p-ERK5在快速衰老小鼠耳蜗血管纹中的增龄性变化

目的探讨快速衰老小鼠耳蜗血管纹中磷酸化的细胞外信号调节激酶5（phosphorylated extracellular signal regulated kinase 5, p ERK5）表达的增龄性变化。方法选用3、5、7月龄的快速衰老小鼠亚系８（senescence accelerated mouse, ＳＡＭＰ８）各６只,分别进行８ｋＨｚ短纯音听性脑干反应（ＡＢＲ）检测,并用免疫组化染色方法分别检测各月龄组小鼠耳蜗血管纹细胞中p ERK5的表达,分析其增龄性变化。结果3、５、７月龄小鼠耳蜗血管纹细胞中p ERK5平均光密度值分别为0.3838±0.0020、0.3646±0.0010、0.3423±0.0036; p ERK5在不同月龄快速衰老小鼠耳蜗组织中的表达光密度值随着月龄增加而显著降低（P＜０.０５）。结论随着年龄相关性功能减退,快速衰老小鼠耳蜗血管纹中p ERK5的表达水平逐渐降低,推测p ERK5可能与维持正常的耳蜗功能及听觉有关。     

miR-485-5p在鼻咽癌组织中的表达及临床意义

目的探讨miR-485-5p在鼻咽癌组织中的表达及临床意义。方法采用qRT-PCR检测76例鼻咽癌和28例鼻咽部黏膜组织中miR-485-5p的表达,并结合临床信息,统计分析其表达水平与鼻咽癌患者临床病理参数的相关性。结果miR-485-5p在鼻咽癌组织中的表达较鼻咽部黏膜组织显著下降（P<0.05）;同时miR-485-5p低表达与鼻咽癌患者的淋巴结转移密切相关（P=0.001）,而与鼻咽癌患者的年龄、性别、T分级和TNM分期无明显相关（P> 0.05）。结论本研究表明miR-485-5p为转移相关基因,可能在鼻咽癌转移中发挥了重要作用。     

MicroRNA-34c-5p在喉癌前病变组织中的特征性差异表达及临床病理分析

目的　探讨microRNA-34c-5p（miR-34c）在喉癌前病变组织中差异表达情况,并进行临床病理相关性分析。方法　按照2005年世界卫生组织（WHO）病理诊断标准,94例喉癌前病变分为轻度异型增生、中度异型增生、重度异型增生及原位癌;实时定量聚合酶链式反应（quantitative real-time polymerase chain reaction,qRTPCR） 方法检测94例喉癌前病变组织中miR-34c在各组间的差异表达情况,并结合临床资料及随访结果分析miR-34c与喉癌前病变病例临床因素的相关性。结果　94例喉癌前病变病例年龄37~86岁,平均58岁,男性85例（90.4%）,女性9例（9.6%）。轻度异型增生组20例（21.3%）,中度异型增生组23例（24.5%）,重度异型增生组26例（27.7%）,原位癌组25例（26.6%）。随喉癌前病变级别增高,miR-34c在喉癌前病变组织中呈上调表达趋势,总体组间存在统计学差异（F =10.182,P =0.00）,而在浸润性鳞状细胞癌组下降,甚至明显低于正常鳞状上皮组织的表达均值。临床病理分析显示饮酒病例中表达明显高于非饮酒病例,且miR-34c与喉癌前病变患者饮酒因素存在明显线性依存关系（r =0.31,P =0.03）。结论　MiR-34c在喉癌前病变组织中的上调表达趋势,可能为辅助喉癌前病变病理分级和早期喉癌诊断提供一定的分子病理参考价值,miR-34c的上调表达与喉癌前病变饮酒因素呈线性相关,对于指导喉癌前病变病例的临床管理可能具有重要临床意义。     

miR-30-5p通过下调FOXG1表达抑制视网膜母细胞瘤细胞增殖

目的 探讨miR-30-5p对视网膜母细胞瘤细胞增殖的影响及作用机制。 方法 采用实时荧光定量聚合酶链式反应(qRT-PCR)检测miR-30-5p在视网膜母细胞瘤细胞系和人视网膜内皮细胞(HRECs)中的表达。利用TargetScan数据库进行生物信息学分析并预测miR-30-5p靶基因,双荧光素酶报告基因实验验证miR-30-5p与FOXG1的3'UTR结合能力及靶向关系,qRT-PCR和Western blotting分别检测miR-30-5p对FOXG1的mRNA与蛋白表达影响。瞬时转染Y79细胞后,通过CCK8法检测各组Y79细胞的增殖。 结果 与HRECs比较,miR-30-5p在视网膜母细胞瘤细胞中表达降低,FOXG1在视网膜母细胞瘤细胞中表达升高。生物信息学预测结果显示miR-30-5p与FOXG1存在结合位点,qRT-PCR和Western blotting显示miR-30-5p可负调控FOXG1的mRNA和蛋白的表达,双荧光素酶实验结果证实miR-30-5p与FOXG1 3'UTR存在靶向关系。瞬时转染miR-30-5p可升高Y79细胞中miR-30-5p的表达水平,抑制Y79细胞的增殖活力。过表达FOXG1可逆转miR-30-5p上调对Y79细胞增殖的影响。 结论 miR-30-5p通过下调FOXG1表达抑制视网膜母细胞瘤Y79细胞的增殖。     

Correlation between proliferating cell nuclear antigen and p53 protein expression and 5-year survival rate in nasopharyngeal carcinoma

PURPOSE: To assess the prognostic significance of p53 protein and proliferating cell nuclear antigen (PCNA) expression in nasopharyngeal carcinoma. MATERIALS AND METHODS: This study included 79 patients who had received treatment and regular follow-up for at least 5 years at a single institute. We used immunohistochemistry staining to assess p53 protein expression and PCNA labeling index (LI). Analyses were conducted on the association between each of the 2 biomarkers and pathological subtypes, TNM stage, the presence of locoregional recurrence, and 5-year survival rate. RESULTS: p53 protein nuclear staining was positive in 49 patients (62%). The mean PCNA LI was 55.6%, ranging from 3.35% to 92.9%. High PCNA LI (>55.6%) might contribute to higher 5-year survival rate, but it did not reach statistical significance (P = .09). Positive p53 protein staining and low PCNA LI were associated with the presence of locoregional recurrence. No statistical significance was found between p53 protein expression and PCNA LI and pathological subtypes and TNM stage. CONCLUSION: p53 protein and PCNA LI were not an ideal prognostic indicator in predicting 5-year survival rate in nasopharyngeal carcinoma. Future work will direct toward searching for other potential biomarkers with the hope to reinforce prediction of prognosis.     

抑癌基因p16,p21及p53在鼻咽癌的表达

目的：观察抑癌基因ｐ１６,ｐ２１及ｐ５３在鼻咽癌（ＮＰＣ）的表达,探讨其与ＮＰＣ的发生发展及预后的关系。方法：用免疫组化ＳＰ法检测了１１６例ＮＰＣ组织和１５例无瘤鼻咽（ＮＰ）组织的ｐ１６,ｐ２１及ｐ５３基因的表达情况。     

p53家族新成员p63,p73与头颈部肿瘤

p53是研究较为广泛的抑癌基因，随着p53的广泛和深入研究，其家族成员p63与p73陆续被发现后，即成为人们关注的热点。本文就p63、p73的表达类型、结构和功能及其与头颈部肿瘤的相关性作一综述。     

Differential expression of p53, p63 and p73 proteins in human buccal squamous-cell carcinomas

Abnormalities in the p53 gene are regarded as the most consistent of the genetic abnormalities in oral squamous-cell carcinoma. Two new members of the p53 gene family, p73 and p63, have recently been identified, with the three sharing considerable sequence homology at the acidic N-terminal transactivation, central DNA-binding and C-terminal oligomerization domains, indicating possible functional and biological interactions. The differential expression of p73, p63 and p53 genes in human oral squamous-cell carcinoma does not yet appear to be completely understood, however. In this study, therefore, immunohistochemical analysis of protein expression was performed for 40 samples of well-differentiated human buccal squamous-cell carcinomas, with 10 specimens of normal buccal mucosa employed as controls. Differential expressions of p63, p73 and p53 proteins in the carcinoma samples were: p63+/p73+/p53 + (n = 28; 70%); p63+/p73+/p53- (n = 4; 10%); p63+/p73-/p53- (n = 8; 20%), respectively; and p63+/p73+/p53- for normal mucosa (n = 10; 100%). A significant correlation between p53, p63 and p73 immunoexpression was demonstrated for the buccal squamous-cell carcinoma samples (P < 0.0001; Fisher's exact test). Significance was not achieved for the correlation between p73 and p53 immunoexpression and clinicopathological parameters for buccal carcinomas (P > 0.05; Fisher's exact test). Our results indicate that both p73 and p63 may be involved in the development of human buccal squamous-cell carcinoma, perhaps in concert with p53.     

p53、p63、p73在鼻腔鼻窦恶性肿瘤中表达的临床意义

目的:探讨p53、p63、p73在鼻腔鼻窦恶性肿瘤中表达的临床意义及相关性。方法:用免疫组织化学方法检测67例鼻腔鼻窦恶性肿瘤、36例癌旁组织和36例鼻腔鼻窦的非癌组织中p53、p63和p73的表达。结果:p53、p63在鼻腔鼻窦恶性肿瘤中的阳性表达率明显高于癌旁和非癌组织(均P<0.01),p53与p63在鼻腔鼻窦恶性肿瘤中的表达存在正相关(P<0.01);p73在鼻腔鼻窦恶性肿瘤、癌旁和非癌组织中的表达差异无统计学意义(P>0.05)。结论:p53与p63在鼻腔鼻窦恶性肿瘤的发生发展过程中可能存在相关性,是鼻腔鼻窦恶性肿瘤发生过程中的重要因素之一,而p73在鼻腔鼻窦恶性肿瘤的发生过程中可能不发挥作用。     

Evaluation of p53, p63, p21, p27, ki-67 in paranasal sinus squamous cell carcinoma and inverted papilloma

Using a molecular genetic approach, we try to confirm the molecular alterations of inverted papilloma and clarify its status as a putative precursor lesion of sinonasal squamous cell carcinoma. To better understand its genetics, we investigated the immunohistochemical protein expression patterns of cell-cycle-regulators p53, p63, p21, p27 and proliferation marker Ki-67 in 22 inverted papilloma and 9 squamous cell carcinoma of the sinonasal tract. Significantly elevated levels of p53 and p63 in squamous cell carcinoma of sinonasal tract compared with inverted papilloma were revealed. Ki-67-stained neoplastic cell nuclei were found in a significantly higher percentage of squamous cell carcinoma of sinonasal tract than in inverted papilloma, whereas no variation of p21 and p27 expression was identified. This work first examined the immunohistochemical overexpression of p63 in sinonasal inverted papilloma and squamous cell carcinoma. In conclusion, this is a first study shedding light on the expression of p63 in tumors of paranasal sinuses.     

p38丝裂原激活蛋白激酶通路对变应性鼻炎中黏蛋白5AC的信号转导机制初探

目的:探讨组胺诱导的离体鼻黏膜组织中,p38丝裂原激活蛋白激酶(p38MAPK)信号通路及环氧合酶-2(COX-2)对黏蛋白5AC(MUC5AC)表达的影响,以期阐明变应性鼻炎(AR)中黏液过度分泌的病理学机制。方法:应用Western blot技术检测不同浓度梯度的组胺诱导及p38MAPK特异性抑制剂SB203580或COX-2特异性抑制剂NS-398干预前后,体外培养的鼻黏膜组织中p38MAPK、COX-2和MUC5AC的蛋白质表达变化规律。结果:p38MAPK、COX-2和MUC5AC在体外培养的健康鼻黏膜组织中呈微弱表达。组胺呈浓度依赖性诱导鼻黏膜组织中p38MAPK、COX-2和MUC5AC蛋白质表达增加;以不同浓度梯度的NS-398阻断COX-2后,观察到NS-398呈浓度依赖性减弱组胺对MUC5AC蛋白质表达的诱导,对p38MAPK表达无明显影响。以不同浓度梯度的SB203580阻断p38MAPK后,观察到SB203580呈浓度依赖性减弱组胺对COX-2和MUC5AC蛋白质表达的诱导。培养液中单独加入SB203580或NS-398对MUC5AC的蛋白质表达无明显影响。结论:组胺通过诱导p38MA...     

Differential expression of p53, p63 and p73 proteins in human buccal squamous‐cell carcinomas

Abnormalities in the p53 gene are regarded as the most consistent of the genetic abnormalities in oral squamous‐cell carcinoma. Two new members of the p53 gene family, p73 and p63, have recently been identified, with the three sharing considerable sequence homology at the acidic N‐terminal transactivation, central DNA‐binding and C‐terminal oligomerization domains, indicating possible functional and biological interactions. The differential expression of p73, p63 and p53 genes in human oral squamous‐cell carcinoma does not yet appear to be completely understood, however. In this study, therefore, immunohistochemical analysis of protein expression was performed for 40 samples of well‐differentiated human buccal squamous‐cell carcinomas, with 10 specimens of normal buccal mucosa employed as controls. Differential expressions of p63, p73 and p53 proteins in the carcinoma samples were: p63+/p73+/p53 + (n = 28; 70%); p63+/p73+/p53– (n = 4; 10%); p63+/p73–/p53– (n = 8; 20%), respectively; and p63+/p73+/p53– for normal mucosa (n = 10; 100%). A significant correlation between p53, p63 and p73 immunoexpression was demonstrated for the buccal squamous‐cell carcinoma samples (P < 0.0001; Fisher's exact test). Significance was not achieved for the correlation between p73 and p53 immunoexpression and clinicopathological parameters for buccal carcinomas (P > 0.05; Fisher's exact test). Our results indicate that both p73 and p63 may be involved in the development of human buccal squamous‐cell carcinoma, perhaps in concert with p53.     

喉癌细胞来源外泌体通过微小核糖核酸-21-5p促进巨噬细胞M2极化的作用及机制研究#br#

目的　研究喉癌患者术前及术后血清来源外泌体及其微小核糖核酸-21-5p（miR-21-5p）的变化，探讨喉癌细胞通过miR-21-5p促进巨噬细胞M2极化的机制。方法　收集正常人群血清以及喉癌患者术前和术后1个月及6个月血清样本，分离外泌体，纳米流式细胞仪揭示粒子粒径和数量，BCA和Western blot分析外泌体的含量和标记分子表达。建立人喉癌细胞系AMC-HN-8和巨噬细胞共培养体系 以及外泌体处理巨噬细胞体系，流式分析巨噬细胞极化情况，qRT-PCR检测外泌体和细胞中miR-21-5p水平。结果  喉癌术前和术后1个月及术后6个月血清来源外泌体CD9、CD63和CD81阳性表达，术前喉癌血清来源外泌体数量和蛋白浓度均高于对照组，而患者术后血清来源外泌体数量和蛋白浓度均比术前低。细胞水平发现AMC-HN-8与巨噬细胞共培养或者其来源外泌体处理巨噬细胞均降低了M1型巨噬细胞比例，提升M2型细胞比例。分子水平检测发现术前喉癌血清来源外泌体中miR-21-5p水平升高，术后则明显下降，AMC-HN-8与巨噬细胞共培养或其来源外泌体处理巨噬细胞均能升高巨噬细胞中miR-21-5p水平。抑制AMC-HN-8中miR-21-5p水平，其分泌的外泌体中miR-21-5p也受到抑制，处理巨噬细胞后，巨噬细胞中miR-21-5p水平和M2型细胞比例下降，而M1型细胞比例有所恢复。结论  喉癌患者血清外泌体含量以及miR-21-5p水平较高，喉癌细胞可通过外泌体传递miR-21-5p作用于巨噬细胞，促进巨噬 细胞M2极化。     

Expression of p63 and p73 in acoustic neuroma and its possible clinical relevance

ObjectivesAssess p63 and p73 expression in acoustic neuroma and its correlation with clinical and radiological findings.Materials and methodsmedical records of 34 patients who were operated on for acoustic neuroma during a 3-year period (2001-2003) were evaluated retrospectively. Immunohistochemical analysis of the schwannoma was performed with p63 and p73 antibodies and clinical patient characteristics were correlated with the immunoreactivity results.Results41% of the acoustic neuroma specimens showed p63 and p73 staining. Correlation between both proteins was 100%. Age of the patients tended to be older when staining was positive, but no statistical significance was achieved. Likewise, tumour size was bigger for positive tumours but, again, this difference was not statistically significant. There was no correlation between gender and immunostaining.Discussion and ConclusionsExpression of p63 and p73 was demonstrated in almost half of the patients studied. Although both proteins were more prevalent in older patients and bigger tumours, this difference was not statistically significant, probably due to the reduced sample size. No differences were found in laterality, gender or audiogram. However, the expression of these two proteins in almost half of the tumours shows that they can play a role in the development and progression of acoustic neuromas, although further studies are needed.     

p15和p27蛋白在鼻咽癌中的表达及其相关意义

目的研究周期素依赖激酶蛋白质类p15和p27蛋白在鼻咽癌（nasopharyngeal carcinoma,NPC）组织中的表达,探讨它们之间的关系与NPC生物学行为及预后的相关性。方法应用免疫组织化学EnVision两步法检测43例NPC组织和21例鼻咽黏膜慢性炎症组织,即非癌鼻咽组织（non-tumor nasopharyngeal,NP）中p15和p27表达水平。结果①NPC组织中p15和p27阳性表达率分别为65.1%和69.7%,与NP组比较,差异有统计学意义（P〈0.05）;②p15和p27蛋白在NPC中的表达与NPC临床分期、淋巴结转移、脑神经侵犯及治疗后5年生存率无关（P〉0.05）;③p15、p27阳性表达之间有相关性（P〈0.05）。结论p15蛋白和p27蛋白的表达缺失对NPC的发生起一定作用。     

抑癌基因p14ARF与p32在喉鳞状细胞癌中的表达及其意义

目的 观察p14ARF与p32在喉鳞状细胞癌（laryngeal squamous cell carcinoma,LSCC）组织中的表达情况及其与临床病理特征的关系,探讨其对LSCC发 生发展过程中的影响。方法 分别用荧光定量PCR和免疫组化法检测了50例LSCC组织及对应的癌旁组织中p14ARF与p32的mRNA和蛋白质的表达。采用SPSS 17.0软件进行统计分析。结果 p14ARF mRNA在LSCC组织中的表达水平与肿瘤临床分期相关（χ 2=20.704,P <0.05）,并呈现早期相对上升,晚期相对下降的趋势。p32 mRNA在LSCC组织与癌旁组织中的表达无明显的相关性（t =-0.684,P >0.05）p14ARF蛋白定位肿瘤细胞的细胞质和细胞核,p32蛋白主要定位于细胞浆,两者在蛋白水平上的表达结果与PCR基本一致。p14ARF与p32在LSCC组织中的表达无关联性（r =0.157,P >0.05）。结论 p14ARF在LSCC中的异常表达提示其可能参与了肿瘤的早期发生发展过程,并有可能成为LSCC早期诊断的一个分子标志物,而p32可能在LSCC的发生发展过程中并未起到关键的作用。