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％     

Epstein-Barr virus in head and neck cancer assessed by quantitative polymerase chain reaction

OBJECTIVES/HYPOTHESIS: Epstein-Barr virus (EBV) has classically been associated with nasopharyngeal carcinoma and Burkitt's lymphoma. Recently, multiple studies have been published linking EBV with oral squamous cell carcinoma and, to a lesser extent, hypopharyngeal and laryngeal tumors. Using a sensitive method of detection, the authors sought to analyze the presence and quantity of EBV DNA in a large cohort of head and neck cancers. STUDY DESIGN:: Retrospective cohort study. METHODS: Three hundred head and neck cancer samples exclusive of nasopharyngeal carcinoma were examined for the presence of EBV using quantitative polymerase chain reaction. Eighty-four tumor samples from the larynx, 30 from the hypopharynx, 73 from the oropharynx, and 113 from the oral cavity were analyzed for EBV quantity, which was expressed as the number of viral copies per cell genome. Representative samples, which contained the highest EBV DNA levels, were examined using in situ hybridization. Results were correlated with tumor grade and site and tobacco and alcohol exposure. RESULTS: Three of 300 (1%) tumor samples were overtly positive for EBV DNA (defined as >0.1 copies of viral DNA/cell genome). Five of 300 (2%) tumor samples showed low levels (defined as >0.01 and <0.1 copies of viral DNA/cell genome), and 68 of 300 tumor samples (23%) showed trace levels (defined as < 0.01 copies of viral DNA/cell genome) of EBV DNA. No correlation was found between EBV positivity and tobacco exposure, alcohol exposure, or tumor grade. CONCLUSION: In the overwhelming majority of head and neck cancers in this North American cohort, EBV did not appear to contribute to growth of a dominant clonal population with integrated EBV genome and was unlikely to be a genetic etiological agent in tumor development. The low quantities of EBV detected in a minority of head and neck cancers may be related to the presence of EBV genome in rare lymphoid or epithelial cells adjacent to the primary head and neck cancer.     

Epstein-Barr virus latent membrane protein-1 (LMP-1) expression in oral squamous cell carcinoma

OBJECTIVES: Epstein-Barr virus (EBV) is frequently associated with malignant cell transformation through the action of the oncoprotein latent membrane protein-1 (LMP-1). The present study aimed to determine the presence of EBV in oral squamous cell carcinomas (OSCCs) and the expression of LMP-1 in neoplastic cells of EBV-positive OSCCs. STUDY DESIGN/METHODS: In a retrospective study of 78 OSCCs, we investigated the presence of the DNA of EBV by polymerase chain reaction, the expression of the oncoprotein LMP-1 by immunohistochemistry, and the presence of EBV-encoded RNA (EBER) by in situ hybridization. RESULTS: EBV DNA was detected in 19.2% of the cases. Expression of LMP-1 in neoplastic cells was found in 85.7% of the EBV-positive OSCCs. EBV presence was significantly more frequent (P <.05) in OSCCs localized on the lateral tongue. EBV-positive OSCCs more frequently presented (P <.05) greater nuclear atypia. CONCLUSION: EBV can appear in latent form in OSCC and express its main oncoprotein, LMP-1.     

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     

Immunohistochemical absence of CD21 membrane receptor in nasopharyngeal carcinoma cells infected by Epstein-Barr virus in Spanish patients

OBJECTIVES/HYPOTHESIS: The aim of this study was to analyze the relevance of the CD21 membrane receptor in nasopharyngeal carcinoma (NPC). CD21 is implicated in the introduction of the Epstein-Barr virus (EBV) genome into epithelial cells and B lymphocytes. STUDY DESIGN: Immunohistochemical analysis of CD21 in NPC. METHODS: Paraffin-embedded samples of NPC of different histological types with demonstrated presence of EBV were analyzed for CD21 expression using immunohistochemistry. RESULTS: We detected EBV by non-isotopic in situ hybridization (NISH) and nested polymerase chain reaction (PCR) in 100% of samples, regardless of histological type, supporting the previous view that all the types of NPC are variants of an EBV-associated malignancy. CD21 was not detected in NPC, and this absence was a typical feature in our data group. CONCLUSIONS: The loss of the CD21 membrane receptor could be one of the immunophenotypical changes of the neoplastic cells that occur in the evolution of the NPC malignancy.     

CK13基因与EB病毒DNA在鼻咽癌中的表达及临床意义

目的 探讨细胞角蛋白基因13（CK13）和EB病毒DNA在人鼻咽癌（NPC）组织中的表达及意义。方法 应用Northern杂交对32例NPC和8例慢性鼻咽炎（CINE）组织进行CK13mRNA水平检测,同时应用PCR方法进行EB病毒DNA检测。结果 ＣＫ１３基因在鼻咽炎组织中高表达,在ＮＰＣ组织中的表达显著下调。下调的频率达６５.63%(21/32)。结论 ＮＰＣ组织的分化可能与ＣＫ１３基因的表达下调有关。     

Adenotonsillar hypertrophy and Epstein-Barr virus in pediatric organ transplant recipients

OBJECTIVES/HYPOTHESIS: Epstein-Barr virus-related (EBV-related) lymphoid hyperplasia of the tonsils and adenoids is a precursor to post-transplantation lymphoproliferative disorder (PTLD). The incidence of post-transplantation adenotonsillar hypertrophy, a potential early sign of PTLD or EBV-related lymphoid hyperplasia, is not known. We sought to identify potential risk factors for adenotonsillar hypertrophy manifested as EBV-related hyperplasia and early PTLD in the pediatric solid organ transplant population. STUDY DESIGN: Cross-sectional analysis. METHODS: We developed a 65-point questionnaire concerning obstructive sleep disorder and upper respiratory tract infections and an 8-point focused physical examination, to identify prevalence of and risk factors for adenotonsillar hypertrophy in the pediatric transplant population. We evaluated 120 pediatric solid organ transplant recipients by parental questionnaire and focused adenotonsillar physical examination. RESULTS: Of the 120 patients, 62 had undergone liver transplantation and 58 had undergone kidney transplantation. Overall, the mean questionnaire score was 8.36 (range, 0-40) and the mean physical examination score was 3.86 (range, 1-8). Patients whose EBV serological test result was negative at the time of transplant had higher scores for both the questionnaire (mean score, 10.24) and the physical examination (mean score, 4.56) than those whose EBV serological test result was positive at the time of transplantation (scores of 7.38 and 3.30 for questionnaire and physical examination, respectively). The difference in examination scores was statistically significant (P <.003). CONCLUSIONS: Epstein-Barr virus seronegativity at the time of organ transplantation is a known risk factor for PTLD, with associated risk of developing EBV-related lymphoid hyperplasia. Our results indicate a higher incidence of symptoms and signs consistent with adenotonsillar hypertrophy in the EBV seronegative population. Adenotonsillar hypertrophy may be a precursor to EBV-related lymphoid hyperplasia and PTLD and must be identified in this patient population.     

用聚合酶链反应检测鼻咽癌脱落细胞中EB病毒DNA

应用聚合酶连反应（ＰＣＲ）技术检测３５例鼻咽癌患者（２４例刚进行￣（６０）Ｃｏ放疗或未治疗，１１例放疗接近结束或刚结束）的鼻咽部脱落细胞中的ＥＢ病毒ＤＮＡ，１２例急、慢性扁桃体炎或咽炎患者为正常对照组，另有４例鼻咽癌的活检组织。结果：２４例鼻咽癌未治疗或刚治疗组中脱落细胞ＥＢ病毒ＤＮＡ阳性２０例（８３．３％），阴性４例（１６．７％）；１１例鼻咽癌放疗将近结束或刚结束组ＥＢ病毒阳性１例（９．１％），阴性１０例（９０．９％）；１２例对照组ＥＢ病毒ＤＮＡ阳性１例（８．３％），阴性１１例（９１．７％）。４例鼻咽部活检组织ＥＢ病毒ＤＮＡ均阳性。说明应用鼻咽部刮片脱落细胞进行ＰＣＲ反应检测ＥＢ病毒ＤＮＡ是一种方法简单、痛苦少，特异性强、灵敏度高的检测手段，可用于鼻咽癌的诊断、普查、流行病学调查等。     

The 30-bp deletion of Epstein-Barr virus latent membrane protein-1 gene has no effect in nasopharyngeal carcinoma

OBJECTIVE: The specific 30-bp deletion of the Epstein-Barr virus (EBV)-derived latent membrane protein-1 gene has been suggested to be associated with the pathogenesis of nasopharyngeal carcinoma (NPC) and a more aggressive phenotype of some EBV-associated malignancies. METHODS: The authors conducted a retrospective cohort study. Between 1995 and 2001, 542 patients who had received complete courses of radiotherapy followed by at least 3 years of follow up were enrolled. Patients were divided into two groups according to the presence or absence of the 30-bp deletion in their tumor samples. RESULTS: A total of 446 (82.3%) patients were found to feature the 30-bp deletion, whereas 88 (16.2%) did not. Interestingly, dual infection was found in eight (1.5%) patients. No statistical significance was found in age, gender, NPC-presenting stage, radiosensitivity, and pathologic classification between the two groups. The actuarial 5-year overall survival rate and the distant metastasis-free rate for the 30-bp deletion and nondeletion group were not statistically different (61.3% vs. 65.4% and 68.1% vs. 73.1%; P = .132 and .135, respectively). In multivariate analysis, older age, nasopharyngeal recurrence, advanced tumor stage, and the development of distant metastasis were shown to be poorer prognosticators for overall survival, whereas the presence of 30-bp deletion was not. For distant metastasis, only advanced tumor stage was shown to be a poor prognosticator, whereas other variables, including the presence of 30-bp deletion, had no statistical significance. CONCLUSIONS: In this retrospective cohort, we have demonstrated that this specific 30-bp sequence deletion might be only the predominant variant rather than an NPC phenotype-associated polymorphism. Additionally, dual infection is a rare but possible phenomenon in the endemic NPC tumors.     

bcl-2、P 5 3基因及Epstein-Barr病毒在鼻咽癌中的表达

目的探讨鼻咽癌组织中p53、bcl-2、EBV表达情况及与主要临床指标间关系.方法免疫组化检测p53、bcl-2蛋白,多聚酶链反应(PCR)检测EBV.结果慢性炎性组织中p53、bcl-2及EBV阳性率分别为7.7%、23.1%和7.7%,明显低于鼻咽癌组织中的阳性率(分别为75.9%、88.9%和87.0%)(P<0.01).三者在鼻咽癌组织中共同阳性率为72.2%,p53、bcl-2明显正相关(r＝0.78,P<0.01).鼻咽癌伴颈淋巴结转移组p53阳性率(90.0%)明显高于无颈淋巴结转移组(58.3%)(P<0.01),临床Ⅱ、Ⅲ和Ⅳ期p53阳性率(分别为92.9%、78.9%和100%)均明显高于临床I期(38.5%)(P<0.01),且表达强度有增加的趋势.结论p53、bcl-2及EBV可作为鼻咽癌早期诊断的辅助指标之一,p53阳性的鼻咽癌患者较易发生颈淋巴结转移,随着鼻咽癌病程的发展,p53表达率及表达强度均增加,三者在鼻咽癌发生发展中起着重要作用.     

EB病毒Rta蛋白抗体检测对鼻咽癌病理分级的临床价值

目的探讨EB病毒Rta蛋白抗体对鼻咽癌病理分级的临床价值。方法选择我院诊治的鼻咽癌患者共计68例,选取同一时期来我院体检健康人54例,测定血清Rta-IgG水平,血清Rta-IgG与鼻咽癌病理分级采用spearman相关分析。结果7观察组(I期、II期、III期、IV期)血清Rta-IgG水平均高于对照组,鼻咽癌患者中I期、II期、III期、IV期血清Rta-IgG水平依次升高,有差异(P<0.05)。血清Rta-IgG水平与鼻咽癌病理分级之间呈正相关(P<0.05)。结论EB病毒Rta蛋白抗体对鼻咽癌病理分级有预测价值。     

Detection of Epstein‐Barr Virus–Derived Latent Membrane Protein‐1 Gene in Various Head and Neck Cancers: Is It Specific for Nasopharyngeal Carcinoma?

OBJECTIVE: The object of the study was to determine the incidence of the presence of Epstein-Barr virus-derived latent membrane protein-1 (LMP-1) gene in various head and neck cancers by polymerase chain reaction method. STUDY DESIGN: Prospective study. METHODS: During a 5-year period, polymerase chain reaction was used to investigate the presence of LMP-1 gene in various head and neck cancers from different locations and histopathological types, noncancerous nasopharyngeal biopsy samples, and tonsillectomy specimens from patients with chronic hypertrophic tonsillitis. RESULTS: Of 202 patients enrolled in the study, 53 were diagnosed by pathological study with oropharyngeal carcinoma, 45 with nasopharyngeal carcinoma, 26 with oral cavity carcinoma, 26 with laryngohypopharyngeal carcinoma, 31 with nasopharyngeal lymphoid hyperplasia, and 21 with tonsils with lymphoid hyperplasia. After the application of polymerase chain reaction, the LMP-1 gene was not detected in any sample from oral cavity carcinoma, laryngohypopharyngeal carcinoma, or nasopharyngeal lymphoid hyperplasia or from tonsillectomy specimens but was detected in only one case of tonsillar carcinoma. On the contrary, the LMP-1 gene was detected in 43 (95.6%) of 45 samples from patients with nasopharyngeal carcinoma. The statistical analysis shows a significant association (P <.001) between the presence of LMP-1 gene and tumor localization in the nasopharynx. CONCLUSIONS: The study shows that the presence of LMP-1 gene detected by polymerase chain reaction in the tumor cell is only significantly associated with tumor located in the nasopharynx, implying that Epstein-Barr virus plays a trifling role in the tumorigenesis of carcinomas arising from other head and neck locations. The polymerase chain reaction method that was used is a potential tool for screening nasopharyngeal carcinoma.     

Absence of nuclear p16 from Epstein-Barr virus-associated undifferentiated nasopharyngeal carcinomas

Objective: Epstein‐Barr virus (EBV) is detected in the majority of undifferentiated nasopharyngeal carcinomas (UNPCs, World Health Organization type III). However, the exact mechanism involved in the carcinogenesis of EBV‐associated UNPCs remains to be elucidated. An important unresolved question is: how is the normal cell cycle deregulated during EBV‐associated UNPC development? The p16^CDKN2 gene encodes a nuclear protein, p16, which inhibits the D‐type cyclin/cyclin‐dependent kinase complexes that phosphorylate the retinoblastoma gene product (pRb), thus blocking G₁ cell cycle progression. The objective of this study was to determine whether p16 absence is involved in the development of EBV‐associated UNPCs. Methods: We performed immunohistochemistry to detect p16 and pRb and in situ hybridization to detect EBV‐encoded small RNA (EBER) in UNPCs from 28 patients. Results: No p16 was detected in 23 of 28 UNPCs (82.1%), whereas pRb was expressed in all those examined and EBER was detected in 22 of 28 (78.6%). The absence of p16 was associated with the presence of EBER in UNPCs (P < .0001): none of the 22 EBER+ UNPCs expressed p16, whereas 5 of 6 EBER? UNPCs did. Conclusion: Our data suggest that loss of p16‐related cell cycle regulation plays an important role in the development of EBV‐associated UNPCs.     

Epstein-Barr virus (EBV) latent membrane protein 1 induces interleukin-8 through the nuclear factor-kappa B signaling pathway in EBV-infected nasopharyngeal carcinoma cell line

BACKGROUND/OBJECTIVES: Nasopharyngeal carcinoma (NPC) is a highly invasive and metastatic malignant tumor and is associated with Epstein-Barr virus (EBV) infection that exhibits type II latency. Angiogenesis is essential for tumor growth, invasion, and metastasis. Our previous studies have indicated that interleukin (IL)-8 was over-expressed in many NPC tissues and was found to be significantly correlated with angiogenesis by immunohistochemistry. STUDY DESIGN: In vitro design. METHODS: The influence of the EBV genome for IL-8 gene expression was studied using the EBV-genome-positive and -negative epithelial/NPC hybrid cell line NPC-KT. The EBV-positive and -negative clones were selected by polymerase chain reaction and in situ hybridization. RESULTS: EBV-positive clones expressed abundant IL-8 mRNA compared with EBV-negative clones. This result indicated that over-expression of IL-8 depended on the presence of EBV genomes in NPC-KT cells. Two encoded genes, latent membrane protein (LMP)1 and EBV-encoded small RNAs (EBERs), expressed in NPC were transfected in EBV-negative NPC-KT cells. LMP1 transactivated the IL-8 promoter, whereas EBERs did not. Moreover, the nuclear factor (NF)-kappa B binding site in the IL-8 promoter was essential for the response to LMP1, and the activator protein (AP)-1 binding site played only a partial role. CONCLUSIONS: LMP1 induces IL-8 mainly through the activation of NF-kappa B and partly through AP-1 in NPC model cell lines, NPC-KT, and this suggests that LMP1 plays an important role in the angiogenesis of NPC.     

鼻咽组织及脱落细胞中EB病毒对鼻咽癌诊断价值的对照研究

目的：探讨一种快速准确普查及诊断鼻咽癌（NPC）的方法。方法：应用聚合酶链反应（PCR）技术对临床怀疑NPC的144例患者的鼻咽组织和鼻咽脱落细胞中EB病毒（EBV）DNA进行检测,并与病理诊断对照。结果：PCR技术检测鼻咽组织中EBV DNA诊断NPC的敏感性和特异性分别为92．16％和80．95％;检测鼻咽脱落细胞中EBV DNA诊断NPC的敏感性和特异性分别为86．27％和71．43％,其差异均无统计学意义（均P〉0．05）。结论：检测鼻咽脱落细胞中EBV快速、简单,患者痛苦小,敏感性和特异性均较高,可用于NPC的诊断;对NPC的普查及流行病学调查尤其方便、实用,值得推广。对凡是EBV检测阳性的患者均应提高警惕,追踪观察。