头颈部鳞癌生物标志物的预后作用

头颈部鳞癌(HNSCC)是世界上常见的恶性肿瘤之一。尽管现有治疗手段不断完善,HNSCC的5年生存率仍徘徊在50%。肿瘤生物标志物是由肿瘤细胞合成、释放或是宿主对肿瘤的反应而释放的物质。它有助于肿瘤的分子诊断、预后判断及疗效预测。因此生物标志物具有诊断、预后和预测三大作用。本文分别就与HNSCC相关的临床确定的和正在热研的,主要具有预后作用的生物标志物进行阐述。临床确定的标志物包括EBV和HPV。迄今为止众多生物标志物被研究且结果喜人。本文介绍的热研的标志物包括细胞周期调控、转录因子和信号传导三个方面。除了综述生物标志物预后作用的最新研究信息,还对研究中出现预后作用不一致的原因进行分析。分子细胞生物学的发展促使更多的生物标志物及潜在治疗靶点被发现。从预后到预测的发展是实施肿瘤个体化治疗的目标。以生物标志物为治疗靶点,有望改善HNSCC的预后,实现个体化精准治疗。     

单细胞测序技术在口腔鳞状细胞癌研究中的应用

单细胞测序技术是在单细胞水平上对目标细胞遗传信息进行测序。该技术已应用于众多医学领域,在研究肿瘤细胞的异质性、肿瘤的发生、发展及耐药性等方面具有独特的优势。口腔鳞状细胞癌是高侵袭性头颈部癌之一,其局部复发风险高,存活率低于一般头颈癌组,以手术治疗为主,放射和化学治疗为辅。然而,多数口腔鳞状细胞癌患者预后不佳,存在局部复发和高转移。本文总结了单细胞测序技术的发展及其在口腔鳞状细胞癌研究中的应用,旨在为口腔鳞状细胞癌的精准诊治提供新的见解。     

血常规在头颈部鳞状细胞癌早期诊断中的作用

目的:探究血常规指标在头颈部鳞状细胞癌早期诊断中的作用,从血液学的角度为头颈部鳞状细胞癌的早期诊断提供参考依据。方法:对181例头颈部良恶性肿瘤患者的血常规结果进行回顾性分析,比较头颈部鳞状细胞癌患者组(实验组)和头颈部良性肿瘤组(对照组)患者的血小板计数(PLT)、血小板比积(PCT)、血小板平均容积(MPV)、血小板分布宽度(PDW)、大血小板比率(P-LCR)、中性粒细胞计数(NEU)、淋巴细胞计数(LYM)、单核细胞计数(MONO)、嗜酸性粒细胞计数(EOS)、嗜碱性粒细胞计数(BASO)、血小板计数/淋巴细胞计数比值(PLR)以及中性粒细胞计数/淋巴细胞计数比值(NLR)共12个指标。结果:实验组和对照组的PLT、PCT、MPV、PDW、P-LCR以及BASO计数值比较均没有统计学差异(p0.05)。对照组NEU、MONO、EOS、PLR和NLR均低于实验组,而LYM计数值高于实验组,其差异均具有统计学意义(p0.05),ROC曲线显示NLR对于早期诊断的意义要优于PLR。结论:血小板相关指标在头颈部鳞状细胞癌早期诊断中无明显意义,白细胞亚型计数具有重要的提示作用,而NLR的诊断意义要高于PLR,或许可以联合其他鳞癌标志物进行早期诊断。     

CD9在口腔鳞状细胞癌中的表达水平及其临床意义

目的:口腔鳞状细胞癌是一类极易发生局部侵袭和淋巴结转移的恶性肿瘤,CD9蛋白在多种肿瘤的发生发展及侵袭转移过程中起到重要作用,本研究旨在分析CD9蛋白在口腔鳞状细胞癌中的表达水平及其临床意义。方法:收集我院诊断明确的口腔鳞癌肿瘤患者石蜡标本合计80例,通过免疫组化手段对CD9蛋白表达水平进行评价,并根据CD9蛋白的表达水平分组,分析患者的临床病理学特征与CD9蛋白的关系。结果:CD9在正常组织和癌旁组织正常表达,在肿瘤组织中表达率低,其表达水平和口腔鳞癌的分化程度,淋巴结转移及最终分期有相关性(P0.05)。结论:本研究结果揭示,CD9在口腔鳞状细胞癌的发生发展中起到重要作用,CD9蛋白水平的低表达或不表达可能预测着肿瘤具有更明显的恶性生物学行为,并可能成为口腔鳞状细胞癌预后的生物学指标及基因治疗的新靶点。     

CXXC4、EZH2在口腔鳞状细胞癌中表达及临床意义

目的:探讨口腔鳞状细胞癌(Oral Squamous Cell Carcinoma,OSCC)中CXXC指蛋白4(CXXC finger protein 4,CXXC4)与Zeste基因增强子同源物2(Enhancer of zeste homolog 2,EZH2)的表达、相关性及与临床病理特征之间的关系,为OSCC的早期诊断、治疗、判断预后及预防等提供参考依据。方法:采用免疫组织化学S-P法检测CXXC4蛋白及EZH2蛋白在60例口腔鳞状细胞癌组织及20例非肿瘤患者正常口腔黏膜组织中的表达情况,分析二者与口腔鳞状细胞癌临床病理特征的相关性。结果:口腔鳞状细胞癌组织中CXXC4蛋白的表达率为35.0%(21/60),明显低于正常口腔黏膜组织的60.0%(12/20)(P0.05);口腔鳞状细胞癌中EZH2蛋白的表达率为70.0%(42/60),高于正常口腔黏膜组织的25%(5/20)(P0.05)。CXXC4蛋白与EZH2蛋白的表达与口腔鳞状细胞癌的病理分化程度、TNM分期及淋巴结转移等密切相关(P0.05),而与年龄、性别及肿瘤直径大小等无关。CXXC4蛋白与EZH2蛋白在OSCC中表达呈负相关(r=-0.511,P0.001)。结论:CXXC4蛋白的表达下调或缺失以及EZH2蛋白的表达上调或过表达与口腔鳞状细胞癌的发生、发展及侵袭转移有关,可能作为口腔鳞状细胞癌预后预测的参考指标。     

鼻咽癌转移相关分子标志物的研究进展

鼻咽癌(Nasopharyngeal carcinoma NPC)是一种来源于鼻粘膜上皮细胞的恶性肿瘤,也是我国常见的头颈部恶性肿瘤之一.由于它的解剖位置相对隐匿,且多以低分化癌为主,因此大多数的鼻咽癌患者都死于肿瘤的转移而并非原发灶.研究表明,肿瘤的侵袭与转移与细胞的异常运动有关,是肿瘤细胞的粘附、降解、运动以及血管生成等多种生物学行为互相作用的动态过程.然而,影响NPC浸润转移的分子机制至今尚未完全阐明.随着现代分子生物学的发展,许多分子标志物被发现,使我们对鼻咽癌有了更高层次的认识.深入了解这些分子标志物的表达及调控,对筛选鼻咽癌标记物、提供药物治疗新靶点及预测治疗预后具有十分重要的意义.本研究拟对近年来鼻咽癌转移相关分子标志物的研究进展做一综述.     

microRNA在口腔鳞癌中的研究进展

口腔鳞状细胞癌(OSCC)是口腔颌面部恶性肿瘤中最主要的一类,约占80%以上,好发于男性,但近年来女性的发病率也呈现逐年增加的趋势;microRNA,是一类稳定的短序列非编码RNA,其主要功能是在转录后水平参与靶基因的调控,近来研究已发现在OSCC患者中存在许多异常表达的microRNAs,而这些分子在OSAS的发生发展中扮演着重要的角色,异常的microRNA同样对OSCC的临床诊断、治疗以及判断预后都有着重要的作用;本文对当前microRNA在OSCC中的的异常表达、作用机制以及作为诊断标记物、治疗靶点的潜能进行了综述。     

敲低脑内皮细胞粘附分子抑制HNSC细胞的侵袭能力

转移是包含头颈部鳞状细胞癌（head-and-neck squamous cell carcinoma, HNSC）在内的多种肿瘤复发和患者死亡的主要原因。目前,关于HNSC转移的网络调控机制仍有待进一步完善,以期降低HNSC死亡率。首先,通过在线数据库GEPIA统计后发现,脑内皮细胞粘附分子（cerebral endothelial cell adhesion molecule, CERCAM）在519例头颈部肿瘤中的相对表达量为4.98,是其在44例正常组织中（相对表达量为2.47）表达量的2.02倍。并且发现CERCAM表达量与头颈部肿瘤患者较差的预后正相关（P=0.015）。其次,在舌癌细胞SCC-9、喉癌细胞HEP2和鼻咽癌细胞CNE2敲低CERCAM的表达,发现上述3种细胞的侵袭能力均较对照组分别下降93.60%、37.18%和40.63%。最后,生物信息学预测结合Western印迹的结果证实,敲低CERCAM后,上调E-钙黏蛋白（E-cadherin）,同时下调锌指E盒结合蛋白(zinc-finger E-box-binding homeobox1, ZEB1)、波形蛋白(vimentin)和Twist相关蛋白1（Twist-related protein 1, TWIST1）。结果证实,CERCAM可能通过调控头颈部肿瘤细胞的上皮间充质转化（epithelial-mesenchymal transition, EMT）标志物来促进该类细胞发生侵袭。     

影响口腔鳞状细胞癌患者预后的临床病理因素分析

目的：研究影响口腔鳞状细胞癌（Oral squamous cellcar cinoma,OSCC）术后患者预后的临床病理因素。方法：回顾性研究55例手术治疗的原发口腔鳞状细胞癌患者与预后相关的因素：年龄、性别、发病部位、颈部淋巴结转移情况、肿瘤细胞分化程度等。结果：平均发病年龄为57.35±12.02岁,牙龈鳞癌的复发转移率最高（45.5%）,舌部第二（44%）,颊部第三（37.5%）,唇癌预后最好（0.0%）。术前颈部淋巴结转移情况、肿瘤细胞分化程度与预后有关。肿瘤细胞的分化程度与术前淋巴结转移无显著相关性。术前有颈部淋巴结转移合并中低分化与预后差相关。结论：口腔鳞状细胞癌的预后与发病部位无显著相关性。肿瘤中低分化及术前有淋巴结转移者易出现术后复发转移。     

影响口腔鳞状细胞癌患者预后的临床病理因素分析

目的:研究影响口腔鳞状细胞癌(Oral squamous cellcar cinoma,OSCC)术后患者预后的临床病理因素。方法:回顾性研究55例手术治疗的原发口腔鳞状细胞癌患者与预后相关的因素:年龄、性别、发病部位、颈部淋巴结转移情况、肿瘤细胞分化程度等。结果:平均发病年龄为57.35±12.02岁,牙龈鳞癌的复发转移率最高(45.5%),舌部第二(44%),颊部第三(37.5%),唇癌预后最好(0.0%)。术前颈部淋巴结转移情况、肿瘤细胞分化程度与预后有关。肿瘤细胞的分化程度与术前淋巴结转移无显著相关性。术前有颈部淋巴结转移合并中低分化与预后差相关。结论:口腔鳞状细胞癌的预后与发病部位无显著相关性。肿瘤中低分化及术前有淋巴结转移者易出现术后复发转移。     

mTOR信号通路及其在头颈肿瘤研究中的进展

哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,m TOR)在肿瘤的发生、侵袭、转移,甚至肿瘤的局部复发都起着重要的作用。头颈肿瘤作为全球最常见的癌症死亡原因之一,有多元的逐步恶化的过程。近年来研究表明m TOR信号通路的异常表达是头颈肿瘤最常见的病理改变之一,且m TOR信号通路的多种组成元件与头颈肿瘤患者的预后有明显的相关性,这不仅可为头颈肿瘤的特异性靶向治疗提供依据,而且也可为预测头颈肿瘤患者的预后提供参考。本文就m TOR信号通路及其在头颈肿瘤中的作用做一综述,旨在对m TOR信号通路和头颈肿瘤及其预后的关系有更进一步的认识。     

Plectin promotes migration and invasion of cancer cells and is a novel prognostic marker for head and neck squamous cell carcinoma

Head and neck squamous cell carcinoma (HNSCC) is usually found at a late stage and distant metastasis occurs at high frequency; therefore, novel prognostic markers are needed. This study was aimed to identify novel tumor markers in HNSCC. We identified 65 proteins which were significantly increased or decreased in the tumors by 2D-DIGE using 12 HNSCC and adjacent non-cancer tissues. Western blotting and immunohistochemical analysis confirmed that the expression of plectin was significantly increased in most cancer tissues as compared with non-cancer tissues. Strikingly, the suppression of endogenous plectin using siRNA inhibited the proliferation, migration and invasion of HNSCC cells and down-regulated Erk 1/2 kinase. Furthermore, immunohistochemistry using paraffin-embedded tissues from 62 patients showed not only that the frequency of recurrence was correlated with the plectin expression but that the prognosis of patients with a high plectin was extremely poor. Moreover, the survival rate of patients with a high plectin was significantly lower than that of patients with low E-cadherin levels, which is known to correlate with the poor prognosis of HNSCC. Our findings suggest that plectin promotes the migration and invasion of HNSCC cells through activation of Erk 1/2 kinase and is a potential prognostic biomarker of HNSCC.     

lncRNA在肿瘤中的表达及作用机制

长链非编码RNA(long non-coding RNAs,lncRNAs)是一类转录本长度大于200 nt,不具有蛋白编码功能的非编码RNA.近年来,随着高通量测序技术的发展,越来越多的功能性lncRNAs逐渐被发现,且已成为研究的热点.研究发现,lncRNAs可以作为致癌或抑癌基因参与肿瘤的发生发展.通过比对肿瘤细胞和正常细胞的表达谱显示,多种lncRNAs在不同类型肿瘤中异常表达.除了表型上的研究,目前已有部分报道深入研究了lncRNAs在肿瘤中的作用机制.例如,lncRNAs与肿瘤细胞凋亡、转移及耐药等过程密切相关.此外,在肿瘤患者的临床常规检验中检测到lncRNAs,提示其能够作为一种潜在的肿瘤诊断和预后因子. lncRNAs有望成为新型肿瘤标志物和肿瘤治疗的靶点,用于诊断、治疗、预测预后.本文将通过对lncRNAs在肿瘤中的作用及其分子机制进行综述.     

Tumor microenvironment – Unknown niche with powerful therapeutic potential

Head and neck squamous cell carcinomas (HNSCC) are in a group of cancers that are the most resistant to treatment. The survival rate of HNSCC patients has been still very low since last 20 years. The existence of relationship between oncogenic and surrounding cells is probably the reason for a poor response to treatment. Fibroblasts are an important element of tumor stroma which increases tumor cells ability to proliferate. Another highly resistance, tumorigenic and metastatic cell population in tumor microenvironment are cancer initiating cells (CICs). The population of cancer initiating cells can be found regardless of differentiation status of cancer and they seem to be crucial for HNSCC development.In this review, we describe the current state of knowledge about HNSCC biological and physiological tumor microenvironment.     

Overexpression of prothymosin alpha predicts poor disease outcome in head and neck cancer

Background

In our recent study, tissue proteomic analysis of oral pre-malignant lesions (OPLs) and normal oral mucosa led to the identification of a panel of biomarkers, including prothymosin alpha (PTMA), to distinguish OPLs from histologically normal oral tissues. This study aimed to determine the clinical significance of PTMA overexpression in oral squamous cell hyperplasia, dysplasia and head and neck squamous cell carcinoma (HNSCC).

Methodology

Immunohistochemistry of PTMA protein was performed in HNSCCs (n = 100), squamous cell hyperplasia (n = 116), dysplasia (n = 50) and histologically normal oral tissues (n = 100). Statistical analysis was carried out to determine the association of PTMA overexpression with clinicopathological parameters and disease prognosis over 7 years for HNSCC patients.

Results

Our immunohistochemical analysis demonstrated significant overexpression of nuclear PTMA in squamous cell hyperplasia (63.8%), dysplasia (50%) and HNSCC (61%) in comparison with oral normal mucosa (p_trend<0.001). Chi-square analysis showed significant association of nuclear PTMA with advanced tumor stages (III+IV). Kaplan Meier survival analysis indicated reduced disease free survival (DFS) in HNSCC patients (p<0.001; median survival 11 months). Notably, Cox-multivariate analysis revealed nuclear PTMA as an independent predictor of poor prognosis of HNSCC patients (p<0.001, Hazard''s ratio, HR = 5.2, 95% CI = 2.3–11.8) in comparison with the histological grade, T-stage, nodal status and tumor stage.

Conclusions

Nuclear PTMA may serve as prognostic marker in HNSCC to determine the subset of patients that are likely to show recurrence of the disease.     

Clinical and genomic characterization of neutral tumor evolution in Head and Neck Squamous Cell Carcinoma

Recent studies suggest that a significant proportion of cancers undergo neutral tumor evolution. We applied neutral evolution model in HNSCC patients from The Cancer Genome Atlas (TCGA). To ensure the accuracy of classification results, a sample with the purity of tumor <0.7 was excluded. A tumor sample was considered to evolve neutrally if R² ≥ 0.98. We found that about 16% of HNSCC patients undergo neutral tumor evolution. We showed that neutral evolution HNSCC patients have better prognosis and higher activities of immune response pathways, and the numbers of co-occurring mutation events and significantly positive selection mutations are significantly less than non-neutral evolution HNSCC patients. In conclusion, we described a comprehensive clinical and genomic characteristics of neutral tumor evolution in Head and Neck Squamous Cell Carcinoma (HNSCC), and provided evidence that the evolution history of HNSCC has both clinical and biological implications.     

Classification of DNA methylation patterns in tumor cell genomes using a CpG island microarray

Our group has initiated experiments to epigenetically profile CpG island hypermethylation in genomic DNA from tissue specimens of head and neck squamous cell carcinoma (HNSCC) using a microarray of 12,288 CpG island clones. Our technique, known as a methylation-specific restriction enzyme (MSRE) analysis, is a variation of the differential methylation hybridization (DMH) technique, in that it is not an array comparison of two DNA samples using methylation-specific restriction enzymes. Instead, it is a comparison of a single DNA sample's response to a methylation-sensitive restriction enzyme (HpaII) and its corresponding methylation-insensitive isoschizomer (MspI). Estimation of the reproducibility of this microarray assay by intraclass correlation (ICC) demonstrated that in four replicate experiments for three tumor specimens, the ICC observed for a given tumor specimen ranged from 0.68 to 0.85 without filtering of data. Repeated assays achieved 87% concordance or greater for all tumors after filtering of array data by fluorescence intensity. We utilized hierarchical clustering on a population of 37 HNSCC samples to cluster tumor samples with similar DNA methylation profiles. Supervised learning techniques are now being utilized to allow us to identify associations between specific epigenetic signatures and clinical parameters. Such techniques will allow us to identify select groups of CpG island loci that could be used as epigenetic markers for both diagnosis and prognosis in HNSCC.     

Microarrays and molecular markers for tumor classification

Human cancers have traditionally been classified according to their tissue of origin, histological characteristics and, to some extent, molecular markers. Clinical studies have associated different tumor classes with differences in prognosis and in response to therapy. Measurement of the expression of thousands of genes in hundreds of cancer specimens has begun to reveal novel molecularly defined subclasses of tumor; some of these classes appear to predict clinical behavior, while others may define tumor types that are ripe for directed development of therapeutics. Unfortunately, at present, differences between studies of similar tumor types can be as striking as their similarities.