听神经瘤S518磷酸化Merlin蛋白的表达及与CD44结合特性

目的 探讨听神经瘤组织中S518磷酸化对Merlin蛋白与细胞表面糖蛋白CD44结合能力的影响及其与肿瘤生长的关系.方法 35例听神经瘤组织标本均经过病理学证实,用免疫组织化学技术和免疫印迹技术分析S518磷酸化Merlin蛋白的表达、组织学分布情况,以手术中切除的正常颅神经作对照.将各标本中S518磷酸化Merlin条带的相对灰度值与听神经瘤临床分期、瘤体大小、病程及囊性变进行相关分析.用免疫沉淀技术比较野生型Merlin蛋白及S518磷酸化Merlin蛋白与CD44结合能力的差异.结果 在听神经瘤组织中Merlin蛋白发生S518磷酸化且分布集中于核周;正常对照神经组织中Merlin也发生S518磷酸化,但是其含量较听神经瘤组织低.S518磷酸化的Merlin蛋白在听神经瘤组织中的表达量与临床分期、肿瘤大小、病程及囊性变之间无统计学相关关系.在听神经瘤组织中,与S518磷酸化的Merlin蛋白结合的CD44量高于与野生型Merlin蛋白结合的CD44的量.结论 在听神经瘤组织中,S518磷酸化的Merlin蛋白与CD44的结合能力增加,CD44与野生型Merlin蛋白和S518磷酸化Merlin蛋白结合后可能产生不同细胞生物学行为.     

程序性细胞死亡因子5在喉鳞状细胞癌组织中的表达及临床意义

目的 探讨程序性细胞死亡因子5（PDCD5）在喉鳞状细胞癌组织中的表达及与临床病理指标的相关性。方法 采用逆转录多聚酶链式反应（RT PCR）技术和Western blot免疫印记技术检测41例喉鳞状细胞癌及29例癌旁正常黏膜组织中PDCD5mRNA和蛋白的表达情况,进一步应用免疫组化方法验证其在喉鳞状细胞癌组织中的定位表达。结果 PDCD5 mRNA喉鳞状细胞癌组织中较癌旁正常组织的表达明显降低,表达差异有统计学意义（P=0.006）;Western blot结果显示PDCD5 蛋白在喉鳞状细胞癌组织中较癌旁正常组织的表达明显降低,表达差异有统计学意义（P=0.012）。肿瘤组织中PDCD5 mRNA表达缺失与PDCD5蛋白表达缺失之间存在明显的相关性（Pearson r= 0.695, P<0.01）。免疫组化进一步证实PDCD5在肿瘤组织中的高频率缺失。χ2检验分析发现PDCD5蛋白表达缺失与喉鳞状细胞癌患者的年龄、性别、肿瘤原发部位和有无淋巴结转移无相关性,但是与肿瘤TNM分期以及组织学分级之间相关(mRNA P=0.032, P=0.021;蛋白 P=0.027, P=0.016)。结论 PDCD5在喉癌中多呈低表达,在喉鳞状细胞癌的发生、发展过程中可能起重要作用。     

CPS1在喉鳞癌组织中的表达及临床意义

目的 研究氨基甲酰磷酸合成酶1(CPS1)在喉鳞癌患者组织中的表达水平及与临床病理指标的相关性,探讨CPS1对喉鳞癌的临床意义。 方法 采用逆转录实时定量聚合酶链反应(RT-qPCR)及Western blot免疫印迹技术,分别检测喉鳞癌组织及癌旁组织中CPS1 mRNA及蛋白表达水平,同时采用免疫组织化学法检测 CPS1在喉鳞癌组织中的表达水平及其结构中的分布情况。 结果 RT-qPCR 和Western blot结果显示,与癌旁组织相比,喉鳞癌组织CPS1 mRNA 和蛋白表达明显增高,表达差异有统计学意义(P<0.01);免疫组化结果显示CPS1表达与肿瘤分化程度、临床分期、淋巴结转移等因素密切相关(P<0.05),而与患者年龄、性别、吸烟史、饮酒史及肿瘤部位无关(P>0.05)。 结论 在喉鳞癌组织中CPS1的mRNA和蛋白表达明显升高,且与临床进展密切相关,说明CPS1在喉癌发生及转移过程中起到一定的作用,可能作为喉癌诊断的重要指标。     

听神经瘤临床特征与肿瘤细胞增殖因子的相关性研究

目的 探讨肿瘤增殖标记指数(1abeling index,LI)Ki-67、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、转移生长因子-β1(transforming growth factor-β1,TGF-β1)在听神经瘤组织标本中表达的意义及其与临床行为之间的相关性.方法 回顾性分析听神经瘤手术患者53例的临床资料,对其石蜡切片病理标本进行Ki-67、PCNA和TGF-β1免疫组化学常规步骤染色.用SAS 9.0统计软件包进行统计分析.结果 53例中,Ki-67阳性率77.4%(41/53),PCNA阳性率84.9%(45/53);Ki-67阳性与病程和临床生长率有关(t=2.14和2.70,P值均<0.05);PCNA阳性与临床生长率有关(t'=-4.45,P<0.05);TGF-β1阳性表达83.0%(44/53),与Ki-67增殖标记指数呈正相关(r=0.36,P<0.05).53例中囊性变者比非囊变者肿瘤直径明显增大(Z=4.44,P<0.05);囊性变者与非囊变者,其肿瘤增殖标记指数Ki-67、PCNA、TGF-β1和病程之间无相关性.听神经瘤囊性变者比非囊变者临床生长率增大,但差异无统计学意义.结论 Ki-67和PCNA与肿瘤的细胞增殖有关,增殖指数与临床生长率有关.双向调节因子TGF-β1参与了听神经瘤的生物学行为.囊性变听神经瘤,其增长与肿瘤的增殖活性关系不大,其增长是囊的容积增加,而非肿瘤细胞生长速度的加快.     

髓鞘结构在听神经瘤组织中的检测

目的:探讨来源于施万细胞的人类听神经细胞瘤组织中是否存在髓鞘结构及其细胞学特点。方法:应用免疫荧光标记、Western blotting及电镜等方法检测听神经瘤组织中的髓鞘结构及重要髓鞘蛋白。结果:电镜结果发现:听神经瘤组织中虽然缺失成熟的轴索结构但可以检测到早期的髓鞘样结构。免疫荧光研究和Western blotting研究结果显示:听神经瘤组织中检测不到轴索的标志性抗体——神经丝200的表达。并且启动髓鞘化的前哨信号及髓鞘致密化的重要组成蛋白——髓鞘碱性蛋白在听神经瘤组织中也无法检测到。而一种非成熟施万细胞的标志抗体——神经生长因子受体p75,在听神经瘤组织中可以检测到其表达。结论:在听神经瘤组织中的施万细胞失去了完成髓鞘化进程和重新形成致密的髓鞘并包绕轴索的能力,处于前髓鞘化施万细胞阶段。     

FoxM1在喉鳞状细胞癌中的表达及其临床意义

目的:研究FoxM1在喉鳞状细胞癌组织中的表达及其临床意义。方法:运用免疫组织化学方法检测89例喉鳞状细胞癌组织、89例癌旁组织及20例喉乳头状瘤组织中FoxM1蛋白的表达情况,分析其与喉鳞状细胞癌临床病理参数的关系。采用反转录-聚合酶链反应(RT-PCR)检测20例喉鳞状细胞癌、20例喉乳头状瘤组织以及20例癌旁组织中FoxM1mRNA表达情况。结果:FoxM1mRNA和蛋白表达的阳性率在喉鳞状细胞癌、喉乳头状瘤及癌旁组织中逐渐降低,其差异有统计学意义(P<0.05)。FoxM1蛋白的表达随组织分化程度增高而降低(P<0.05);随T分期增高而增加(P<0.05);有淋巴结转移组高于无淋巴结转移组(P<0.05),随临床分期的增加而表达增加(P<0.05)。FoxM1表达率与患者年龄、性别、癌原发部位无相关性(P>0.05)。结论:FoxM1蛋白在喉鳞状细胞癌组织中表达的上调可能与喉鳞状细胞癌的发生及浸润转移有关。     

上皮生长因子受体-信号转导与转录激活因子3信号转导通路与喉乳头状瘤的相关性研究

目的：探讨喉乳头状瘤中上皮生长因子受体（EGFR）和信号转导与转录激活因子3（STAT3）的表达与其生物学行为的关系。方法：采用RT-PCR、免疫组织化学方法和Western blot,检测42例喉乳头状瘤和15例正常喉黏膜组织中EGFR和STAT3、磷酸化-STAT3（p—STAT3）的mRNA和蛋白质表达情况,同时将蛋白质表达水平与喉乳头状瘤的临床病理参数进行相关性分析。结果：EGFR与STAT3的mRNA在喉乳头状瘤组织表达水平显著高于正常喉组织（P〈0．05,P〈0．01）。免疫组织化学及Western blot方法也证实喉乳头状瘤组织中EGFR与STAT3（p-STAT3）的蛋白质表达水平显著高于正常喉组织（P〈0．05）。EGFR与sTAT3（P—STAT3）蛋白质在喉乳头状瘤组织的表达显著关联（P〈0．05）,STAT3蛋白与喉乳头状瘤的复发和恶变有明显相关性（P〈0．05）。结论：EGFR—STAT3信号转导通路参与喉乳头状瘤的发生,STAT3蛋白的持续活化可能促进喉乳头状瘤的复发与恶变。     

喉鳞状细胞癌Paxillin表达的临床意义

目的 探讨喉鳞状细胞癌Paxillin蛋白表达的临床意义。方法 运用免疫组化S-P法检测石蜡包埋喉鳞癌标本74例，以及癌旁正常组织70例，癌旁轻度、中度、重度不典型增生黏膜上皮组织65、69、72例，分析比较Paxillin蛋白表达水平与患者各类临床资料的相关性及其临床意义。结果 喉鳞癌组织Paxillin阳性率明显高于正常组织标本（P〈0．05），不同分化程度癌组织的Paxillin阳性率也存在显著性差异，低分化者阳性率明显高于高和中分化组（P=0．001），伴有颈部淋巴结转移者阳性率明显高于无转移者（P=0．000），而且临床Ⅳ期病例阳性率又明显高于其他各期患者（P=0．000）。结论 Paxillin的阳性表达水平与喉鳞状细胞癌的分化程度、颈淋巴结转移、临床分期有明显相关性，对其病理过程具有一定影响。     

BUB1在喉鳞状细胞癌中的表达及临床意义

目的：探讨喉鳞状细胞癌（LSCC）组织中BUB1蛋白的表达及临床意义。方法：采用免疫组织化学EliVision二步法检测55例LSCC组织和30例癌旁正常黏膜组织中BUB1蛋白的表达,并分析其与LSCC临床病理特征的关系。结果：BUB1蛋白在LSCC组织中的阳性表达率为50．9％（28／55）,低于癌旁正常黏膜组织83．3%（25／30）,两者比较差异有统计学意义（P〈0．01）;BUBl蛋白的表达与LSCC分化程度及淋巴结转移有相关性（均P〈0．05）,而与患者性别、年龄、吸烟、肿瘤原发部位、T分期及临床分期无相关性（均P〉0．05）。结论：BUBl蛋白的低表达与LSCC的发生、发展有关,BUB1可能作为预测LSCC转移及预后的生物学指标之一。     

TrKB与VEGF在鼻咽癌组织中的表达及临床意义

目的探讨血管内皮生长因子(VEGF)、酪氨酸激酶B(TrKB)在鼻咽癌(NPC)组织中的表达并比较两者表达的相关性及分析与肿瘤临床特征的关系。方法采用免疫组化方法检测经活检确诊为NPC的患者55例及鼻咽炎组织30例中VEGF、TrKB蛋白表达水平,比较二两者表达差异。根据患者年龄和性别、肿瘤临床分型、原发灶大小、淋巴结转移和有无远处转移将病例分组,应用双变量相关Pearson检验分析研究VEGF、TrKB表达与肿瘤临床病理特征之间的关系。结果①VEGF蛋白于肿瘤细胞浆及血管内皮细胞浆中均表达,TrKB蛋白阳性表达绝大多数位于细胞浆内。在55例NPC患者中,45例TrKB阳性表达,阳性表达率为81.8%;47例VEGF阳性表达,阳性表达率为85.4%。30例鼻咽炎中,只有3例TrKB阳性表达,阳性表达率为10.0%;5例VEGF阳性表达,阳性表达率为16.7%。肿瘤组织中TrKB、VEGF阳性表达率明显高于鼻咽炎组织（P<0.05）,NPC组织中VEGF与TrKB表达有明显相关性（R=0.716,P<0.01）;②VEGF蛋白表达与NPC的临床分期(P=0.002)、肿瘤的直径大小(P=0.009)及有无淋巴结转移(P=0.001)明显相关,与患者性别、年龄、有无远处转移无关（P＞0.05）。TrKB蛋白表达与临床分期(P=0.00)、肿瘤的直径大小(P=0.00)、有无淋巴结转移（P=0.006）明显相关,与患者性别、年龄、有无远处转移无关（P＞0.05）。结论VEGF、TrKB在NPC组织中表达增加,可能与NPC的发生发展有关。TrKB可能与NPC血管形成有关。 （     

Lipid raft localization of ErbB2 in vestibular schwannoma and schwann cells.

HYPOTHESIS: ErbB2 resides in lipid rafts (regions of receptor regulation) in vestibular schwannoma (VS) cells. BACKGROUND: ErbB2 is a growth factor receptor critical for Schwann cell (SC) proliferation and development. ErbB2 localization and activity may be regulated by merlin, an adaptor protein deficient in VS. Lipid rafts are microdomains in the plasma membrane that amplify and regulate receptor signaling. Persistence of erbB2 in lipid rafts in VS due to merlin deficiency may explain increased VS cell growth. METHODS: Protein extracts from VS or rat sciatic nerve (proximal or distal to a crush injury) were isolated into lipid raft and nonraft fractions and immunoblotted for erbB2, phosphorylated erbB2, and merlin (for sciatic nerve). Cultured VS cells were probed with anti-erbB2 antibody and a lipid raft marker, cholera toxin B (CTB). RESULTS: ErbB2 moves to lipid rafts in proliferating SCs and is persistently localized to lipid rafts in VS cells. ErbB2 is phosphorylated (activated) in lipid rafts. ErbB2 colocalized with CTB in cultured VS cells, confirming raft targeting. Merlin also persistently localized to lipid rafts in SCs, and its relative phosphorylation increased in proliferating cells. CONCLUSION: Lipid raft localization of erbB2 in proliferating SCs and in VS cells supports a critical role for lipid rafts in amplifying/regulating erbB2 signaling. Merlin resides in lipid rafts in SCs, and its phosphorylation increases in proliferating SCs, suggesting it regulates cell proliferation within lipid rafts. The absence of merlin in VS may therefore lead to persistent erbB2 localization to lipid rafts and increased cell proliferation.     

Co-expression of transforming growth factor-beta1 and glial cell line-derived neurotrophic factor in vestibular schwannoma.

HYPOTHESIS: Transforming growth factor-beta1, glial cell line-derived neurotrophic factor, and their receptors are expressed in vestibular schwannoma, and the expression data correlate with the proliferation activity (Ki-67 labeling index) and the clinical growth rate of vestibular schwannoma tissue. BACKGROUND: Glial cell line-derived neurotrophic factor is a potent growth factor for the central and peripheral nervous system. Recent results demonstrate that glial cell line-derived neurotrophic factor requires transforming growth factor-beta to exert its trophic effect on neural tissue. A functional role, including that in Schwann cell proliferation, is discussed for both transforming growth factor-beta1 and glial cell line-derived neurotrophic factor. METHODS: Immunohistochemical analysis for transforming growth factor-beta1 and glial cell line-derived neurotrophic factor and their receptors TbetaR II, GFRalpha-1, and Ret was performed on formalin-fixed, paraffin-embedded archival surgical specimens. The Ki-67 labeling index (mean Ki-67 labeling index and highest Ki-67 labeling index for Antoni Type A and Type B regions) and the clinical growth rate of vestibular schwannoma were determined and correlated with the expression patterns of the examined neurotrophic factors and their receptors. RESULTS: Results demonstrate co-expression of transforming growth factor-beta1 and glial cell line-derived neurotrophic factor with higher levels in Antoni Type A than in Antoni Type B regions. Ninety-five percent of vestibular schwannomas exhibited transforming growth factor-beta1 immunoreactivity, and glial cell line-derived neurotrophic factor expression was found in 100% of vestibular schwannoma specimens. Fifty percent of vestibular schwannoma displayed TbetaR II immunostaining, 100% showed positive reactions for GFRalpha-1, and 86% showed positive reactions for Ret. Statistical analysis revealed no significant correlation in neurotrophin expression related to sex, age, tumor size, clinical growth rate, or Ki-67-labeling indices. CONCLUSIONS: Expression of transforming growth factor-beta1 and glial cell line-derived neurotrophic factor may suggest a biological role for both growth factors in vestibular schwannomas. Trophic transforming growth factor-beta/glial cell line-derived neurotrophic factor synergism seems possible and is underscored by co-expression of both neurotrophic factors and their receptors.     

听神经瘤的分子生物学研究进展

听神经瘤由于它特殊的自然病程而引起人们的重视。近10年来有关听神经瘤发病机制及其自然进程方面的分子生物学研究发现,在家族性听神经瘤病人中其基因型和表显型有较明确的关系,而散发的听神经瘤中关系目前尚未完全清楚。NF2基因的失活只在肿瘤发生的早期起作用,可能还有其他基因参与肿瘤的分化调节,进一步的研究是必要的。     

Postoperative quality of life in vestibular schwannoma patients measured by the SF36 Health Questionnaire

OBJECTIVE: To quantify the postoperative quality of life in patients following surgical treatment for vestibular schwannoma. STUDY DESIGN: Patient self-assessment using the short form 36 (SF36) multidimensional quality of life health questionnaire. Sex- and age-matched normalized scores were calculated using a standardized process and accepted normative data. SETTING: Tertiary referral skull base unit. RESULTS: An 80% response rate (90 patients) was achieved. The postoperative quality of life in vestibular schwannoma patients, as quantified by seven of the eight SF36 health scales was less than the appropriate matched healthy standard. Comparison of a variety of preoperative patients and tumor factors-different operative approaches (translabyrinthine and retrosigmoid), tumor size (group cut of points of tumor diameter 1.5 mm and 2.5 mm), patient sex, and ranking of patient age-showed no statistically significant difference in measured quality of life outcomes for each of these traditional predictors. CONCLUSION: Reduced quality of life in patients after surgical treatment for vestibular schwannoma, coupled with the low tumor growth rates and minimal preoperative symptoms, supports a conservative approach to patient management. The advantages and disadvantages of a variety of approaches used to measure the quality of life after surgical treatment of vestibular schwannoma and their impact on clinical decision making for patients, are discussed.     

Assessment of real-time clinical facial function during vestibular schwannoma resection

OBJECTIVES: To describe a technique for quantifying residual facial function after vestibular schwannoma surgery. The intraoperative electrophysiological results are correlated with immediate postoperative clinical facial function to assess technique validity. STUDY DESIGN: Prospective blinded study. METHODS: Thirty-two patients undergoing translabyrinthine resection of vestibular schwannoma were included. Compound muscle action potential (CMAP) amplitude was calculated to supramaximal stimulation of the facial nerve, proximal to vestibular schwannoma compression, at the brain stem. The procedure was repeated after tumor removal. Comparison of the two CMAP amplitudes enabled estimation of change in facial function during surgery. The data were correlated with intracranial tumor diameter and immediate postoperative clinical facial function. RESULTS: CMAP amplitude recorded after tumor resection correlated with immediate postoperative clinical facial function (0.879, P < .01). Correlation was improved when residual facial function was calculated (0.944, P < .01). In contrast, preoperative tumor size had relatively poor correlation with immediate postoperative facial function (0.688, P < .01). CONCLUSIONS: Comparison of electrophysiological data before and after tumor removal reduces intersubject variability resulting from intersubject variation in facial muscle morphology. Residual facial function closely correlates with immediate postoperative clinical facial function, assessed using the House-Brackmann grading system. From the data presented, the technique can inform the surgeon of current clinical facial function at any point during the dissection process.     

听神经瘤术后单侧耳聋听力重建问题初探

摘要：目的探讨听神经瘤术后单侧耳聋患者人工听觉重建的效果。方法回顾性分析2016年7~12月上海交通大学医学院附属第九人民医院耳鼻咽喉头颈外科收治的听神经瘤术后单侧耳聋行人工听觉植入患者8例，其中同侧一期骨桥植入患者6例，二期同侧骨桥植入2例。比较术前及术后1年噪声下言语识别能力以及生活质量改变。结果所有病例均成功手术，肿瘤均完全切除。至随访1年，8例患者均正常使用听觉植入装置。骨桥植入患者噪声下言语识别能力明显提高，生活质量明显改善。结论骨桥可以帮助听神经瘤术后单侧耳聋患者有效地进行听觉功能重建，需要根据患者情况制订个体化的治疗方案。

     

Neurotrophic factor expression in vestibular schwannoma. An overview

The vestibular schwannoma is a benign, slow-growing neoplasm that originates from the neurolemmal sheath of the vestibular branch of the VIIIth cranial nerve. This tumor entity accounts for 6 % of all intracranial tumors and the annual incidence of newly diagnosed vestibular schwannoma is reported as 13 per million. The molecular pathogenesis of both sporadic vestibular schwannoma and those occurring in neurofibromatosis type II appears to be associated with an aberration of a tumor suppressor gene on chromosome 22q12. The biological background for the various growth patterns of vestibular schwannoma is, however, largely unknown. This differing clinical and biological behaviour of vestibular schwannoma may be explained by the presence of neurotrophic factors. The results of recent immunohistochemical studies demonstrate the co-expression of transforming growth factor (TGF)-beta 1 and glial cell line-derived neurotrophic factor (GDNF) in vestibular schwannoma and suggest a trophic synergism of both neurotrophic factors in this tumor. Moreover, expression of numerous different neurotrophic factors has been shown in studies of nerve growth factor (NGF), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), fibroblast growth factor (FGF), neuregulin (NRG) and erythropoietin (EPO) indicating a biological role in development, maintainance or growth of vestibular schwannoma. In this article, we summarize the findings on neurotrophic factor expression and discuss their characteristics and biological role in vestibular schwannoma.