敲减TULP3对头颈鳞癌细胞生物学行为的影响

目的 探讨敲减TULP3对头颈鳞状细胞癌(HNSCC)细胞生物学行为的影响。方法 应用TCGA数据比较头颈鳞癌组织与癌旁组织中TULP3表达水平;体外培养人HNSCC细胞系HN4、HN6、CAL27、HSC3、SCC4及正常口腔上皮角质细胞HOK并应用Western Blot比较TULP3蛋白表达水平,免疫组化分析TULP3在HNSCC组织中表达情况。构建RNA干扰寡核苷酸si-TULP3及si-NC转染HN4、HN6,应用CCK-8实验、平板克隆形成实验、划痕愈合实验、Transwell侵袭实验检测敲减TULP3对HNSCC细胞增殖、侵袭、迁移能力的影响。实时定量逆转录PCR及Western Blot检测细胞周期及上皮间质转化(EMT)相关指标变化。构建HN6-shTULP3及HN6-shNC细胞株接种于裸鼠皮下,分析裸鼠移植瘤体积差异。结果 TCGA数据显示头颈鳞癌组织TULP3表达量显著高于癌旁组织(P<0.000 1);HN4、HN6、CAL27、HSC3细胞中TULP3蛋白表达量均高于HOK细胞,TULP3在HNSCC组织中呈阳性表达。HN4、HN6细胞转染si-TUL...     

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目的 探讨人头颈鳞癌中CCR7与磷酸化mTOR表达的关系,以及二者与肿瘤生物学行为之间的联系.方法 收集中国医科大学附属口腔医院口腔颌面外科2008-2009年收治的78例头颈鳞癌患者术后标本及临床详细资料,行免疫组化染色检测肿瘤原发灶CCR7和磷酸化mTOR表达情况,分析其表达的相关性及其与临床病理学因素间的关系.结果 78例头颈鳞癌原发灶中,CCR7阳性表达48例,磷酸化mTOR阳性表达40例;而10例正常口腔黏膜仅有1例CCR7阳性表达,无一例为磷酸化mTOR阳性表达.CCR7及磷酸化mTOR在头颈鳞癌原发灶与正常口腔黏膜中的表达差异均有统计学意义（均P＜ 0.05）.CCR7及磷酸化mTOR在不同临床分期和有无淋巴结转移患者的表达差异均有统计学意义（均P＜ 0.05）,而在不同年龄和性别患者的表达差异均无统计学意义(均P＞ 0.05).CCR7表达与磷酸化mTOR表达具有相关性(r=0.479,P＜0.05).结论 人头颈鳞癌组织中CCR7和磷酸化mTOR蛋白的高表达均与淋巴结转移和临床TNM分期相关,CCR7表达与磷酸化mTOR表达正相关.     

Stem cells in head and neck squamous cell carcinoma

The existence of a small subpopulation of tumourigenic cancer stem cells in the bulk of human head and neck squamous cancers (SCC) has been recognised in recent reports. This subpopulation has self-renewal properties and is responsible for the production of differentiated daughter cells that form the bulk of the tumour. Stem cells in head and neck SCC can be identified functionally using their self-renewal properties, or by their characteristic surface markers. As their resistance to contemporary cancer treatments may eventually lead to the failure of treatment there is an urgent need to better understand their biology with the ultimate goal of developing new diagnostic markers and curative cancer treatments.     

Apigenin inhibited hypoxia induced stem cell marker expression in a head and neck squamous cell carcinoma cell line

ObjectiveCancer stem cells contribute to tumor recurrence, and a hypoxic environment is critical for maintaining cancer stem cells. Apigenin is a natural product with anticancer activity. However, the effect of apigenin on cancer stem cells remains unclear. Our aim was to investigate the effect of apigenin on cancer stem cell marker expression in head and neck squamous cell carcinoma cells under hypoxia.DesignWe used three head and neck squamous cell carcinoma cell lines; HN-8, HN-30, and HSC-3. The mRNA expression of cancer stem cell markers was determined by semiquantitative RT-PCR and Real-time PCR. The cytotoxic effect of apigenin was determined by MTT colorimetric assay. Flow cytometry was used to reveal the number of cells expressing cancer stem cell surface markers.ResultsHN-30 cells, a cancer cell line from the pharynx, showed the greatest response to hypoxia by increasing their expression of CD44, CD105, NANOG, OCT-4, REX-1, and VEGF. Apigenin significantly decreased HN-30 cell viability in dose- and time-dependent manners. In addition, 40 μM apigenin significantly down-regulated the mRNA expression of CD44, NANOG, and CD105. Consistent with these results, the hypoxia-induced increase in CD44⁺ cells, CD105⁺ cells, and STRO-1⁺ cells was significantly abolished by apigenin.ConclusionApigenin suppresses cancer stem cell marker expression and the number of cells expressing cell surface markers under hypoxia.     

藤黄酸衍生物-5诱导人头颈部鳞状细胞癌细胞凋亡的体外研究

目的：：研究藤黄酸(GA)的衍生物5(C5),体外诱导人头颈部鳞状细胞癌(HNSCC)细胞的凋亡作用。方法：不同时间段分别用不同剂量C5作用于HNSCC细胞系CAL27、HN13和HN30,采用MTT法检测细胞存活率,Logit法测定抑制50%的细胞生长所需的浓度(IC50)值。采用Annexin-V/PI双染色流式细胞仪检测细胞凋亡。免疫细胞化学法和Western blot法检测凋亡相关蛋白表达的变化。结果：C5明显抑制CAL27、HN13和HN30细胞的生长,且与剂量和时间呈正相关。 IC50值比GA对照组明显降低。应用5.0μmol/L高浓度的C5,处理CAL27细胞系组24 h、48 h和72 h,细胞凋亡比例分别为52.19%、65.24%和84.53%。 C5引起明显的、剂量依赖性的Bcl-2表达减少和Bax表达增加。结论：C5可以体外通过上调Bax蛋白和下调Bcl-2蛋白表达诱导头颈部鳞状细胞癌细胞的凋亡。     

Toll样受体9与头颈部鳞状细胞癌关系的研究进展

Toll样受体(TLR)9与特异性配体结合后,不仅在包括头颈鳞状细胞癌(HNSCC)在内的多种恶性肿瘤的发生发展中高表达,而且具有促进肿瘤细胞增殖、侵袭及转移的作用;但TLR9在一些肿瘤中高表达预示较高的术后复发率及较低的术后生存率,在另一些肿瘤中高表达却是肿瘤预后较好的分子指标.TLR9参与HNSCC的相关分子机制,在于促进细胞增殖、抑制程序性细胞死亡、调节血管新生、促进肿瘤侵袭和促进免疫逃逸.目前,基于TLR9的抗肿瘤治疗已被广泛用于临床,其最终的临床效果尚待进一步了解TLR9的表达调控,分析TLR9参与HNSCC的双刃剑作用机制及相关信号转导环节,开发特异性高、不良反应小和可有效用于临床的TLR9佐剂或抑制剂.     

基于Oncomine数据库的数据挖掘在头颈鳞状细胞癌研究中的应用实例

目的：利用Oncomine数据库结合自身样本验证,获取靶分子钙调蛋白在头颈鳞状细胞癌（HNSCC）中的表达信息。方法???利用Oncomine数据库挖掘钙调蛋白在HNSCC中转录水平的变化,采用反转录聚合酶链反应、免疫印迹技术在本地区小样本队列中验证其转录及蛋白表达水平。结果钙调蛋白在Oncomine数据库大多数HNSCC队列中呈现转录水平的高表达,少部分呈现低表达;本地区样本结果显示,钙调蛋白在转录及蛋白表达水平均呈现高表达。结论??Oncomine数据库大样本量数据的挖掘并结合本地区样本的验证,能迅速准确地获取钙调蛋白在HNSCC中表达的相关信息,为后续的深入研究奠定基础。     

丙酮酸激酶M2与CD276在头颈鳞癌中的表达及临床意义

目的: 观察丙酮酸激酶M2（pyruvate kinase M2,PKM2）和B7H3（CD276）在头颈鳞癌（head and neck squamous cell carcinoma,HNSCC）和正常口腔黏膜中的表达,探讨PKM2与CD276的相关性。方法: 应用免疫组织化学方法检测PKM2和CD276在70例HNSCC及18例正常口腔黏膜组织中的表达水平,分析PKM2和CD276的表达水平与HNSCC临床病理特征的关系。采用SPSS 13.0软件包对数据进行统计学分析。结果: PKM2和CD276在HNSCC 的表达水平比正常黏膜组织高。PKM2和CD276的表达水平与TNM分期相关（P<0.05）,与患者性别、年龄、病理分级无相关性（P>0.05）。CD276在头颈鳞癌中的表达与PKM2的表达呈正相关。结论: PKM2和CD276在HNSCC的高表达与肿瘤恶性进展相关。PKM2可能通过增加CD276的表达而参与免疫调控。     

The role of SPP1 as a prognostic biomarker and therapeutic target in head and neck squamous cell carcinoma

Head and neck squamous cell carcinoma (HNSCC) is one of the most common malignancies and has a low 5-year survival rate. Mounting evidence suggests that oral potentially malignant disorders, such as oral leukoplakia (OLK), may progress to HNSCC. Given that OLK and HNSCC are often insidious and asymptomatic, the identification of markers of OLK malignant transformation and therapeutic targets in HNSCC is critical. Using various online tools and publicly available gene expression datasets, the secreted phosphoprotein 1 gene (SPP1) was identified as a significant differentially expressed gene among OLK, HNSCC, and non-cancerous tissues. SPP1 mRNA levels were elevated in HNSCC tissues and were associated with cancer stage, tumor grade, and human papillomavirus infection status. High SPP1 mRNA levels were correlated with poor overall survival of HNSCC patients. In contrast, SPP1 mutations were not significantly associated with overall survival, although their frequency in HNSCC was very low (0.6%). Furthermore, SPP1 expression levels in HNSCC were positively correlated with the infiltration of CD4⁺ cells, macrophages, neutrophils, and dendritic cells. The study results suggest that SPP1 may represent a diagnostic and prognostic biomarker, as well as a potential therapeutic target in HNSCC.     

Immunotherapy in the management of squamous cell carcinoma of the head and neck

Despite many advances in surgery, radiotherapy, and systemic treatments, only modest improvements in survival, function, and quality of life have been achieved after treatment of squamous cell carcinoma (SCC) of the head and neck. With a better understanding of the biology and genetics of tumours, the emergence of a paradigm shift towards the further development of non-surgical treatments may result in less morbidity and better outcomes than are seen currently. SCC of the head and neck is known to be a complex disease that has a sophisticated interaction with the human immune system. At the forefront of emerging treatments is immunotherapy, which has already been established in many other areas of oncology. The rapidly evolving nature of immunotherapeutic agents and, sometimes, their complex mechanisms can make the understanding of these concepts challenging, and could discourage clinicians from engaging in clinical trials. The aim of this paper therefore was to review the current premise for immunotherapeutic approaches, and to provide a contemporary evidence-based rationale for their use.