喉鳞癌中P糖蛋白表达与细胞增殖、凋亡及预后的关系

目的 探讨P糖蛋白在喉鳞癌组织中表达与肿瘤细胞的增殖、凋亡活性,及其临床病理特征与患者预后的关系。方法 应用免疫组化技术检测86例患者喉鳞癌组织中P糖蛋白、Ki 67的表达,TUNEL法测定喉鳞癌细胞凋亡率。结果 P糖蛋白在喉鳞癌组织中的阳性表达率为39.5％(34/86),与喉鳞癌的临床分期、组织学分级、淋巴结转移及患者术后生存率有关(均P＜0.05);P糖蛋白表达阳性组的肿瘤细胞凋亡率比阴性组低(P＜0.05),而增殖率差异无统计学意义(P＞0.05)。结论 P糖蛋白可能对喉鳞癌细胞的凋亡活性具有一定抑制作用;P糖蛋白阳性表达率对于评估喉鳞癌病程进展及预后具有一定价值。     

EphA2调控头颈部鳞状细胞癌血管新生及转移的体内实验

目的 研究EphA2对头颈部鳞状细胞癌(简称鳞癌)血管新生及颈淋巴转移的影响.方法 利用慢病毒介导的短发夹RNA( short hairpin RNA,shRNA)沉默EphA2在高转移性头颈鳞癌细胞株M2中的表达,嘌呤霉素筛选稳定克隆,反转录PCR及Western blot技术检测EphA2沉默效果;构建头颈鳞癌高转移动物模型,通过HE染色验证其成瘤及颈淋巴转移情况;免疫组织化学技术检测移植瘤微血管密度,Western blot技术检测移植瘤标本中EphA2及血管内皮生长因子(VEGF)的表达情况.结果 筛选出EphA2基因稳定沉默的M2细胞(定义为M2 EphA2 RNA+即实验组),并成功构建头颈鳞癌高转移动物模型,成瘤率达100％.25 d后,实验组移植瘤体积为(430.7±190.0) mm3(x±s,以下同)较对照组的(1179.0 ±289.4) mm3明显减少(t =5.597,P＜0.01),质量显著减轻(t=-4.560,P＜0.05),且双侧颈淋巴转移率明显降低(Mann-Whitney U=10.0,P＜0.05).Western blot 显示实验组移植瘤组织中EphA2及VEGF蛋白表达显著下调,免疫组织化学技术发现实验组瘤体微血管密度为(4.74±0.67)个/视野,显著少于对照组的(14.17±0.59)个/视野(t=26.751,P＜0.01).结论 EphA2沉默能抑制头颈部鳞癌细胞的生长及转移,并能明显减少头颈部鳞癌组织内的肿瘤新生血管,且该肿瘤新生血管的抑制可能与促血管生成因子VEGF的减少有关.     

增殖细胞核抗原与喉癌生物学行为相关性的研究

目的探讨增殖细胞核抗原(PCNA)表达与喉癌生物学行为的相关性.方法采用流式细胞免疫法检测40例喉鳞状细胞癌患者PCNA的表达,同时检测8例癌旁组织作为对照.结果高中分化鳞癌与低分化鳞癌差异有高度显著性(P＜0.01),淋巴结转移阳性组与阴性组差异有显著性(P＜0.05),早期组(I,Ⅱ)与晚期组(Ⅲ,Ⅳ)差异无显著性(P＞0/05),癌组织与癌旁组织间PCNA指数差异有高度显著性(P＜0.001).结论PCNA表达与喉癌的病理分级、淋巴结转移呈显著正相关,与临床分期无关,PCNA高表达是喉癌的重要生物学特征.     

Jab1基因沉默对裸鼠人喉癌移植瘤生长影响的研究

目的 探讨Jab1重组质粒对人喉鳞状细胞癌(简称鳞癌)Hep-2细胞裸鼠移植瘤生长的影响.方法 利用喉鳞癌Hep-2细胞裸鼠皮下移植瘤模型,通过向瘤体内注射pJab1、阴性对照质粒和生理盐水,观察瘤体增长情况,通过免疫组化方法、反转录聚合酶链反应( RT-PCR)观察瘤体内Jab1、p27的mRNA及蛋白表达水平的变化.结果 pJab1质粒组的肿瘤体积为(267.60±88.19)mm3((x)±s,以下同),与阴性对照组的(832.20±140.39) mm3及生理盐水组的( 895.40±145.93) mm3相比,差异有统计学意义(F=36.73,P＜0.001);免疫组化结果显示pJab1质粒组瘤内的Jab1蛋白的表达率降低为32.40％±5.59％,p27蛋白的表达率增高到76.80％±6.30％(P＜0.001),与阴性对照组及生理盐水组比较,差异有统计学意义;RT-PCR结果显示实验组瘤内Jab1的mRNA相对表达水平降低至0.65±0.03(F=558.00,P＜0.001),p27的mRNA无明显变化,为0.80±0.02(F=1.52,P＞0.05).结论 pJab1干扰质粒可明显降低裸鼠肿瘤组织内Jab1基因的表达并抑制瘤体的生长;pJab1质粒能有效抑制Jab1基因的表达,有望成为临床治疗喉癌的新途径.     

细胞骨架分子与喉鳞状细胞癌患者复发及预后的关系

目的 观察细胞骨架分子Fascin-1,Ezrin及Paxillin在喉鳞状细胞癌(简称鳞癌)组织中的表达,并评价三者对喉鳞癌复发风险及预后的预测价值.方法 采用免疫组化Max Vision二步法观察三种蛋白在199例患者喉鳞癌组织中的表达情况,通过非条件Logistic回归及单因素、多因素生存分析,总结三者在喉鳞癌复发风险预测及预后评估中的价值.结果 三种蛋白分别在低分化、颈淋巴转移及临床晚期(Ⅲ+Ⅳ)喉鳞癌组织中表达增强趋势(P值均＜0.05),且三者表达呈正相关趋势(P值均＜0.001).三种蛋白分别在复发组中的表达强度高于未复发组(x2值分别为42.479,43.673和22.261,P＜0.05).三种蛋白联合高表达患者中复发病例构成比为69.1％,高于其他联合表达模式组中的构成比(P ＜0.001),且Fascin-1(比值比为7.89,95％可信区间为2.26～27.53,P=0.001)及Ezrin(比值比为2.51,95％可信区间为1.18 ～5.32,P＜0.001)为影响喉鳞癌复发的独立危险因素.三种蛋白分别高表达患者的5年无瘤生存率均降低(P＜0.05),而三者联合高表达患者5年无瘤生存率更低,为26.4％ (P＜0.05).三种蛋白均为影响患者预后独立危险因素(P＜0.05).结论 Fascin-1,Ezrin,Paxillin不仅与喉鳞癌的恶性进展相关,还与复发风险及预后不良相关.联合检测三者表达情况,有助于喉鳞癌复发风险预测及预后评估.     

喉鳞状细胞癌组织体外原代培养的初步研究

目的 应用体外培养技术,对喉鳞状细胞癌组织进行体外培养,探讨喉鳞癌组织原代培养中的各种影响因素,为建立人喉鳞癌组织的细胞系提供实验基础。方法采用体外组织培养技术,对24例人喉鳞状细胞癌组织进行原代培养,观察原代培养中肿瘤细胞的生长与供体的年龄、肿瘤组织的分化程度及不同培养方法的关系,分析在人喉鳞癌细胞的培养中成纤维细胞、微生物污染的影响。结果 24例人喉鳞状细胞癌组织标本,年龄小于60岁组的细胞生长率为31.25％(5/16例),年龄大于60岁组为37.5％(3/8例);高分化组为100％(2/2例),中分化组为30.8％(4/13例),低分化组25％(2/8例);组织块培养法为43.75％(7/16例),酶消化法培养为10％(1/10例);倒置显微镜下观察,在培养的第5～7天,在贴壁组织块周围可见到有上皮样细胞爬出。全部标本中,成纤维细胞的过度生长和微生物的污染是阻碍人喉鳞癌细胞生长的重要因素。结论培养组织的细胞生长率与供体的年龄关系不大;肿瘤组织的分化程度较高者,细胞的生长率较高;与酶消化分离培养法相比,贴壁组织块培养法的细胞生长率较高;成纤维细胞及微生物的污染是阻碍人喉鳞癌细胞系建立的重要因素。     

喉鳞状细胞癌组织中人白细胞I类分子和抗原加工递呈分子的表达

目的 研究喉鳞状细胞癌(简称鳞癌)组织中人白细胞抗原(human leucotyte antigen,HLA)Ⅰ类分子和抗原加工递呈(antigen processing machinery,APM)分子抗原转运相关蛋白1(transporter associated with antigen processing,TAP-1)和低分子量蛋白7(low molecular weight polypeptide,LMP-7)下调的频率及其临床意义.方法 用免疫组化方法检测2001-2002年手术切除的51例喉鳞癌组织中HLA-I类抗原重链HLA-ABC和APM分子TAP-1、LMP-7表达情况,并分析其与临床病理特征及预后的关系.结果 HLA-ABC、TAP-1、LMP-7各分子在喉鳞癌组织中的下调率分别是56.9％(29/51)、39.2％(20/51)、45.1％(23/51)；丢失率分别是21.6 ％(11/51)、33.3％(17/51)、27.5％(14/51).HLA-ABC在喉鳞癌组织中的表达分别与TAP-1(r=0.460,P＜0.05)、LMP-7(r=0.685,P＜0.05)的表达呈正相关.在喉鳞癌组织中HLA-ABC、TAP-1、LMP-7的表达均与肿瘤T分期相关(x2值分别为8.61、9.72、8.97,P值均＜0.05),TAP-1、LMP-7的表达分别与肿瘤TNM分期相关(x2值分别为9.18和7.70,P值均＜0.05).单因素分析显示,HLA-ABC阴性表达组与表达下调组、阳性表达组间患者5年累积生存率(cumulative survival rate,CS)及3年无瘤生存率( disease-free survival,DFS)差异有统计学意义(CS组间比较X2值分别为16.18和15.96,DFS组间比较x2值分别为12.54和6.31,P值均＜0.05)；TAP-1阳性表达组与表达下调组、阴性表达组向5年 CS及3年DFS差异有统计学意义(CS组间比较x2值分别为4.23和4.42,DFS组间比较x2值分别为4.12和4.56,P值均＜0.05)；LMP-7阴性表达组与表达下调组、阳性表达组患者5年CS及3年DFS差异有统计学意义(CS组间比较x2值分别为11.46和14.58,DFS组间比较x2值分别为14.17和8.74,P值均＜0.05).多因素分析提示,颈淋巴结转移、复发情况和HLA-ABC表达水平是影响本组喉鳞癌患者预后的重要危险因素(P值均＜ 0.05).结论 喉鳞癌组织中HLA-ABC、TAP-1、LMP-7阴性表达与表达下调不利于肿瘤抗原的递呈及细胞毒性T淋巴细胞向肿瘤实质浸润与杀伤.HLA-Ⅰ类分子、TAP-1、LMP-7阴性表达与表达下调可能促进了癌细胞的发生、侵袭及发展,可能是肿瘤逃避机体免疫监视的方式之一.     

高迁移率族蛋白1在喉鳞状细胞癌组织及血清中的表达及意义

目的 研究高迁移率族蛋白1( high mobility group box-1,HMGB1) mRNA及蛋白在喉鳞状细胞癌(简称喉鳞癌)组织及患者血清中的表达及其临床意义.方法 反转录聚合酶链反应( RTPCR)及Western blot分别检测HMGB1 mRNA及蛋白在30例喉鳞癌患者肿瘤组织及癌旁黏膜组织中的表达;酶联免疫吸附试验( ELISA)检测HMGB1蛋白在喉鳞癌患者及正常志愿者血清中的表达.结果RT-PCR结果示HMGB1 mRNA(HMGB1/GAPDH)在喉鳞癌组织、癌旁黏膜组织中的相对表达量((x)±s)分别为1.25±0.12、0.32±0.04,差异有统计学意义(t=40.27,P＜0.05);Western blot结果示HMGB1蛋白(HMGB1/β-actin)在喉鳞癌和癌旁黏膜组织中的相对表达量分别为1.29 ±0.10、0.34±0.03,两组间差异有统计学意义(t=49.84,P＜0.05).结果显示HMGB1 mRNA和蛋白的表达水平与患者的T分级、临床分期、有无淋巴转移及是否吸烟有关(P值均＜0.05);而与年龄、性别、是否饮酒及肿瘤病理分级无关(P＞0.05).ELISA结果显示,喉鳞癌患者及正常志愿者血清中HMGB1质量浓度分别为(24.80±14.08) ng/ml、(23.58±14.69)ng/ml,差异无统计学意义(t=0.37,P＞0.05).结论 HMGB1 mRNA及蛋白在喉鳞癌组织中的表达显著高于癌旁黏膜组织,且表达水平与T分级、临床分期及有无淋巴转移有关.     

组织缺氧诱导因子-1α及相关靶基因在喉鳞状细胞癌组织中的表达

目的 检测喉鳞状细胞癌(简称鳞癌)组织缺氧诱导因子-10α( hypoxia inducible factor 1 alpha,HIF-1α)、葡萄糖转运蛋白-1(glucose transporter protein-1,GLUT-1)、血管内皮生长因子( vascular endothelial growth factor,VEGF)的蛋白表达情况,并通过体外实验进一步验证低氧对喉鳞癌细胞HIF-1α及其下游靶基因GLUT-1、VEGF的表达上调方面的重要促进作用.方法 采用SP免疫组织化学和免疫细胞化学方法检测喉鳞癌组织和Hep-2细胞中HIF-1α、GLUT-1、VEGF蛋白的表达,分析HIF-1α与GLUT-1、VEGF表达的相关性.结果 35例喉癌患者组织切片中16例HIF-1α表达阳性(45.7％),16例GLUT-1表达阳性(阳性率45.7％),19例VEGF表达阳性(阳性率54.3％).HIF-1α和VEGF的表达水平与喉癌病理分级和淋巴转移有关(P值均＜0.05);GLUT-1表达水平与淋巴转移有关(P＜0.05).体外实验结果显示,Hep-2细胞在低氧条件下HIF-1α、GLUT-1、VEGF的蛋白表达明显增加(P值均＜0.05).结论 HIF-1α可作为转录调控因子参与调控GLUT-1、VEGF的表达,进而在喉癌的发生、侵袭、转移过程中发挥重要作用.     

Galectin-3和血管内皮生长因子在喉鳞状细胞癌组织中的表达

目的:探讨喉鳞状细胞癌组织中Galectin-3和血管内皮生长因子(VEGF)蛋白表达及其在肿瘤分化,生长及转移中的相关意义.方法:应用免疫组织化学SP法和免疫印记法分别对29例喉鳞状细胞癌、18例喉良性病变组织中Galectin-3和VEGF蛋白表达水平进行了检测.结果:Galectin-3(89.7%)和VEGF(86.2%)在喉鳞状细胞癌组的阳性表达率与正常对照组比较均差异有统计学意义(均P＜0.05).喉高分化鳞状细胞癌组织中Galectin-3的表达与低分化鳞状细胞癌相比差异有统计学意义(P＜0.05);并凡,Galectin-3与VEGF在喉鳞状细胞癌组织中的表达呈正相关关系(r=0.423,P＜0.05).结论:喉鳞状细胞癌组织中Galectin-3和VEGF的高表达可能对肿瘤的组织分化和转移起重要作用.     

Epidemiology of basal cell carcinoma and squamous cell carcinoma of the pinna

This is a retrospective study designed to compare the incidence of basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) in the head and neck skin area with special reference to the pinna. The results showed 426 patients had 460 cutaneous malignancies in the head and neck area, managed by four specialties (ENT, Dermatology, General Surgery and GPs) over the period 1994-99. The lesions comprised 375 (80.47 per cent) BCC and 85 (18.47 per cent) SCC. In cases of BCC the facial areas were commonly involved (88 per cent), whilst SCC was almost equally distributed between the most (face, forehead and nose) and least exposed areas (pinna and scalp). The overall ratio of BCC to SCC remained four to one in the head and neck area. In 41 patients with 51 lesions over the pinna there were 29 (56.8 per cent) BCC and 22 (43.1 per cent) SCC hence the ratio was 1.3 to 1 for this site. We conclude that in the case of a suspicious lesion over the pinna, the risk of SCC is comparatively much higher. With increasing awareness of early and quick diagnosis of cancer cases, it is recommended that these patients should be referred urgently to prevent the significant morbidity associated with invasive SCC.     

Primary composite squamous cell carcinoma and large cell neuroendocrine carcinoma of the hypopharynx

Neuroendocrine carcinomas are rare neoplasms of the larynx and hypopharynx. Tumours composed of both neuroendocrine and squamous cell elements are very rare. We report a case of a primary hypopharyngeal carcinoma composed of both squamous cell and large cell neuroendocrine carcinoma and discuss the treatment of this patient and management of neuroendocrine carcinomas of the larynx and hypopharynx.     

Merkel cell carcinoma

PURPOSE: To determine the natural history and treatment outcomes for patients with Merkel cell carcinoma. METHODS: Review of the literature. RESULTS: The probability of regional node involvement at presentation exceeds 50%; few patients present with distant metastases. Comprehensive treatment of the primary site and regional lymphatics with surgery or radiotherapy results in the highest likelihood of cure. The role of adjuvant chemotherapy remains investigational. CONCLUSION: The probability of regional dissemination at diagnosis is high. The optimal treatment is resection of the primary tumor and treatment of the regional lymphatics. Resection of all gross tumor should be accomplished followed by local-regional radiotherapy in most patients.     

Papillary squamous cell carcinoma versus verrucous squamous cell carcinoma of the head and neck

Papillary squamous cell carcinoma and verrucous squamous cell carcinoma of the head and neck may be confused. The clinicopathological profile of the two neoplasms is presented and the differential diagnosis is discussed. A correct diagnosis is imperative in order to institute the most appropriate treatment.     

Prognoses of oral basaloid squamous cell carcinoma and squamous cell carcinoma: a comparison

OBJECTIVE: To compare clinical and prognostic features in patients with basaloid squamous cell carcinoma (BSCC), poorly differentiated squamous cell carcinoma (PDSCC), and well to moderately differentiated squamous cell carcinoma (W/MSCC) of the oral cavity. DESIGN: Retrospective cohort. SETTING: Referral tertiary center. PATIENTS: Seventeen patients with primary oral BSCC, 27 with PDSCC, and 27 with SCC. INTERVENTION: The 71 patients all had surgery and 52 had postoperative radiotherapy. MAIN OUTCOME MEASURES: Recurrences and survival. RESULTS: The median follow-up time was 52.4 months for patients with BSCC, 22.2 months for those with PDSCC, and 13.8 months for those with SCC. No statistically significant differences on survival were found among the BSCC, PDSCC, and SCC groups. The 5-year cancer-specific survival rates were 50% for patients with BSCC, 37% for those with PSCC, and 49% for those with W/MSCC (P =.71); the 5-year overall survival rates were 46% for patients with BSSC, 18% for those with PSCC, and 41% for those with W/MSCC (P =.25). Disease-free survival was not significantly different among the BSCC, PSCC, and W/MSCC groups (P =.57). The 5-year rate of disease-free survival was 40% for patients with BSCC, 37% for those with PSCC, and 53% for those with W/MSCC. CONCLUSIONS: The clinical course of BSCC is similar to the courses of PSCC and W/MSCC when clinical T and N classifications are matched. Prognosis does not differ for patients with BSCC of the oral cavity and those with conventional oral squamous cell carcinomas PSCC and W/MSCC.     

Small cell epidermoid carcinoma of salivary glands. 'Pseudo'-oat cell carcinoma

Two patients had small cell carcinomas of the salivary glands, with pathological features indicating squamous differentiation, heretofore not described. One is free of disease at seven years, and the second is alive, with regional metastases at four years. Sections from one tumor were studied by electron microscopy and revealed tonofilaments and desmosomes. Most cases of small cell carcinomas of the salivary glands have been considered akin to bronchogenic oat cell carcinoma. Their less aggressive behavior, however, suggests that at least some of these tumors were not true oat cell carcinomas. Our findings, and those of others, indicate that small cell carcinomas of the salivary glands (or head and neck) represent a heterogeneous group. Electron microscopy should be used to determine the exact nature of these neoplasms. If an oat cell nature is ruled out, local and regional treatment should be aggressive, since small cell carcinomas other than oat cell appear not to have a dismal prognosis.     

Invasive squamous cell carcinoma within verrucous carcinoma

A case of invasive squamous cell carcinoma within a verrucous carcinoma is presented in order to illustrate the potential problem of underdiagnosis of these lesions. The epidemiology, natural history and histopathology of verrucous carcinoma, and features which distinguish it from invasive squamous cell carcinoma, are reviewed. Unless rigorous attention is paid to histologic detail, a focus of invasive squamous cell carcinoma may escape detection and radiation-induced anaplastic transformation may later be suspected.