cN0声门上型喉癌颈淋巴结隐匿性转移规律及处理

目的:探讨cN0声门上型喉癌患者颈部淋巴结隐匿性转移规律并选择合理的颈清扫区域。方法:139例cN0声门上型喉癌患者在行喉切除术同时行颈淋巴结清扫术,其中行改良性颈清扫57例,肩胛舌骨肌上淋巴结清扫30例,颈Ⅱ、Ⅲ区淋巴结清扫52例。将所获淋巴结按颈部分区逐一行组织病理学检查,观察其转移规律及临床疗效。结果:139例cN0声门上型喉癌患者中,同期行单侧颈清扫113例,同期行双侧颈清扫26例。139例(165侧)颈清扫标本经病理学检查,颈淋巴结阳性36例(25.9%),首次病理学检查颈淋巴结阴性者在随访中发现未手术侧淋巴结转移6例,总颈淋巴结隐匿性转移率为30.2%(42/139),单侧隐匿性转移率为26.6%(37/139),双侧隐匿性转移率为3.6%(5/139)。165侧颈清扫标本共获得淋巴结3 594枚,平均每侧21.8枚,共获病理阳性淋巴结83枚,其中位于Ⅰ区1枚(1.2%),Ⅱ区65枚(78.3%),Ⅲ区16枚(19.3%),Ⅳ区1枚(1.2%),Ⅴ区0枚。颈部复发率为5.0%(7/139),pN0与pN+的颈部复发率分别为0和16.7%(7/42),差异有统计学意义(P<0.05),总5年生存率为76.3%(106/139)。结论:颈Ⅱ、Ⅲ区是cN0声门上型喉癌颈部淋巴结隐匿性转移的主要区域,择区性(Ⅱ、Ⅲ区)颈淋巴结清扫术治疗cN0声门上型喉癌是合适的。     

CD105标记微血管密度与喉癌临床病理特征相关性

目的探讨喉鳞状细胞癌血管生成与各临床病理特征的关系。方法应用CD105单克隆抗体进行免疫组化SABC法检测54例喉癌组织中微血管密度(intratumormicrovesseldensity,IMVD)。结果54例喉癌组织中IMVD为8.854.50,T3和T4级喉癌组织IMVD(10.734.33)明显高于T1和T2级(6.803.68),其差异有显著性意义(P<0.01)。颈淋巴结转移组IMVD(12.873.39)明显高于非转移组(6.092.73),差异有显著性意义(P<0.01)。术后复发者组IMVD值(11.934.55)高于无复发者组(8.324.32),差别有意义(P<0.05)。结论CD105标记IMVD与喉癌T分级、颈淋巴结转移和肿瘤复发密切相关,可能是喉癌患者的一个独立的预后指标。     

声门上型喉癌颈淋巴结转移的临床病理研究

目的：观察声门上型喉癌颈淋巴结转移病理学特点，以及颈淋巴结转移和原发病变的关系。方法：１００例声门上型喉癌和颈廓清标本经火棉胶包埋、进行连续切片光镜观察。结果：①颈淋巴结转移５５例，转移率５５％；②颈淋巴结转移分四类：临床病理转移２９例，病理转移２６例，临床转移５例，无转移４０例；③转移淋巴结分四期：癌早期、癌长期、癌满期、破膜期；④转移淋巴结分三型：单发型、多发型和融合型；⑤声门上型喉癌不同发病率部位癌转移率为：杓会皱襞８５．７％，杓区６６．６％，会厌室带５６．８％，会厌４６．４％，室带４５．４％；⑥声门上型喉癌扩展到喉外者转移率为８０％～８４％。结论：声门上型喉癌有较高的颈淋巴结转移率，在转移淋巴结中融合型居多，破膜率高。分析肿瘤发病部位可帮助预测颈淋巴结转移。     

喉癌组织中水通道蛋白1的表达与微血管生成的相关性研究及意义

目的：明确水通道蛋白1（AQP1）在喉癌组织中的表达及分布与微血管形成的关系,并探讨其临床意义。方法：应用免疫组织化学技术检测喉癌组织及癌旁正常组织中AQP1蛋白的表达。通过CD105单克隆抗体标记肿瘤新生血管,检测肿瘤内微血管密度（IMVD）,并结合临床病理资料进行系统分析。结果：喉癌组织AQP1阳性细胞表达率和IMVD明显高于癌旁正常组织;AQP1的表达量随着喉癌分化程度的降低,其表达量逐渐升高;有淋巴结转移组AQP1的表达量明显高于无淋巴结转移组,差异有统计学意义。结论：①喉癌组织中AQP1的表达及IMVD与癌旁正常组织相比具有明显的特征性;②AQP1的高表达不仅与喉癌组织中IMVD呈明显的正相关性,且与喉癌有无淋巴结转移和细胞的分级密切相关;③推测AQP1在喉癌上皮和血管内皮细胞中表达增高能促进血管渗透性增强,在肿瘤的生长、浸润和转移中具有重要作用。     

声门上型喉癌临床N_0病理转移的观察

目的：为探讨喉癌颈淋巴结转移的规律，选择手术方法。方法：采用１１０例临床Ｎ０声门上型喉癌的１６４侧颈廓清术标本淋巴结连续切片方法观察。结果：发现颈淋巴结转移率为３５．５％（３９／１００）。提出临床Ｎ０病理转移的特点：１）转移淋巴结大小大多数介于０．５～１．５之间占８４．２％。２）转移淋巴结大多数为早期侵入期和生长发展期占８６．０％。３）转移淋巴结绝大多数为单发型占７４．４％。４）各个Ｔ分期均有转移淋巴结。结论：在颈淋巴结处理上，我们支持尽可能同期行选择性颈廓清术的观点     

声门上型喉癌颈淋巴结转移的临床病理研究

目的：观察声门上型喉癌颈淋巴结转移病理学特点，以及颈淋巴结转移和原发病变的关系。方法：１００例声门上型喉癌和颈廓清标本经火棉胶包埋、进行连续切片光镜观察。结果：①颈淋巴结转移５５例，转移率５５％；②颈淋巴结转移分四类：临床病理转移２９例，病理转移２６例，临床转移５例，无转移４０例；③转移淋巴结分四期：癌早期、癌长期、癌满期、破膜期；④转移淋巴结分三型：单发型、多发型和融合型；⑤声门上型喉癌不同发病     

p185和CD44v6与喉癌颈淋巴结转移的关系

目的 探讨癌基因蛋白P185和细胞黏附因子CD44v6与喉鳞状细胞癌（laryngeal squamous cell carcinOma，LSCC，简称喉癌），颈淋巴结转移的关系。方法 采用免疫组织化学SP法检测了63例喉癌中p185与CD44v6的表达。结果 p185在喉癌颈淋巴结转移组与非转移组间的表达存在差异（P〈0．05）：P185在不同病理分级（Ⅰ、Ⅱ、Ⅲ）组间存在差异（P〈0．05）：CD44v6在喉癌淋巴结转移组与非转移组间不同病理分级（Ⅰ、Ⅱ、Ⅲ）组间的表达无显著差异：喉癌中p185与CD44v6的表达存在正相关（r=-0．393，P=0．001）。结论 p185可能在喉癌的转移中起重要作用，与肿瘤细胞的恶性程度有关，CD44v6在喉癌转移中的作用不大，但两者在喉癌中可能存在协同作用。     

uPA基因启动子甲基化与喉鳞状细胞癌的关系

目的：探讨尿激酶型纤溶酶原激活剂（uPA）基因启动子甲基化状况与喉癌侵袭转移的关系。方法：采用RT-PCR法检测40例喉癌组织中uPAmRNA表达；同时用甲基化特异性PCR（MSP）法检测uPA基因启动子甲基化状况。结果：uPA基因表达率为65．0％（26／40）；有颈淋巴结转移的喉癌与无颈淋巴结转移的喉癌比较，uPA基因表达率显著增高（P〈0．01）。uPA基因启动子甲基化率为20．0％（8／40）；有颈淋巴结转移的喉癌与无颈淋巴结转移的喉癌比较，uPA基因启动子甲基化率显著降低（P〈0．01）。甲基化的喉癌组织中均无uPA基因表达。结论：uPA基因启动子甲基化是uPA基因表达缺失的机制之一，uPA基因启动子的去甲基化机制可能与喉癌颈淋巴结转移有关。     

CD44蛋白在人喉癌组织中的表达及临床意义

目的：探讨CD44在人喉癌组织中的表达及其临床意义。方法：采用免疫组织化学Envision法对261例喉不同病变组织标本进行CD44表达检测。结果：人喉癌组织中CD44高表达率为70．9％，明显高于癌前病变（16．7％）及正常喉组织（0％），其差异均有极显著性意义（P＜0．01）；喉癌组织中高分化鳞癌CD44表达明显低于低分化鳞癌（P＜0．01）；Ⅲ-Ⅳ期喉癌病变组织CD44表达明显高于I－Ⅱ期（P＜0．01）；有颈淋巴结转移的喉癌组织CD44表达明显高于无颈淋巴结转移的喉癌组织（P＜0．01）；CD44表达阳性病例3、5年生存率明显低于CD44表达阴性病例（均P＜0．05）。结论：喉癌的发生、发展及患者预后与CD44表达密切相关。     

颈部淋巴结处理方式对cN0声门上型喉癌患者预后的影响

目的:总结颈部淋巴结的处理方式对cN0声门上型喉癌患者预后的影响,探讨cN0声门上型喉癌患者行选择性颈部Ⅱ、Ⅲ和(或)Ⅳ区淋巴结清扫的意义。方法:回顾性分析我院2003-01-2007-05确诊为cN0声门上型喉鳞状细胞癌且有完整病历资料的83例患者,原发灶均经手术切除,随访5年以上或至患者死亡。分析择区性颈部淋巴结清扫术与其他方法(放疗、综合治疗、随诊观察)处理颈部淋巴结后,患者生存率有无统计学意义。结果:cN0声门上型喉癌颈部淋巴结转移率为30.77%,且随着T分期的增加,颈淋巴结的转移率也逐渐增高。采取干预措施的患者颈部淋巴结复发率明显低于未采取干预措施者(P<0.05)。清扫组与放疗组、综合组5年生存率差异均无统计学意义(P>0.05),观察组与清扫组差异有统计学意义(P<0.05)。结论:择区性颈清扫术是临床上处理cN0声门上型喉癌患者颈部淋巴结的有效措施之一。     

整合素αv亚基在喉和下咽鳞状细胞癌中表达及其与肿瘤血管发生的相关性

目的探讨整合素αv亚基在喉和下咽鳞状细胞癌（简称鳞癌）组织中的表达及其临床意义，以及整合素αv亚基表达与肿瘤血管发生的相关性。方法设计制作喉和下咽鳞状细胞癌组织芯片，应用免疫组化技术在组织芯片上检测整合素αv亚基和CD105的表达，根据CD105表达计算肿瘤新生血管密度，分析整合素αv亚基表达与喉和下咽鳞癌侵袭转移的关系，以及整合素αv亚基表达与肿瘤新生血管密度的相关性。结果喉和下咽原发鳞癌组织中整合素αv亚基阳性表达率为68．0％（51／75），明显高于癌旁正常组织（10．3％，3／29，Х^2=28．68，P〈0．001）；淋巴转移癌中整合素αv亚基表达率达100．0％（20／20），明显高于原发癌（Х^2=12．69，P〈0．05）和癌旁正常组织（Х^2=38．77，P〈0．001）；有淋巴转移组表达率明显高于无淋巴转移组（Х^2=10．87，P〈0．001）；整合素αv亚基的表达与肿瘤新生血管密度有明显相关性，阳性表达组的肿瘤微血管密度明显高于弱阳性组（q=3．31，P〈0．05）和阴性组（q=6．61，P〈0．001）；整合素αv亚基的表达率按照肿瘤的原发部位、分化程度、浸润范围以及患者的年龄、性别分组，差异均无统计学意义（P值均〉0．05）。结论整合素αv亚基的表达与喉和下咽鳞癌的淋巴转移关系密切，其过量表达可能通过诱发微血管形成而导致肿瘤发生侵袭和转移，整合素αv亚基有可能成为临床预测喉癌和下咽癌淋巴转移趋势的新型标记物。     

CD105与喉鳞状细胞癌血管生成及生物学行为的关系

目的：研究CD105在喉鳞状细胞癌血管中的表达，及其喉鳞状细胞癌生物学行为的意义。方法：采用双重免疫组织化学染色技术分别检测CD34／Ki67、CD105／Ki67在30例喉鳞状细胞癌和10例正常喉黏膜石蜡标本中的表达，分别以抗CD34和抗CD105单克隆抗体标记量化新生微血管（MVD），以抗Ki67单克隆抗体标记增殖的内皮细胞，并结合临床资料进行分析。结果：正常组和喉鳞状细胞癌组比较，MVDCD105差异有统计学意义（P〈0．05）；喉鳞状细胞癌组CD105标记的微血管增殖指数（PI）高于CD34标记组（P〈0．01）；MVDCD34与MVDCD105在喉鳞状细胞癌高分化组和低分化组的差异均有统计学意义（P〈0．05）；MVDCD34与MVDCD105均与淋巴结转移无关（P〉0．05）；MVDCD34与肿瘤的浸润范围无关（P〉0．05），与喉鳞状细胞癌的临床分期也无关（P〉0．05）；MVDCD105则随肿瘤浸润范围的增大而增加（P〈0．01），且随临床分期升高而增加（P〈0．01）。结论：CD105是新生血管的特异性标志物，新生血管密度与肿瘤的大小、临床分期相关，但与喉鳞状细胞癌的淋巴结转移无关。     

Immunohistochemical detection of cervical lymph node micrometastases from T2N0 tongue cancer

CONCLUSIONS: These results indicate that extensive, multiple cervical micrometastases occurred from an early stage in patients with T2N0 tongue cancer. The presence of micrometastases suggests the necessity of preventive neck dissection for Level I-IV nodes as a radical treatment. OBJECTIVE: Cervical lymph node metastases occur with a relatively high frequency in patients with T2N0 squamous cell carcinoma of the tongue, and control of the metastases greatly influences the prognosis of patients. In this study, micrometastases in the cervical lymph nodes were investigated to clarify the necessity and required extent of preventive neck dissection. MATERIAL AND METHODS: We investigated micrometastases in 24 subjects who had previously been diagnosed with T2N0 tongue cancer. We performed immunostaining with anti-cytokeratin antibody cocktail AE1/AE3 of sections of 401 paraffin-embedded lymph nodes obtained from these patients. RESULTS: Micrometastases were observed in 14 patients (58%) and were most abundant in Level II nodes (n=11; 46%). Micrometastases were observed in the Level IV nodes of 3 patients (13%), and upstaging to pN2b occurred in 7 patients (29%).     

喉癌组织中缺氧诱导因子1α和表皮生长因子受体的表达与微血管密度的关系

目的 分析喉癌组织中缺氧诱导因子1α(hypoxia-inducible factor-1α,HIF-1α)、表皮生长因子受体(epidermal growth factor receptor,EGFR)的表达与CD105标记计数微血管密度(micro vessel density,MVD)的关系及其在评价喉癌生物学行为和预后中的价值.方法 回顾性分析1990年1月至2002年1月91例喉癌患者临床病理资料及随访结果,免疫组化法检测肿瘤组织中HIF-1α、EGFR的表达,CD105标记肿瘤组织中微血管并计数MVD.结果 HIF-1α的表达与肿瘤大小、TNM分期、T分期、淋巴转移和病理分级有关(P值均<0.05);EGFR表达与TNM分期、淋巴转移、病理分级和复发有关(P值均<0.05);MVD与喉痛部位分型、TNM分期、T分期、淋巴转移、远处转移和病理分级有关(P值均<0.05).HIF-1α、EGFR表达水平与MVD有关(F值为7.644和5.197,P值为0.001和0.025).HIF-1α与EGFR表达的相关分析差异有统计学意义(r=0.238,P=0.007).患者3年和5年生存率为56.1%和44.2%,Log-rank法单因素生存分析表明喉癌部位分型、TNM分期、HIF-1α和EGFR的表达与患者预后有关(P值均<0.05);Cox比例风险模型分析显示TNM分期和EGFR的表达为影响喉癌患者预后的独立危险因素(P值分别为0.049和0.041,RR值分别为1.300和2.417).结论 在喉癌发生和进展过程中存在HIF-1α与EGFR的异常表达,且对调节肿瘤血管生成有一定的作用,并与肿瘤的生物学行为和预后有关.     

Determination of lymph node micrometastases in patients with supraglottic carcinoma

CONCLUSIONS: The frequency of loss of heterozygosity (LOH) at D9S 171 microsatellite locus on 9p21 may serve as an available method to evaluate occult micrometastases in laryngeal squamous cell carcinoma. High frequency of LOH was associated with a decreased probability of survival time. OBJECTIVE: To explore an available and sensitive method to detect cervical lymph node micrometastases in patients with laryngeal squamous cell carcinoma, the frequency of LOH at D9S171 microsatellite locus on 9p21 was studied. PATIENTS AND METHODS: Twenty samples from supraglottic cancer and 182 lymph nodes from neck dissections were examined by LOH comparing immunohistochemical (IHC) staining using cytokeratin 19 (CK19), and hematoxylin and eosin (H&E) staining. The frequency of lymph node metastasis and the clinical relevance were analysed. RESULTS: The frequency of LOH was 37.4% of lymph nodes and all of the primary tumors. Occult micrometastases were present in 9 of 20 cases; 23.6% of lymph nodes were positive for CK19 by IHC; 16.5% of lymph nodes were positive by H&E. There was a highly significant difference among the three methods. The highest rate of positive lymph nodes was at level II of the neck. There was a highly significant difference between overall survival time and lymph node metastasis with LOH and CK19 analysis.     

Detection of occult cervical micrometastases in patients with head and neck squamous cell cancer

OBJECTIVE: The incidence of occult nodal metastases associated with head and neck squamous cell carcinoma (HNSCC) and the clinical significance of nodal micrometastases by cytokeratin immunohistochemical analysis are examined. STUDY DESIGN: In all, 1012 lymph nodes from 50 patients treated between 1992 and 2001 at the University of Colorado Health Sciences Center (Denver, CO) were evaluated retrospectively for micrometastases. METHODS: Serial sectioning in 5-to 6-microm interval specimens stained either with hematoxylin and eosin (H&E) or immunostaining for cytokeratins using the monoclonal antibody cocktail AE1/AE3 was performed in 21 N0, 11 N1, and 14 N2 patient cases. Cases that showed scattered cells with suspect staining qualities but without morphological features consistent with HNSCC were further evaluated by epithelial membrane antigen (EMA) immunohistochemical analysis. RESULTS: H&E-stained and cytokeratin-stained sections revealed occult nodal micrometastases in 3.8% of N0 and 5% of N1 cases. Overall, 26 micrometastases were identified in N0 and N1 patients, causing 29% of N0 patients and 45% of N1 patients to be upstaged. Cytokeratin immunostaining detected micrometastases in eight cases that were negative on H&E serial sectioning. Serial sectioning by H&E alone identified three additional micrometastases. Negative EMA immunostaining confirmed the absence of malignant cells in lymph node sections that were equivocal on cytokeratin staining. CONCLUSIONS: The use of serial sectioning with H&E and cytokeratin immunohistochemical analysis increases the detection of micrometastases that are often elusive by routine processing in patients with HNSCC. Improved methods of detecting micrometastases may provide a basis for improved planning of postoperative therapy for patients already at risk for tumor recurrence.     

The Incidence of Micrometastases in Neck Dissection Specimens Obtained From Elective Neck Dissections

Although modern imaging techniques become more accurate for the assessment of lymph node metastases in the neck as criteria and technology evolve, micrometastases remain occult with any technique. Even the routine histopathological examination of neck dissection specimens is unable to detect all micrometastases. Because knowledge on the incidence of micrometastases in the clinically N0 neck might be of importance for decision making regarding elective treatment, a retrospective study on 96 elective neck dissections was conducted. Meticulous histopathological examination of the neck dissection specimens yielded 3092 lymph nodes of which 67 (2.2%) were tumor-positive. Twenty-six of these 67 lymph node metastases were micrometastases. Of the 36 tumor-positive neck dissection specimens, 21 contained micrometastases. In 9 tumor-positive specimens only micrometastases were found. This high incidence of micrometastases has important implications for the diagnostic work-up, the treatment, and histopathological examination of the N0 neck.     

Immunohistochemical studies in the identification of lymph node micrometastases in patients with squamous cell carcinoma of the head and neck

In the prediction of likely disease-free and overall survival intervals in patients with squamous carcinomas of the head and neck, cervical lymph node status assumes a prime role, and patients with cervical node metastases have diminished survivals, as a group, compared with patients whose cervical nodes are reported as negative for metastatic carcinoma. Conventional means of pathologic examination of cervical node biopsy specimens include examination of a single section through each individual node identified on gross examination, a process which, of necessity, leaves a significant portion of the node unexamined by microscopy. Recently, it has become apparent that more exhaustive pathologic sampling techniques, such as examining multiple sections of each lymph node, or staining each lymph node with antibodies to keratin via immunohistochemistry, will reliably yield a greater incidence of positive cervical lymph nodes ("micrometastases") than do conventional pathologic techniques. This suggests that the next line of inquiry should answer this question: just because micrometastases can be detected, should they be? Does the identification of (otherwise likely to be overlooked) tiny microscopic foci of spread of tumor in regional nodes by more sophisticated techniques yield additional data of real import to the patients, or is such information of lesser value? Should a role be defined in the care of head and neck cancer patients for the use of such advanced inquiries in the structuring of therapies, then the best approach to finding such elusive micrometastases (intraoperative immunohistochemistry? immunohistochemistry using routinely fixed tissues? polymerase chain reaction?) may subsequently be established.     

The significance of immunohistochemically demonstrated nodal micrometastases in patients with squamous cell carcinoma of the head and neck

OBJECTIVES/HYPOTHESIS: Patients with primary squamous cell carcinoma of the head and neck have a relatively high risk of occult lymph node metastases. Pathological demonstration of these metastases may be difficult, and the detection of such occult metastases may identify patients who are at an increased risk for early recurrence or reduced survival. Immunohistochemistry may be applied in the identification of occult metastases that may be missed on routine (H&E) histological examination. The aim of the study is to determine the prevalence and prognostic significance of immunohistochemically identified micrometastases in squamous cell carcinoma of the head and neck. STUDY DESIGN: A retrospective analysis of neck dissection specimens having no evidence of metastatic disease. METHODS: Lymph nodes from neck dissections performed on 10 patients with squamous cell carcinoma of the head and neck without conventional histological evidence of nodal metastases were subsequently stained for cytokeratins by the monoclonal antibody cocktail AE1/AE3 to detect micrometastases. RESULTS: Occult micrometastases were found in the lymph nodes 5 of 10 patients examined. There was no association between the site of primary tumor, or T tage, and the presence of occult metastases. Three of five patients found to have occult metastases developed recurrence in the neck, whereas only one of five patients with no evidence of micrometastases had regional recurrence. There was no significant discrepancy in the patient survival rate. CONCLUSIONS: Metastatic tumor cells are frequently present in lymph nodes, even in patients without histological evidence of nodal metastases by conventional methods. The presence of micrometastases may identify patients at increased risk for recurrence and may indicate poorer prognosis. The true clinical significance of these occult metastases will be determined by a long-term follow-up.