌临床病理特征的关系.结果 免疫组织化学检测结果表明,卵巢癌组织、癌旁组织和正常卵巢组织中,TLR9蛋白的阳性表达率分别为80.0%、36.7%和20.0%,卵巢癌组织TLR9的阳性表达率高于正常卵巢组织和癌旁组织(均P＜0.01).TLR9蛋白的表达与肿瘤分化程度、国际妇产科协会(FIGO)分期、淋巴结转移有关(P＜0.05).Western blot检测结果显示,卵巢癌组织、癌旁组织和正常卵巢组织的相对表达量分别为0.803±0.072、0.411±0.087和0.113±0.065,卵巢癌组织中TLR9的表达量明显高于相应的癌旁组织及正常卵巢组织(均P＜0.01).结论 TLR9在卵巢癌组织中高表达,且TLR9的表达与卵巢癌的病理分级有关,提示TLR9可能通过免疫机制参与卵巢癌的发生、发展过程.     

Toll样受体-4与胰腺癌侵袭和转移的关系

背景与目的:Toll样受体-4(Toll-like receptor-4,TLR4)与多种肿瘤的侵袭和转移密切相关,然而其机制尚未阐明。本研究旨在探讨TLR4与胰腺癌侵袭和转移的关系,并分析其相关机制。方法:采用免疫组化法检测59例胰腺癌手术切除标本及30例癌旁正常胰腺组织中TLR4、基质金属蛋白酶-9蛋白(matrixmetalloproteinase-9,MMP-9)表达,分析TLR4、MMP-9蛋白表达与胰腺癌主要临床病理参数以及TLR4、MMP-9之间的相关性。以脂多糖(lipopolysaccharide,LPS)作用于体外培养的人胰腺癌细胞株PANC-1,通过Transwell小室检测细胞侵袭力,蛋白质印迹法(Western blot)检测MMP-9蛋白表达。结果:胰腺癌组织TLR4、MMP-9蛋白阳性表达率分别为72.9%和69.5%,均显著高于正常胰腺组织(P<0.01)。TLR4蛋白表达与肿瘤的TNM分期、淋巴结转移密切相关,MMP-9蛋白与肿瘤的分化程度、淋巴结转移及TNM分期密切相关(P<0.05)。Spearman等级相关分析表明,TLR4表达与MMP-9表达呈显著正相关(P<0.01)。经LPS作用后,PANC-1细胞的侵袭数目较正常组显著增加(P<0.01)。经LPS作用6、12和24 h后,PANC-1细胞MMP-9蛋白的相对表达量均显著高于正常组(P<0.01)。结论:TLR4蛋白表达与胰腺癌的侵袭和转移密切相关,其机制可能与促进MMP-9蛋白表达有关。     

Toll样受体2/4在非哺乳期乳腺炎组织中的表达及意义

目的探讨Toll样受体2(TLR2)和Toll样受体4(TLR4)在不同病理类型及不同临床分期非哺乳期乳腺炎(NPM)中的表达及其意义。 方法本研究为回顾性研究。选取2012年1月至2017年1月在南京医科大学附属淮安第一医院行手术治疗的141例NPM患者及10例乳腺纤维瘤患者(对照组)的病理切片进行TLR2、TLR4表达水平的免疫组织化学检测。根据术后病理结果将NPM分为肉芽肿性乳腺炎(GM)59例，浆细胞性乳腺炎(PCM)50例，其他类型乳腺炎32例，并将其中GM、PCM与对照组进行对比研究。根据临床分期将141例患者分为急性期(21例)、亚急性期(72例)、慢性期(48例)，并与对照组进行对比研究。通过检测TLR2、TLR4在不同病理类型及不同临床分期NPM中的表达水平，同时结合临床资料进行统计分析。多组间TLR2、TLR4表达水平的比较采用单因素方差分析，方差整齐时两两比较采用LSD法，方差不齐时两两比较采用Tamhane’s法；GM与PCM患者临床特征的比较采用χ²检验。 结果GM组TLR2、TLR4表达水平分别为15.82±4.96和27.27±7.70，PCM组TLR2、TLR4表达水平分别为15.29±4.14和26.25±6.63，对照组TLR2、TLR4表达水平分别为6.12±0.81和6.40±1.18。3组相比，TLR2、TLR4表达水平的差异均有统计学意义(F＝21.613、39.746，P均<0.001)，其中GM、PCM组TLR2、TLR4表达水平均高于对照组(P均<0.050)。并且，急性期、亚急性期、慢性期NPM及对照组患者相比，TLR2、TLR4表达水平的差异均有统计学意义(F＝190.112、246.965，P均<0.001)，其中急性期NPM患者TLR2、TLR4表达水平分别为23.65±2.32和40.10±2.22，高于亚急性期NPM的12.35±2.44和23.14±4.56(P均<0.001)，也高于慢性期NPM的17.19±2.36和29.36±2.17(P均<0.001)。与PCM患者相比，GM患者下肢结节红斑的发生率较高[28.8%(17/59)比12.0%(6/50)，χ²＝4.596，P＝0.032]。 结论TLR2和TLR4在急性期NPM中显著高表达，TLR2、TLR4信号途径可能参与NPM的发生、发展；GM与PCM患者的下肢结节红斑发生率存在差异。     

Toll样受体7及9在慢性粒细胞白血病患者浆细胞样树突状细胞内的表达

目的:研究Toll样受体7(Toll-like receptor 7,TLR7)mRNA及TLR9 mRNA在慢性粒细胞白血病(chronic myeloid leukemia,CML)患者外周血浆细胞样树突状细胞(plasmacytoid dendritic cell,pDC)内的表达及pDC分泌干扰素-α(interferon-α,IFN-α)的能力。方法:收集兰州大学第二医院血液科2010年11月至2011年7月间收治的30例CML患者(初诊未治组15例,缓解组15例)和体检中心的15例健康对照者,运用免疫磁珠法分选外周血pDC,利用real time-PCR检测pDC内TLR7及TLR9 mRNA表达水平。CpG ODN 2216刺激pDC 24 h后,ELISA检测上清液中IFN-α水平。结果:初诊组CML患者外周血pDC内TLR7 mRNA表达水平显著低于缓解组[(0.34±0.11)vs(0.93±0.21),P<0.05],初诊CML患者TLR9 mRNA表达水平显著低于缓解组[(0.44±0.15)vs(0.94±0.18),P<0.05]。CpG ODN 2216刺激后,初诊组pDC产生的IFN-α明显低于缓解组及健康对照组[(408.61±77.11)vs(611.39±84.86)、(651.67±93.39)ng/L,P<0.05]。结论:CML患者pDC内TLR7和TLR9 mRNA明显降低可能是pDC功能缺陷的主要原因,提示TLR7和TLR9可能参与CML的发病。     

肿瘤细胞固有Toll样受体9信号通路的研究进展

Toll样受体（TLR）9与其配体非甲基化胞嘧啶-鸟嘌呤二核苷酸序列的DNA（CpGDNA）特异结合后启动下游信号级联,TLR9信号活化受颗粒体蛋白、可溶性核酸感应蛋白（HMGBl）和可溶性TLR9调控。TLR9不再局限表达于免疫细胞,还在多系统肿瘤细胞中表达,并参与维系与促进肿瘤细胞的恶性生物学行为。肿瘤细胞固有TLR9信号通路功能的阐明将为肿瘤控制提供新思路。     

Toll样受体4与炎性肠病和结肠癌

Toll样受体(TLR)是天然免疫中重要的模式识别受体,主要在免疫细胞上表达,能特异性地识别病原相关分子模式,并在启动和调节天然免疫及获得性免疫中起到重要作用。研究表明TLR4在炎性肠病和结肠癌中表达增高。文章主要综述TLR4在结肠癌及炎性肠病癌变过程中的作用。     

Toll样受体和前列腺癌

Toll样受体（Tolllikereceptors,TLRs）是进化中比较保守的一个受体家族,至少包括10个成员;TLRs属模式识别受体（patternrecognitionreceptor,PRR）,不仅在激活天然免疫中发挥重要的作用,而且还调节获得性免疫,是连接天然免疫和获得性免疫的桥梁。近来发现,部分TLRs基因序列的多态性与前列腺癌的发生存在相关性;TLRs介导的信号通路能够活化树突状细胞,继而直接增强宿主对肿瘤天然免疫及获得性免疫应答的能力。我们围绕TLRs与前列腺癌的研究进展进行综述。     

Toll样受体与肿瘤预后相关性的Meta分析

目的:系统评价Toll样受体（Toll-like receptors,TLRs)在肿瘤组织中的表达水平及与肿瘤患者预后的相关性。方法:计算机检索中国期刊全文数据库（CNKI）、万方数据库（WF）、中文科技期刊数据库（VIP）、中国生物医学文献服务系统（Sinomed）、PubMed、The Cochrane Library、EMbase数据库,收集TLRs与肿瘤预后相关性的病例对照研究,检索时间为自建库起截止至2020年09月04日。根据纳入排除标准对文献进行筛选,经质量评价、数据提取后使用Stata 15.0软件对总生存期（overall survival,OS）、无进展生存期（disease-free survial,DFS）进行Meta分析。结果:最终纳入35篇文献,共4 472例患者,其中24篇涉及TLR4,7篇涉及TLR3,7篇涉及TLR9。结果表明,与对照组相比,TLR4阳性或高表达组肿瘤患者OS及 DFS较短（HR＝0.342,95％CI:0.098~0.586,P＝0.006;HR＝0.543,95％CI:0.278~0.809,P＝0.000）。TLR3阳性/高表达与晚期患者较好预后相关（HR＝-1.086,95％CI:-1.716~-0.455,P＝0.001）。TLR9阳性/高表达与早期患者的不良预后相关（HR＝0.656,95％CI:0.250~1.063,P＝0.002）。结论:不同亚型TLRs在肿瘤中发挥的作用不同,TLR4和TLR9阳性/高表达与肿瘤不良预后相关,TLR3阳性/高表达与晚期肿瘤患者较好预后相关,TLR3、TLR4、TLR9有望成为预测肿瘤预后的新型标志物。     

Toll 样受体4在恶性淋巴瘤中的作用

恶性淋巴瘤的发病机制一直是肿瘤研究的一个重要课题,但至今尚不完全明确。最近有临床研究表明TLR4（Toll－like receptor 4,TLR4）在套细胞淋巴瘤组织中和多种淋巴瘤细胞系中均有较高表达,且与预后有一定相关性。本文对TLR4在恶性淋巴瘤发生发展中的作用做一综述,以阐明淋巴瘤发病机制并为其治疗提供新的依据。     

Toll样受体基因多态性与EBV感染相关胃癌的易感性

目的： 探讨Toll样受体(TLR)家族成员TLR2基因启动子区-196~-174 del和TLR4基因 Thr399Ile位点多态性与EBV相关胃癌(EBVaGC)及EBV阴性胃癌(EBVnGC)易感性的关系。方法：采用PCR技术检测52例EBVaGC,157例EBVnGC以及94例正常对照人群TLR2(-196~-174 del)基因多态性;PCR-限制性片段长度多态性(PCR-RELP)技术检测50例EBVaGC,67例EBVnGC以及71例正常对照TLR4 Thr399Ile位点基因型及等位基因分布;分析2种基因多态性与EBVaGC及EBVnGC易感性的关系。结果：胃癌组与对照组比较TLR2(-196~-174 del)基因型分布无显著差异(χ2=3.180,P=0.075),胃癌组del等位基因频率明显高于对照组(χ2=4.875, P=0.027),del等位基因携带者的罹患危险性明显高于非携带者(OR=1.491,95%CI=1.045~2.126);EBVaGC组和EBVnGC组中,TLR2(-196~-174 del)3种基因型以及del等位基因频率差异无统计学意义(χ2=0.05,P=0.867)。EBVaGC组、EBVnGC组和正常对照组中均未发现TLR4基因Thr399Ile位点的多态,其基因型分布及等位基因频率差异均无统计学意义(P>0.05)。结论：TLR2(-196~-174 del)等位基因可能是胃癌发病危险因素,且在EBVaGC和EBVnGC两种胃癌发生中产生相同的影响。未发现TLR2(-196~-174 del)和TLR4基因Thr399Ile基因型分布与EBVaGC的易感性相关。     

High expression of Toll-like receptor 4/myeloid differentiation factor 88 signals correlates with poor prognosis in colorectal cancer

Background:

The Toll-like receptor (TLR) 4 signalling pathway has been shown to have oncogenic effects in vitro and in vivo. To demonstrate the role of TLR4 signalling in colon tumourigenesis, we examined the expression of TLR4 and myeloid differentiation factor 88 (MyD88) in colorectal cancer (CRC).

Methods:

The expression of TLR4 and MyD88 in 108 CRC samples, 15 adenomas, and 15 normal mucosae was evaluated by immunohistochemistry, and the correlations between their immunoscores and clinicopathological variables, including disease-free survival (DFS) and overall survival (OS), were analysed.

Results:

Compared with normal mucosae and adenomas, 20% cancers displayed high expression of TLR4, and 23% cancers showed high expression of MyD88. The high expression of TLR4 and MyD88 was significantly correlated with liver metastasis (P=0.0001, P=0.0054). In univariate analysis, the high expression of TLR4 was significantly associated with shorter OS (hazard ratio (HR): 2.17; 95% confidence interval (95% CI): 1.15–4.07; P=0.015). The high expression of MyD88 expression was significantly associated with poor DFS and OS (HR: 2.33; 95% CI: 1.31–4.13; P=0.0038 and HR: 3.03; 95% CI: 1.67–5.48; P=0.0002). The high combined expression of TLR4 and MyD88 was also significantly associated with poor DFS and OS (HR: 2.25; 95% CI: 1.27–3.99; P=0.0053 and HR: 2.97; 95% CI: 1.64–5.38; P=0.0003). Multivariate analysis showed that high expressions of TLR4 (OS: adjusted HR: 1.88; 95% CI: 0.99–3.55; P=0.0298) and MyD88 (DFS: adjusted HR: 1.93; 95% CI: 1.01–3.67; P=0.0441; OS: adjusted HR: 2.25; 95% CI: 1.17–4.33; P=0.0112) were independent prognostic factors of OS. Furthermore, high co-expression of TLR4/MyD88 was strongly associated with both poor DFS and OS.

Conclusion:

Our findings suggest that high expression of TLR4 and MyD88 is associated with liver metastasis and is an independent predictor of poor prognosis in patients with CRC.     

维吾尔族妇女宫颈癌病理进程与Toll样受体9蛋白表达的关系

目的:Toll样受体9(Toll-like receptor-9,TLR9)是天然免疫受体之一,近期研究表明TLR9与多种上皮性肿瘤发生的早期行为密切相关。本研究拟从蛋白质水平观察宫颈上皮组织中TLR9表达变化与维吾尔族妇女宫颈癌病理进程的关系。方法:收集维吾尔族妇女宫颈炎、宫颈上皮内瘤变cervical intraepithelial neoplasia,CIN)和宫颈鳞癌(cervical squamous cell carcinoma,CSCC)患者的宫颈上皮石蜡包埋组织标本共97例,采用免疫组织化学方法检测各组织中TLR9蛋白表达水平。结果:TLR9蛋白在绝大部分宫颈炎患者的宫颈上皮细胞中不表达(88.2%)或弱表达(11.8%),但是随着宫颈上皮内瘤变和宫颈癌的发生,表达水平逐渐上升,其阳性率依次为慢性宫颈炎(2/17,11.8%),CINⅠ(4/19,28.6%),CINⅡ(3/10,30.0%),CINⅢ(12/24,50.0%),CSCC(17/32,53.1%),5组之间比较差异有统计学意义(P0.05),CINⅢ及CSCC与宫颈炎间的阳性率差异显著(P0.05),但是CINⅠ和CINⅡ与宫颈炎间的差异无统计学意义(P0.05),并且在CSCC中,不同肿瘤分化程度之间其TLR9蛋白阳性率的差异无统计学意义(P0.05)。结论:TLR9蛋白可能参与了宫颈癌的发生和发展的进程。     

Regulation of migration and invasion by Toll-like receptor-9 signaling network in prostate cancer

Toll-like receptors (TLRs) play an important role in tumorigenesis and progress of prostate cancer. However, the function and mechanism of Toll-like receptor-9 (TLR9) in prostate cancer is not totally understood. Here, we found that high expression of TLR9 was associated with a higher probability of lymph node metastasis and poor prognosis. Further in vitro functional study verified that silence of TLR9 inhibited migration and invasion of PC-3 cells, indicating expression of TLR9 involving in the migration and invasion of cancer cells. The data of microarray exhibited silence of TLR9 induced 205 genes with larger than 2-fold changes in expression levels, including 164 genes down-regulated and 41 genes up-regulated. Functional Gene Ontology (GO) processes annotation demonstrated that the top three scores of molecular and cellular functions were regulation of programmed cell death, regulation of locomotion and response to calcium ion. TLR9 signaling network analysis of the migration and invasion related genes identified several genes, like matrix metallopeptidase 2 (MMP2), matrix metallopeptidase 9 (MMP9), chemokine receptor 4 (CXCR4) and interleukin 8 (IL8), formed the core interaction network based on their known biological relationships. A few genes, such as odontogenic ameloblast-associated protein (ODAM), claudin 2 (CLDN2), gap junction protein beta 1 (GJB1) and Rho-associated coiled-coil containing protein kinase 1 pseudogene 1 (ROCK1P1), so far have not been found to interact with the other genes. This study provided the foundation to discover the new molecular mechanism in signaling networks of invasion and metastasis in prostate cancer.     

肺癌组织中Toll样受体9的表达及其对术后放疗生存的影响

目的: 分析肺癌组织中Toll样受体9(TLR9)表达与肺癌临床病理特征的关系及其对肺癌术后放疗结果的影响。方法:收集2007年3月至2011年12月经本院手术治疗的肺癌病史资料63例,采用免疫组化SP法检测癌组织中TLR9的表达,分析TLR9表达与肺癌临床病理特征的关系;通过对36例肺癌术后放疗患者进行Kaplan-Meier单因素生存分析和多因素Cox回归分析,探讨TLR9表达在肺癌术后放疗生存中的意义。结果:正常肺泡细胞内未见TLR9蛋白阳性表达,而TLR9蛋白定位于肺癌组织的肿瘤细胞质中,阳性表达率为68.3%(43/63)。且随着肿瘤直径增加,TLR9表达阳性率增加(PPPP结论:肺癌组织中TLR9表达阳性率升高,TLR9表达和淋巴结转移是肺癌术后放疗患者生存的独立危险因素,TLR9可能成为预测肺癌术后放疗生存的分子生物学指标。     

TLR4 Thr399Ile基因多态性与胃癌易感性的研究

目的:探讨Toll样受体4（toll-like receptor 4,TLR4）Thr399Ile基因多态性与胃癌易感性的关系。方法:通过计算机检索及手工检索,收集有关TLR4 Thr399Ile基因多态性与胃癌易感性的文献,筛选出符合纳入和排除标准的文献,应用Meta分析软件对各项研究进行异质性检验,计算合并OR值及其95％CI,并行敏感性分析和发表偏倚的评估。结果:8篇文献纳入本研究,共计有1297例胃癌患者和2702例对照人群。TLR4 Thr399Ile基因多态性与胃癌易感性的研究结果为:T versus C:OR=1.307,95%CI=1.047-1.631;TT versus CC:OR=1.527,95%CI＝0.398-5.860;TC versus CC:OR=1.324,95%CI＝1.049-1.670;TT/TC versus CC:OR=1.326,95%CI＝1.053-1.671;TT versus TC/CC:OR=1.481,95%CI＝0.386-5.685。根据种族来源进行分层分析,在高加索人群中的研究结果为:T versus C:OR=1.298,95%CI＝1.027-1.640;TC versus CC:OR=1.313,95%CI＝1.027-1.679;TT/TC versus CC:OR=1.316,95%CI＝1.031-1.679。结论:TLR4 Thr399Ile TC、TT/TC基因型及T等位基因能增加胃癌的患病风险。     

Toll-like receptor 9 agonist IMO cooperates with everolimus in renal cell carcinoma by interfering with tumour growth and angiogenesis

Background:

Targeting the mammalian target of rapamycin by everolimus is a successful approach for renal cell carcinoma (RCC) therapy. The Toll-like receptor 9 agonist immune modulatory oligonucleotide (IMO) exhibits direct antitumour and antiangiogenic activity and cooperates with both epidermal growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF) inhibitors.

Methods:

We tested the combination of IMO and everolimus on models of human RCC with different Von-Hippel Lindau (VHL) gene status, both in vitro and in nude mice. We studied their direct antiangiogenic effects on human umbilical vein endothelial cells.

Results:

Both IMO and everolimus inhibited in vitro growth and survival of RCC cell lines, and their combination produced a synergistic inhibitory effect. Moreover, everolimus plus IMO interfered with EGFR-dependent signaling and reduced VEGF secretion in both VHL wild-type and mutant cells. In RCC tumour xenografts, IMO plus everolimus caused a potent and long-lasting cooperative antitumour activity, with reduction of tumour growth, prolongation of mice survival and inhibition of signal transduction. Furthermore, IMO and everolimus impaired the main endothelial cell functions.

Conclusion:

A combined treatment with everolimus and IMO is effective in VHL wild-type and mutant models of RCC by interfering with tumour growth and angiogenesis, thus representing a potentially effective, rationale-based combination to be translated in the clinical setting.     

Association study of a functional Toll-like receptor 4 polymorphism with susceptibility to gastric mucosa-associated lymphoid tissue lymphoma

Toll-like receptor 4, as part of innate immune response, is the main receptor for lipopolysaccharide on marginal zone B cells. The rare allele of TLR4 Asp299Gly attenuates receptor signaling and diminishes the inflammatory response. We genotyped 87 patients with gastric mucosa-associated lymphoid tissue (MALT) lymphoma, 594 Helicobacter pylori positive controls and 358 healthy blood donors to investigate an association of TLR4 Asp299Gly in the development of gastric MALT lymphoma. Heterozygote genotype was significantly less frequent in patients with gastric MALT lymphoma compared to H. pylori-infected controls (4.6% vs. 11.6%, Fischer's exact P = 0.019, odds ratio = 0.37, 95% confidence interval 0.13 - 1.03). Because 10% of caucasians are carriers of the rare allele G TLR4 Asp299Gly appears to be only one factor in the genetic susceptibility to gastric lymphoma. Further studies in larger samples are needed to confirm our findings and fully elucidate the role of TLR4 and its genetic variants in the pathophysiology of H. pylori infection and gastric lymphoma.     

Gastric extranodal marginal zone B-cell lymphomas of MALT type exclusively express Toll-like receptor 4 in contrast to other lymphomas infiltrating the stomach.

BACKGROUND: Development and growth of extranodal marginal zone B-cell lymphomas (eMZBCLs) of mucosa-associated lymphoid tissue (MALT) type are thought to be highly dependent on Helicobacter pylori and autoantigens. Receptors mediating these effects are not characterised so far. Toll-like receptors (TLRs) recognise bacterial proteins and autoantigens, which results in inflammatory reactions and influences tumour development and growth. MATERIALS AND METHODS: TLR4, 5 and 9 expressions were evaluated by immunohistology and confocal microscopy in gastric eMZBCL in comparison to other lymphomas infiltrating the stomach. RESULTS: TLR4 was exclusively expressed on the cell surface in all eMZBCL (n = 19) and not in chronic lymphocytic leukaemia (CLL, n = 12) or mantle cell lymphoma (MCL, n = 10). TLR5 was strongly expressed in CLL and weak in some eMZBCL (15 of 19), but not in MCL. TLR4, 5 and 9 were negative in all the three lymphoma entities. CONCLUSIONS: Exclusive TLR4 expression may enable eMZBCL to interact with H. pylori and autoantigens. Blockade of TLR4 might be a new approach for therapy of eMZBCL of MALT type.     

TLR4与肿瘤免疫逃逸的研究进展

Toll样受体(Toll-like receptor, TLR)家族是感受病原体入侵的一类模式识别受体,目前在哺乳动物免疫系统中共发现11个成员,其中以TLR4最受关注。大部分肿瘤组织均表达TLR4;肿瘤细胞上TLR4激活后能以不同方式促进肿瘤的发生、发展、凋亡抵抗和侵袭、转移;TLR4参与肿瘤免疫逃逸除了外源性活化机制外,肿瘤微环境中的内源性配体同样可能发挥重要作用。TLR4有望成为肿瘤生物治疗的新靶点。