鼻咽癌患者鼻咽组织和血清中EBV检测

目的：比较血清爱泼斯坦－巴尔病毒（ＥＢＶ）抗体滴定度测定与癌组织ＥＢＶ基因组检测对鼻咽癌（ＮＰＣ）的诊断价值。方法：１４６例患者采用双盲法测定血清ＥＢＶ－ＶＣＡ－ＩｇＡ,ＥＢＶ－ＥＡ－ＩｇＡ和活检组织ＥＢＶ－ＤＮＡ（ＰＣＲ）。全组病例按活检病理检查结果分析：ＮＰＣ组,非ＮＰＣ组（对照组）。结果：ＮＰＣ组中ＥＢＶ－ＤＮＡ（ＰＣＲ）,ＥＢＶ－ＶＣＡ－ＩｇＡ和ＥＢＶ－ＥＡ－ＩｇＡ的阳性率分别为９０．８％     

MDM 2基因在鼻咽癌中的表达及其与p53蛋白表达、EB病毒潜伏感染的关系

目的：探讨ＭＤＭ２基因在鼻咽癌（ＮＰＣ）中的表达情况及其与ｐ５３蛋白表达、ＥＢ病毒（ＥＢＶ）潜伏感染的关系。方法：运用非同位素原位杂交、免疫组化和多聚酶链反应技术对４６例ＮＰＣ、１２例慢性鼻咽炎症粘膜（ＣＩＮＥ）分别进行ＭＤＭ２ｍＲＮＡ、ＭＤＭ２蛋白、ｐ５３蛋白及ＥＢＶ－ＤＮＡ的检测。结果：４６例ＮＰＣ中,１４例ｍＲＮＡ及ＭＤＭ２蛋白高表达,３８例为ｐ５３蛋白阳性,４３例为ＥＢＶ－ＤＮＡ阳性,１２     

聚合酶链反应检测EB病毒亚型对区分鼻咽良恶性病变的价值

以ＥＢ病毒核抗原（ＥＢＮＡ）Ａ亚型特异性引物作聚合酶链反应（简称ＰＣＲ－Ａ）同时用Ａ、Ｂ亚型ＤＮＡ片段和Ｂａｍ－Ｗ片段为探针，与鼻咽活检组织ＤＮＡ作打点核酸杂交（简称ＤＢＨ－Ａ、ＤＢＨ－Ｂ和ＤＢＨ－Ｗ）。ＤＢＨ－Ｂ结果全部阴性，ＰＣＲ－Ａ“ＤＢＨ－Ａ和ＤＢＨ０Ｗ检测的阳性率，３１例鼻咽癌分别为８３．９％（２６／３１）、７４．２％（２３／３１）和８０．６％（２５／３１）；３３例非鼻咽癌分别为１５．２     

鼻咽癌活检组织及外周血白细胞EB病毒DNA的检测及其意义

目的：探讨鼻咽组织、外周血血白细胞ＥＢＶ基因组分布及与ＮＰＣ的关系。方法：采用ＰＣＲ法检测２６例ＮＰＣ,４例癌旁及１４例慢性鼻咽炎鼻 组织吸外周血白细胞中ＥＢＶ ＤＮＡ。同时采用酶免疫法测定血清ＶＣＡ－ＬｇＡ。结果：１．ＮＰＣ、癌旁及鼻咽炎组织ＥＢＶ ＤＮＡ无明显差异。２．ＮＰＣ与鼻咽炎组外周血白细胞ＥＢＶ ＤＮＡ有明显差异,Ｐ〈０．０５。３．ＮＰＣ与鼻咽炎组血清ＶＣＡ－ＩｇＡ之间存在明显差异。Ｐ     

γ—干扰素对喉癌细胞株HEP—2增殖活性的影响

用单克隆抗体Ｋｉ－６７（抗ＰＣＮＡ），以链霉菌素－生物素技术（ＬＳＡＢ）检测喉癌细胞株ＨＥＰ－２增殖细胞核抗原（ＰＣＮＡ）表达。人重组γ－干扰素（ｒｈｕ－ＩＦＮ－γ）抑制ＰＣＮＡ表达（即抗增殖活性）强弱与其剂量有关；雌激素对ｒｈｕ－ＩＦＮ－γ抗增殖作用无影响。提示：ｒｈｕ－ＩＦＮ－γ对喉癌细胞株ＨＥＰ－２有抗增殖作用，因而在喉癌的治疗中具有应用价值的。     

聚合酶链反应检测EB病毒亚型对区分鼻咽良恶性病变的价值

以ＥＢ病毒核抗原（ＥＢＮＡ）Ａ亚型特异性引物作聚合酶链反应（简称ＰＣＲ－Ａ），同时用Ａ、Ｂ亚型ＤＮＡ片段和Ｂａｍ－Ｗ片段为探针，与鼻咽活检组织ＤＮＡ作打点核酸杂交（简称ＤＢＨ－Ａ、ＤＢＨ－Ｂ和ＤＢＨ－Ｗ）。ＤＢＨ－Ｂ结果全部阴性，ＰＣＲ－Ａ、ＤＢＨ－Ａ和ＤＢＨ－Ｗ检测的阳性率，３１例鼻咽癌分别为８３．９％（２６／３１）、７４．２％（２３／３１）和８０．６％（２５／３１）；３３例非鼻咽癌分别为１５．２％（５／３３）、６９．７％（２３／３３）和４２．２％（１４／３３），其中２５例慢性鼻咽炎分别为１２．０％（３／２５）、９２．０％（２３／２５）和４０．０％（１０／２５）。鼻咽癌组与非癌组和慢性鼻咽炎之间，ＰＣＲ－Ａ阳性率的差异有非常显著的统计学意义（Ｐ＜０．０００１），可作为区别鼻咽良恶性病变的一个有价值的参考指标。这为鼻咽癌的病毒发病学提供了有价值的线索，也使将来用ＰＣＲ早期诊断鼻咽癌成为可能。     

聚合酶链反应对颈部肿块的EBV检测

目的：探讨聚合酶链反应（ＰＣＲ）对隐匿性鼻咽癌的诊断意义。方法：采用ＰＣＲ检测５８例颈部肿块的细针抽吸标本中的ＥＢ病毒（ＥＢＶ）。结果：３５例颈中、上淋巴结转移癌２８例ＥＢＶ阳性,３例淋巴瘤阴性,４例锁骨上淋巴结转移癌阴性;１６例淋巴结炎性病变１例为弱阳性,１５例均为阴性;该法诊断鼻咽癌的灵敏度为８９．３％,特异性为８６．７％。结论：用ＰＣＲ检测颈部转移癌中的ＥＢＶ－ＤＮＡ,对隐匿性鼻咽癌的诊断具     

鼻、鼻窦恶性肿瘤中EB病毒和人乳头状瘤病毒的检测

目的：探讨ＥＢ病毒和人乳头状瘤病毒（ＨＰＶ）与鼻、鼻窦恶性肿瘤的关系。方法：用聚合酶链反应（ＰＣＲ）方法检测，３２例鼻、鼻窦恶性肿瘤组织蜡块的ＥＢ病毒和ＨＰＶ（ＨＰＶ６、１１、１６、１８、３３型）基因，分析其与病理分型及ＴＮＭ分期的关系。结果：３２例中检出ＥＢ病毒１２例（３７．５％），ＨＰＶ２１例（６５．６％），其中混合感染６例，５例均未检出。与之对照的１０例鼻息肉中未检出ＥＢＶ和ＨＰＶ。ＴＮＭ分     

分泌性中耳炎中耳积液的Epstein-Barr病毒测定

目的：探讨在分泌性中耳炎（ＳＯＭ）中的发病过程中是否有Ｅｐｓｔｅｉｎ－Ｂａｒｒ病毒（ＥＢＶ）参与。方法：应用聚合酶链反应（ＰＣＲ）技术对３４例ＳＯＭ患者（ＳＯＭ组）的血清、口腔含濑液和中耳积液（ＭＥＥ）进行ＥＢＶ检测,并与２０例正常人进行比较。结果：ＳＯＭ组血清和口腔含漱液标本中ＥＢＶ的检出率明显高于对照组,ＳＯＭ组ＭＥＥ中的ＥＢＶ检出率高于其血液标本。结论：在ＳＯＭ的发病过程中有ＥＢＶ的参与。     

Bcl—2,Bax蛋白产物在鼻咽癌中的表达,分布及意义

目的：探讨Ｂｃｌ－２、Ｂａｘ蛋白产物在鼻咽癌组织中的表达情况。方法：应用免疫组化技术Ｓ－Ｐ法,对４２例ＮＰＣ和１０例正常鼻咽粘膜组织标本,进行了调亡相关基因Ｂｃｌ－２和Ｂａｘ蛋白产物检测。结果：（１）在正常鼻咽粘膜中,Ｂｃｌ－２蛋白主要分布在基底层细胞,而Ｂａｘ蛋白分布在基底层以上细胞;（２）在ＮＰＣ组织中,Ｂｃｌ－２和Ｂａｘ蛋白产物表达阳性率为８５．７％和８３％,且Ｂｃｌ－２蛋白表达与ＢＰＣ颈淋     

Monitoring tumor recurrence with nasopharyngeal swab and latent membrane protein-1 and epstein-barr nuclear antigen-1 gene detection in treated patients with nasopharyngeal carcinoma

OBJECTIVES: Epstein-Barr virus (EBV) is closely related to nasopharyngeal carcinoma (NPC). Detection of EBV genomic DNA in a nasopharyngeal swab specimen may indicate the presence of NPC, and the EBV genomic DNA is only detected in patients with NPC and not in other head and neck cancers. This study aims to prove that detection of EBV genomic DNA by means of the latent membrane protein (LMP)-1 gene and the Epstein-Barr nuclear antigen (EBNA)-1 gene in the nasopharynx in NPC patients after radiation therapy indicates local recurrence of NPC. STUDY DESIGN: Prospective. METHODS: Nasopharyngeal swab with polymerase chain reaction (PCR)-based LMP-1 and EBNA-1 gene detection was used to monitor local recurrence in 84 NPC patients who completed radiation therapy. RESULTS: Of the 12 patients demonstrating positive LMP-1 and EBNA-1 gene, 11 had local recurrence, and 10 of them had early rT1 mucosal recurrence. Subsequent salvage nasopharyngectomy controlled local disease in nine. Only one local recurrence in the skull base failed to show LMP-1 gene initially. Detection of LMP-1 gene and later verification with EBNA-1 gene from nasopharyngeal swabs in NPC patients after radiation therapy predicted local recurrence with a sensitivity of 91.7% and a specificity of 98.6%. CONCLUSIONS: Nasopharyngeal swab with LMP-1 and EBNA-1 gene detection is a useful and reliable method to monitor local recurrence in NPC patients. It helps to detect recurrence early and may improve local control and enhance survival.     

Latent membrane protein-1 oncogene deletions in nasopharyngeal carcinoma in Caucasian patients

OBJECTIVE: The latent membrane protein-1 (LMP-1) is an Epstein-Barr virus (EBV)-transforming protein expressed in nasopharyngeal carcinoma (NPC). A 30-bp deletion in the LMP-1 oncogene has been described in NPC patients from Asia. The purpose of this study was to evaluate the association between NPC and such an EBV deletion in Caucasian patients. MATERIAL AND METHODS: Twenty-seven patients with a diagnosis of NPC were selected. Most of the NPCs were classified as Stages III and IV using the International Union Against Cancer system. Formalin-fixed, paraffin-embedded NPC specimens were found for these cases. Hematoxylin-eosin slides were reviewed and survival analysis was done using the log-rank method. In situ hybridization for EBV-encoded non-polyadenylated RNAs and expression of LMP-1 by means of immunohistochemistry was also performed. Polymerase chain reaction for LMP-1 oncogene analysis was performed to detect the presence of a 30-bp deletion in NPC specimens and EBV-related controls RESULTS: The 30-bp deletion was identified in 67% of NPC cases and in 30% of controls, a statistically significant difference (p = 0.01, chi2 test). LMP-1 deletion was not statistically associated with a worse prognosis in NPC patients (5-year survival: 33% in wild-LMP-1 strains vs 24% in deleted-LMP-1 strains; p = 0.053, log-rank test). CONCLUSION: A 30-bp deletion in the LMP-1 oncogene is present in more than half of Caucasian NPC cases EBV carrying partial deletions in the LMP-1 oncogene may play a role in the pathogenesis of NPC in Caucasian patients.     

Latent Membrane Protein-1 Oncogene Deletions in Nasopharyngeal Carcinoma in Caucasian Patients

Objective --The latent membrane protein-1 (LMP-1) is an Epstein-Barr virus (EBV)-transforming protein expressed in nasopharyngeal carcinoma (NPC). A 30-bp deletion in the LMP-1 oncogene has been described in NPC patients from Asia. The purpose of this study was to evaluate the association between NPC and such an EBV deletion in Caucasian patients. Material and Methods --Twenty-seven patients with a diagnosis of NPC were selected. Most of the NPCs were classified as Stages III and IV using the International Union Against Cancer system. Formalin-fixed, paraffin-embedded NPC specimens were found for these cases. Hematoxylin-eosin slides were reviewed and survival analysis was done using the log-rank method. In situ hybridization for EBV-encoded non-polyadenylated RNAs and expression of LMP-1 by means of immunohistochemistry was also performed. Polymerase chain reaction for LMP-1 oncogene analysis was performed to detect the presence of a 30-bp deletion in NPC specimens and EBV-related controls. Results --The 30-bp deletion was identified in 67% of NPC cases and in 30% of controls, a statistically significant difference (p = 0.01, χ² test). LMP-1 deletion was not statistically associated with a worse prognosis in NPC patients (5-year survival: 33% in wild-LMP-1 strains vs 24% in deleted-LMP-1 strains; p = 0.053, log-rank test). Conclusion --A 30-bp deletion in the LMP-1 oncogene is present in more than half of Caucasian NPC cases EBV carrying partial deletions in the LMP-1 oncogene may play a role in the pathogenesis of NPC in Caucasian patients.     

Detection of Epstein-Barr Virus in Nasopharyngeal Carcinoma by In Situ Hybridization and Polymerase Chain Reaction

The association of Epstein-Barr virus (EBV) with nasopharyngeal carcinoma (NPC) has been shown by various methods. The purpose of this study is to identify the most useful method to detect EBV in NPC. Both polymerase chain reaction (PCR) for EBV-DNA and in situ hybridization for EBV-encoded small RNAs(EBERs) were examined in formalin-fixed, paraffin-embedded NPC specimens. In situ hybridization was performed in 56 cases, and PCR for EBV-DNA was performed in 42 cases. EBV-DNA was detected in 0 of 3 keratinizing squamous cell carcinomas(KSCC), 22 of 24 nonkeratinizing carcinomas (NKC), all 13 undifferentiated carcinomas (UNPC), and 0 of 2 adenocarcinomas (AC). EBERs were detected in 0 of 5 KSCC, 30 of 32 NKC, 16 of 17 UNPC, and 0 of 2 AC. Among them, EBERs was detected in 35 of 42 cases in which PCR was also performed, 0 of 3 KSCC, 22 of 24 NKC, all 13 UNPC, and 0 of 2 AC, respectively. Both results were consistent in 40 of 42 cases. We conclude that both PCR and in situ hybridization are useful to detect EBV in NPC. In situ hybridization has a particular advantage because it can demonstrate the localization of EBV in neoplastic cells. In addition, close association of NKC and UNPC but not KSCC and AC with EBV is suggested.     

咽拭子检测潜伏膜蛋白1在鼻咽癌诊断中的应用

目的：验证咽拭子法检测EB病毒（EBV）潜伏膜蛋白1（LMP1）在鼻咽癌诊断中应用的可行性;检测LMP1野生型及30bp缺失变异型在鼻咽癌中的分布情况。方法：用咽拭子法收集鼻咽癌组及对照组鼻咽部脱落细胞,微量提取DNA,经PCR扩增人β珠蛋白基因序列验证咽拭子法的可行性,用特异性引物扩增出特异的LMP1序列,验证其在鼻咽癌诊断中的意义。测序分析LMP1变异情况。结果：咽拭子合格率为96．4％,LMP1基因作为鼻咽癌的检测指标,灵敏度为91．7％,特异性为95．6％,其中变异型LMP1在鼻咽癌中的表达频率为80．6％,野生型为11．1％。结论：咽拭子法可作为一种基因检测手段,LMP1基因的检测可以协同EB病毒壳抗原（EBVCA）-IgA作为鼻咽癌诊断的辅助指标,在LMP1（＋）的鼻咽癌患者中LMP1—30bp缺失突变普遍存在。     

鼻咽癌患者唾液和血清及外周血白细胞中EBV-DNA的定量检测

目的对鼻咽癌患者唾液、血清及外周血白细胞中的EBV-DNA进行检测,分析唾液、血清、外周血白细胞中EB病毒的DNA含量与鼻咽癌的相关性。方法对40例初诊鼻咽癌患者和50例正常人的唾液、血清及外周血白细胞中的EBV-DNA进行荧光聚合酶链反应(FQ-PCR)检测。结果EBV-DNA在鼻咽癌患者血清中的检出率为67.5%,显著高于正常人血清中的EBV-DNA检出率2%(χ2=44.48,P<0.001);鼻咽癌患者的唾液、外周血白细胞EBV-DNA检出率与正常人相比,无显著性差异;鼻咽癌患者和正常人的唾液EBV-DNA拷贝数与外周血白细胞EBV-DNA拷贝数呈正相关性;鼻咽癌患者血清EBV-DNA拷贝数与TNM分级有关,分级越晚含量越高。结论 EB病毒长期存在于多数人的唾液中,也可存在于外周血B淋巴细胞中。血清EBV-DNA检测在鼻咽癌的临床诊断中具有较高的敏感性和特异性,有助于鼻咽癌的早期诊断;血清EBV-DNA拷贝数与TNM分级有关,可在分子水平对鼻咽癌的TNM分级进行补充。     

鼻咽癌组织中程序性死亡配体-1的表达及其临床意义

目的:探讨程序性死亡配体-1(PD-L1)在鼻咽癌(NPC)中的表达及其临床意义.方法:采用免疫组织化学染色法检测59例NPC患者的癌组织和10例正常鼻咽黏膜组织中PD-L1的表达,RT-PCR技术检测30例NPC患者的癌组织及其中10例距离肿瘤组织3 cm以外的正常鼻咽黏膜组织中PD-L1mRNA的表达,并对其表达水平与临床病理组织学参数之间的关系进行相关性分析.结果:正常鼻咽黏膜组织中不表达PD-L1分子,而NPC癌组织中免疫组织化学PD-L1阳性表达率为67.8%(40/59),RT-PCR检测PD-L1mRNA的阳性表达率为66.7%(20/30).PD-L1的表达与肿瘤分期及淋巴结转移明显相关(P<0.05),但与患者的年龄、性别无明显相关(P>0.05).结论:PD-L1 NPC组织中的高表达,可能在肿瘤的发生、发展中起到一定的作用,有希望成为NPC免疫治疗的一个新靶点.     

EB病毒DNA与鼻咽癌关系的动态研究

目的：探讨EB病毒DNA（EBV—DNA）在鼻咽癌放疗前后的动态变化及与复发、远处转移的关系。方法：采用PCR加限制性内切酶酶切技术检测EBVDNA。结果：放疗前,74例标本中有71例（95．9％）EBV—DNA片段检出阳性;放疗50Gy／5周时,23例鼻咽原发灶和颈部淋巴结消失,其阳性率为13．0％（3／23）,余51例肿块未消者阳性率为62,7％（32／51）;放疗至70Gy／7周时,7例放疗后有残留,有残留者在放疗结束时EBV—DNA片段的检出阳性率为71．4％（5／7）,在67例肿块消失者中未检出阳性EBV—DNA片段。12例复发者中,11例EBV-DNA片段检出阳性;8例转移者中EBV—DNA片段检出均为阳性。结论：检测血浆EBV—DNA能很好地反映肿瘤的消长,是诊断鼻咽癌残留、复发及远处转移的敏感指标。     

免疫球蛋白kappa轻链基因在鼻咽癌中表达的初步研究

目的：为了探讨Ｉｇｋ是否参与了鼻咽癌的癌变过程及可能机制，对Ｉｇｋ在鼻咽癌中的表达进行了研究，并初步探讨Ｉｇｋ与ＥＢ病毒潜伏膜蛋白基因ＬＭＰ－１在鼻咽癌中的相关性。方法：分别运用原位杂交技术和免疫组化技术检测鼻咽癌活检组织中Ｉｇｋ的ＲＮＡ和蛋白质水平的表达。Ｗｅｓｔｅｒｎ印迹分析两株鼻咽癌细胞系（ＣＮＥ１和ＣＮＥ＿ＣＭＰ）ｋａｐｐａ蛋白的表达及强度差异。结果：原位杂交技术显示１００％（４２例／４２