鼻咽癌患者血浆 EBV DNA 水平和 VCA-IgA 检测的临床意义

目的:探讨鼻咽癌患者血浆EBV DNA水平和VCA-IgA联合检测对鼻咽癌早期诊断的临床价值。方法:用荧光定量PCR方法检测血浆EBV DNA水平,常规免疫酶法检测VCA-IgA抗体滴度。结果:68例鼻咽癌患者中,EBV DNA阳性率95.59,中位拷贝数93×104copies/ml,VCA-IgA抗体阳性率92.65,抗体滴度≥1:20;其他头颈肿瘤36例中,EBV DVA阳性率5.56,中位拷贝数为0copies/ml,VCA-IgA抗体阳率36.11,阳性滴度均≤1:20;健康对照组EBV DNA阳性率为35.56,其滴度除3例≥1:40外,余均≤1:20。鼻咽癌患者中EBV DNA和VCA-IgA抗体阳性率显著高于对照组。结论:进一步证实EBV与鼻咽癌有密切关系;EBV DNA水平和VCA-IgA抗体滴度联合检测,有助于鼻咽癌的早期辅助诊断。     

鼻咽癌病人血清中抗EB病毒膜抗体的测定

作者应用一种简便快速的活细胞间接免疫荧光法检测了127例鼻咽癌、其它恶性肿瘤病人及正常人血清中EBV MA—IgG、EBV MA—IgA抗体,并与EBV VCA—IgA抗体作了对比。鼻咽癌放疗前病人的EBV MA—IgG、MA—IgA抗体的阳性率和几何平均滴度与EBVVCA—IgA相似,明显地高于其它肿瘤病人和正常人。其中EBA MA—IgA抗体较EBV MA—IgG更为特异。因此EBV MA—IgG、MA—IgA抗体的检测对鼻咽癌的临床诊断是有意义的。放疗后鼻咽癌病人的EBV MA—IgA抗体水平随病人存活时间的延长而逐渐下降。当肿瘤复发或转移时又上升至放疗前的水平,故EBV MA—IgA抗体的检测对鼻咽癌病人预后的观察可以提供另一个血清学指标。     

耳、鼻、咽、喉

面神经监测显微手术切除听神经瘤；NP9基因抑制鼻咽癌细胞成瘤的实验研究；鼻咽癌患者EBV感染与细胞免疫水平关系的研究；MRI对鼻咽癌′92福州分期的影响；鼻咽癌变不同时期的体外三维培养模型建立的实验研究；915例单纯根治性放疗鼻咽癌分期系统比较及建议（对′92福州分期的校验）；5-杂氮-2′-脱氧胞苷对人鼻咽癌裸鼠移植瘤的抑制作用。     

血浆EB病毒DNA检测在鼻咽癌中的应用进展

EB病毒(Epstein-Barr virus,EBV)与鼻咽癌的发生、发展密切相关,采用PCR方法对鼻咽癌患者不同阶段的血浆EB病毒DNA (EBV DNA)进行定量检测可有效评估患者对治疗的反应,预测复发、转移的风险,治疗前的EBV DNA基线浓度与肿瘤负荷密切相关,而治疗后的EBV DNA含量与肿瘤复发转移关系更密切.EBV DNA定量检测有望成为一种新的肿瘤预后指标.本文就鼻咽癌患者不同时期的EBV DNA水平在诊断、分期、疗效评估和预后预测中的应用进行综述.     

EBV及PD⁃L1在局部晚期鼻咽癌中的研究进展

鼻咽癌是一种起源于鼻咽黏膜且与EB病毒（epstein-barr virus，EBV）感染有关的上皮性恶性肿瘤。血浆EBV DNA拷贝数在鼻咽癌分期、治疗、预后及随访监测中均有重要价值。鼻咽癌肿瘤组织中CD4+T细胞及CD8+T细胞浸润较多，肿瘤细胞表面程序性死亡配体-1（PD-L1）表达高达89%~95%，提示鼻咽癌肿瘤微环境多表现为免疫抑制状态。近年来，EBV DNA拷贝数及PD-L1表达与鼻咽癌肿瘤免疫微环境的关系也逐渐受到关注，本文就EBV与鼻咽癌免疫微环境的关系，以及EBV DNA拷贝数及PD-L1表达在局晚期鼻咽癌诊疗中的作用进行综述，以期为晚期鼻咽癌的精准免疫治疗决策提供依据。     

鼻咽癌初诊患者EB病毒与2008分期相关性的临床研究

[目的]探讨EB病毒(EBV)与鼻咽癌(NPC)2008分期的相关性;比较EBV IgA抗体滴度与EBV DNA拷贝数在NPC诊断与疗效评价中的差异.[方法]将初诊为NPC的患者98例,在放化疗前予以TNM分期;在放化疗前、后分别监测血清EBV抗体滴度与全血EBV DNA拷贝数.在放化疗后2个月,复查鼻咽部磁共振及全身检查后评价疗效.[结果]治疗前,血清EBV IgA抗体滴度阳性患者28例,全血EBV DNA拷贝数阳性42例.EBV IgA抗体滴度及全血EBV DNA拷贝数在不同TNM分期上差异均无统计学意义(P=-0.189,P=-0.074).全血EBV DNA拷贝数在Ⅲ期与Ⅱ期、Ⅲ期与Ⅳa期间比较,差异均具有统计学意义(P=0.024,P=0.022).放疗后,全血EBV DNA拷贝数转阴率高;但在提示转移或复发方面,EBV IgA抗体与EBV DNA比较二者差异无统计学意义(P=-0.095).[结论]血清EBV IgA抗体与NPC 2008分期无相关性.全血EBV DNA拷贝数与NPC 2008分期具有一定相关性.全血EBV DNA拷贝数转阴率高,优于EBV IgA抗体,在一定程度上能有效评价治疗疗效及监测病情变化.     

广东四会鼻咽癌患者治疗前EB病毒VCA／IgA抗体水平与生存的关系

背景与目的：Epstein-Barr病毒VCA／IgA抗体（serum immunoglobulin Aagainst Epstein-Barr vires capsid antigen,EBV—VCA／IgA）是目前应用最广泛的鼻咽癌诊断指标之一,但它对鼻咽癌预后判断的意义尚不明确。本研究探讨VCA／IgA抗体水平与鼻咽癌患者长期生存的关系,为确立VCA／IgA能否作为鼻咽癌独立的预后指标提供依据。方法：根据广东省四会市肿瘤发病与死亡登记资料,选择1990至2003年在中山大学肿瘤防治中心治疗的全部317例四会籍初诊鼻咽癌患者,收集患者的临床与病理资料,分析患者治疗前血清VCA／IgA抗体不同水平与生存期的关系。结果：在临床分期中Ⅲ、Ⅳ期的抗体水平率较Ⅰ、Ⅱ期为高,P=0．01。抗体滴度越高的患者生存时间越短,低水平组（〈1：160）的患者（n=170）与高水平组（≥1：160）患者（n=147）的5年生存率分别为65．0％和43．0％,P=0．01。多因素分析显示患者的临床分期、性别、治疗年代和EBV-VCA／IgA水平是影响生存期的独立因素。结论：鼻咽癌患者治疗前VCA／IgA抗体水平可能是影响患者生存的独立预后指标。     

EBV及HBV感染与鼻咽癌远处转移相关性探讨

目的研究EBV、HBV感染水平与鼻咽癌远处转移的关系.方法对118例证实远转和85例放疗后6年以上无瘤生存鼻咽癌患者,进行EBV抗体及乙肝病毒两对半检测.结果远转鼻咽癌患者的EBV-VCA-IgA抗体阳性率高于无转移患者(93.91%85.72%),但统计学差异无显著性(x2=3.66,P＞0.05);远转组中VCA-IgA抗体≥140者(57.41%)非常明显高于无转移组(39.39%),差异有显著意义(P＜0.05);远转组的EA-IgA阳性率(32.69%)明显高于无转移组(17.02%),差异有显著性(P＜0.05);远转组中合并HBV感染占27.53%,明显高于无转移组的13.43%(P＜0.05),且远转组中大三阳、e抗原或抗体阳性、C抗体阳性所占比例也明显多于无转移组.结论远转的鼻咽癌患者EBV和HBV感染比无转移患者活跃,提示EBV、HBV感染与鼻咽癌远处转移可能有-定关系.肿瘤防治杂志,2001,8(特)176-177     

鼻咽癌与EB病毒DNA研究进展

鼻咽癌与EB病毒(EBV)密切相关,通过检测鼻咽癌血浆中EBV DNA的表达水平,能够早期检测病灶残留、复发、远处转移,准确评价放化疗敏感性及判断预后,并可作为鼻咽癌临床分期的补充,以及成为分子靶向治疗的候选分子及鼻咽癌最重要的分子标志物。     

治疗前血浆EBV DNA在调强放疗鼻咽癌患者预后中的意义

摘 要：［目的］ 探讨初治鼻咽癌患者治疗前血浆EBV DNA表达水平的预后价值。［方法］ 回顾分析667例初诊鼻咽癌接受根治性调强放疗患者的临床资料,分析治疗前血浆EBV DNA与临床分期、总生存的关系。以1500拷贝数/ml为临界值,将患者分为EBV DNA高表达者和低表达者。Kaplan-Meier法计算生存率并用Log-rank法检验,多因素分析采用Cox模型分析。［结果］ Ⅰ、Ⅱ、Ⅲ、Ⅳ期患者中EBV DNA>0拷贝数/ml的比率分别为18.8%、34.4%、50.5%、66.4%。患者治疗前血浆EBV DNA的表达量与临床分期呈正相关（P<0.001）。临床分期和EBV DNA是患者总生存的独立预后因素。Ⅲ期患者中,EBV高低者两组差别具有统计学意义（χ2=4.084,P=0.043）,EBV DNA还是Ⅲ期患者总生存的独立预后因素。Ⅰ~Ⅱ期及Ⅳ期患者中,EBV DNA高低两组总生存率差别无统计学意义,但多因素分析发现,EBV DNA的绝对拷贝数是总生存的独立预后因素（P<0.05）。［结论］ 治疗前血浆EBV DNA对于各期鼻咽癌患者均具有预后作用,对于Ⅲ期患者,以1500拷贝数/ml为临界值,可以较好地判断预后,但对于Ⅰ~Ⅱ期及Ⅳ期患者,还需要更多的研究以确定合适的临界值。     

早期发现和筛选鼻咽癌的EB病毒血清学检测

鼻咽癌是中国人,特别是中国南方人的一种常见癌症;世界上除其他少数族群具有中等程度的发病率以外,它是一种罕见的癌症。鼻咽癌多见于男性,男女发病率之比约为3∶1,且好发于中年人。现今已有令人信服的证据,支持EB病毒(国际癌症研究协会归属为第一类人体肿瘤病毒)是鼻咽癌的致病因子,肯定参与鼻咽癌的多阶段、多因素发生过程。本文简要地复阅了EB病毒在鼻咽癌发病机理中的作用,重点地介绍了EB病毒抗体和EB病毒DNA作为鼻咽癌标志的主要应用;根据由鼻咽癌导致的EB病毒抗体反应的现代知识,并考虑到目前可采用的检测技术,提出了促进有效地早期发现鼻咽癌的血清学筛选策略。     

The association between circulating tumor cells and Epstein-Barr virus activation in patients with nasopharyngeal carcinoma

Background: Circulating tumor cells (CTCs) and microemboli (CTM) are attracting increasing attention in medical biology and clinical practice. However, the clinical relevance of CTCs in nasopharyngeal carcinoma (NPC) has not yet been ascertained, and no study has focused on the influence of Epstein-Barr virus (EBV) status on CTCs in NPC patients. These issues were therefore examined. Methods: Peripheral blood samples were prospectively obtained from 33 NPC patients before treatment. CTCs and CTM were captured using the Isolation by Size of Epithelial Tumor (ISET) method. Immunohistochemistry on CK5/6 (cytokeratin5/6) and P63, as well as in situ hybridization of EBERs (EBV-encoded RNAs) were used to validate the harvested tumor cells. Results: Out of 33 NPC patients, CTCs were detected in 22 cases (66.7%), and CTM were observed in 2 cases (6.1%). CTCs were presented in all stages of NPC patients but had no association with the TNM stages (all P > 0.05). The presence of CTCs appeared to correlate with EBV activation status. Among the total NPC patients, the EBV VCA-IgA levels in CTC-positive cases were higher than that in CTC-negative cases (negative vs. positive: 3.88 vs. 4.86, P = 0.016). A similar result was observed in the patients without distant metastasis (negative vs. positive: 3.76 vs. 4.95, P = 0.009). When the number of CTCs was considered, CTC counts were found to correlate with EBV VCA-IgA (R = 0.382, P = 0.041) and EBV DNA load (R = 0.396, P = 0.033) for all NPC patients as well as NPC patients without distant metastases. Conclusions: These findings strongly suggested detectable CTCs/CTM in all stages of NPC patients and support a positive correlation between CTCs and EBV activation in NPC patients.     

EB virus-associated antibodies in Caucasian patients with carcinoma of the nasopharynx and in long-term survivors after treatment

Sera from 30 Swedish patients with nasopharyngeal carcinoma (NPC) collected before or shortly after commencement of therapy and from 24 long-term survivors (LTS) were tested for antibodies to Epstein-Barr virus (EBV) capsid antigens (VCA), to the O and R components of the EBV-induced early antigen (EA) complex, and to EBV-determined cell membrane antigens (MA). In the tumor-bearing patients, all serologic reactivities increased overall from stage I to later stages of the disease: that is, with an increase in total tumor burden. In the LTS, who had remained apparently tumor-free for 3 to 29 years, all reactivities were substantially lower than in the tumor-bearing patients. While the data agreed in principle with those reported previously for Chinese NPC patients, there were considerable differences with respect to the incidence and titres of antibodies to the D component which, as discussed, might reflect a reduced responsiveness of Caucasian NPC patients or a difference in the distribution of various types of the disease among ethnic groups.     

血浆EBV DNA监测鼻咽癌治疗疗效意义

目的 探讨血浆EBV DNA监测鼻咽癌治疗疗效的临床意义。方法 回顾分析2016-2017年间本院初诊的799例鼻咽癌根治性调强放疗患者的临床资料。分析疗前血浆EBV DNA与临床分期、肿瘤进展的相关性,比较放疗结束及随访中EBV DNA与肿瘤进展的关系。结果 疗前DNA表达水平与临床分期、肿瘤进展呈正相关(P<0.001)。放疗结束后6~8周,19例(2.3%)血浆EBV DNA持续阳性者预后最差,14例发生了肿瘤进展。9例放疗结束后6~8周转为EBV DNA阴性,3例肿瘤进展。而放疗结束EBV DNA阴性患者肿瘤进展率仅8.3%(64/772),3个组无肿瘤进展生存率不同(P<0.05)。随访中持续性血浆EBV DNA阳性,诊断肿瘤进展的敏感性、特异性、准确性分别为77.6%、100%、98.1%。结论 鼻咽癌患者疗前EBV DNA表达水平与肿瘤负荷和肿瘤进展相关,放疗结束6~8周EBV DNA持续阳性者预后极差,应给予合适的辅助治疗。随访中持续性血浆EBV DNA阳性诊断肿瘤进展的正确性高,是鼻咽癌根治性治疗后可靠的疗效监测指标。     

血浆EBV DNA监测鼻咽癌治疗疗效意义

目的 探讨血浆EBV DNA监测鼻咽癌治疗疗效的临床意义。方法 回顾分析2016-2017年间本院初诊的799例鼻咽癌根治性调强放疗患者的临床资料。分析疗前血浆EBV DNA与临床分期、肿瘤进展的相关性,比较放疗结束及随访中EBV DNA与肿瘤进展的关系。结果 疗前DNA表达水平与临床分期、肿瘤进展呈正相关(P<0.001)。放疗结束后6~8周,19例(2.3%)血浆EBV DNA持续阳性者预后最差,14例发生了肿瘤进展。9例放疗结束后6~8周转为EBV DNA阴性,3例肿瘤进展。而放疗结束EBV DNA阴性患者肿瘤进展率仅8.3%(64/772),3个组无肿瘤进展生存率不同(P<0.05)。随访中持续性血浆EBV DNA阳性,诊断肿瘤进展的敏感性、特异性、准确性分别为77.6%、100%、98.1%。结论 鼻咽癌患者疗前EBV DNA表达水平与肿瘤负荷和肿瘤进展相关,放疗结束6~8周EBV DNA持续阳性者预后极差,应给予合适的辅助治疗。随访中持续性血浆EBV DNA阳性诊断肿瘤进展的正确性高,是鼻咽癌根治性治疗后可靠的疗效监测指标。     

Epstein-Barr virus DNA in serum/plasma as a tumor marker for nasopharyngeal cancer.

Nasopharyngeal cancer (NPC) constitutes a type of carcinoma encountered frequently in Southern China, among Eskimos of the Arctic region, and to a lesser extent in Southeast Asia. Because EBV DNA present in plasma or serum of NPC patients has proven to represent a promising noninvasive tumor marker, the present study was designed to determine the incidence of serum/plasma EBV DNA by nested PCR during various disease management stages. By this method, we could detect EBV DNA in plasma/serum of 98 of 167 NPC patients prior to treatment, compared with 10 of 77 samples derived from healthy blood donors serving as controls, with a similar prevalence observed in plasma versus serum. Investigation of 13 patients subjected to radiotherapy revealed plasma EBV DNA to persist in the plasma of one case, whereas among the remaining patients, it had vanished during the early phase of treatment. Finally, with 52 samples derived from 37 NPC patients during follow-up, we established 100% specificity and 0% false-positive rate for plasma DNA detection by nested PCR. Moreover, we subjected 24 known EBV DNA-positive serum samples to DNase digestion prior to DNA extraction and amplification to differentiate between free and encapsulated viral DNA, which demonstrated complete absence of the human beta-globin genomic DNA in contrast to EBV DNA detectable in 14 samples. In conclusion, applying this noninvasive method, serum/plasma EBV DNA constitutes a reliable tumor marker prior to, during, and after treatment of NPC.     

EBV DNA定量分析在监测鼻咽癌转移和复发中的临床意义

背景与目的:EB病毒(Epstein-Barrvirus,EBV)感染与鼻咽癌关系密切,近年来,有报道鼻咽癌患者血浆/血清中可检测到游离EBVDNA,但血浆EBVDNA水平对判断放疗后鼻咽癌患者转移、复发的临床意义尚缺少大宗研究报道。本研究定量检测鼻咽癌放疗后随诊患者血浆EBVDNA含量,探讨其在监测鼻咽癌转移、复发中的临床意义。方法:选择在中山大学肿瘤防治中心门诊随诊的放疗后鼻咽癌患者90例,用荧光定量PCR方法检测血浆EBVDNA含量,比较转移、复发与持续缓解患者血浆EBVDNA拷贝数。结果:放疗后转移或复发患者血浆EBVDNA的检出率为96.7%(29/30),中位拷贝数为2650copies/ml(0～5900000copies/ml);而持续缓解组患者血浆EBVDNA检出率12%(7/60),中位拷贝数为0copy/ml(0～71000copies/ml),差异均有统计学意义(P<0.01)。3例临床持续缓解但有血浆EBVDNA升高患者,在随后的3～4个月随访中,证实有肿瘤转移或复发。结论:血浆EBVDNA李宇红,等.EBVDNA定量分析在646定量检测可能成为监测放疗后鼻咽癌患者肿瘤转移、复发的敏感肿瘤标记物。     

Elevated serum levels of transforming growth factor beta1 in Epstein-Barr virus-associated nasopharyngeal carcinoma patients.

Nasopharyngeal carcinomas (NPCs) of non-keratinizing type are strongly associated with Epstein-Barr virus (EBV). EBV and its gene products induce the synthesis and/or release of transforming growth factor beta1 (TGF-beta1) from human cells and platelets. TGF-beta1 is an immunosuppressive cytokine, and many tumors are known to secrete it, to counter the host immune response. To determine the potential role of this cytokine in the pathogenesis of NPC, 53 serum samples from patients with EBV-associated NPC and 20 from healthy donors were analyzed for total and active TGF-beta content using ELISA. Serum samples for TGF-beta content were also analyzed from NPC patients at different clinical stages of the tumors. Significantly higher (p < 0.01) levels of active and total TGF-beta were found in the sera of NPC patients than in control sera. The ratio of active:total TGF-beta was also significantly (p < 0.01) increased in the NPC sera. Levels of this cytokine were also significantly higher (p < 0.05) in the sera of patients with advanced stages of tumor compared to patients with earlier stages. Furthermore, higher levels were seen in patients with relapsing than with remitting tumors; even higher levels were observed in NPC patients who died of the tumor. Our data suggest a role of this cytokine in the pathogenesis of NPC; therefore, it may prove to be a valuable biomarker molecule for the diagnosis and prognosis of NPC. Int. J. Cancer (Pred. Oncol.) 84:396-399, 1999.     

Early detection of nasopharyngeal carcinoma by plasma Epstein‐Barr virus DNA analysis in a surveillance program

BACKGROUND:

Nasopharyngeal carcinoma (NPC) is prevalent in Southeast Asia. Over the last decade, plasma Epstein‐Barr virus (EBV) DNA has been developed as a tumor marker for NPC. In this study, the authors investigated whether plasma EBV DNA analysis is useful for NPC surveillance.

METHODS:

In total, 1318 volunteers ages 40 to 60 years were prospectively recruited. Plasma EBV DNA and serology for viral capsid antigen immunoglobulin A (IgA) were measured. Participants who had detectable plasma EBV DNA or positive IgA serology underwent nasal endoscopic examination and a follow‐up plasma EBV DNA analysis in approximately 2 weeks. All participants were followed for 2 years to record the development of NPC.

RESULTS:

Three individuals with NPC were identified at enrolment. All of them were positive for EBV DNA and remained positive in follow‐up analysis. Only 1 of those patients was positive for EBV serology. In 1 patient who had NPC with a small tumor confined to the mucosa, the tumor was not detectable on endoscopic examination. Because of a 2‐fold increase in plasma EBV DNA on the follow‐up analysis, that patient underwent magnetic resonance imaging, which revealed the tumor. Among the participants who did not have NPC but had initially positive plasma EBV DNA results, approximately 66% had negative EBV DNA results after a median of 2 weeks.

CONCLUSIONS:

Plasma EBV DNA analysis proved useful for detecting early NPC in individuals without a clinical suspicion of NPC. Repeating the test in those who had initially positive results differentiated those with NPC from those who had false‐positive results. Cancer 2013. © 2013 American Cancer Society.     

Epstein-Barr virus in the pathogenesis of NPC

Epstein-Barr virus (EBV) is consistently detected in nasopharyngeal carcinoma (NPC) from regions of high and low incidence. EBV DNA within the tumor is homogeneous with regard to the number of terminal repeats. The detection of a single form of viral DNA suggests that the tumors are clonal proliferations of a single cell that was initially infected with EBV. Specific EBV genes are consistently expressed within the NPC tumors and in early, dysplastic lesions. The viral proteins, latent membrane protein 1 and 2, have profound effects on cellular gene expression and cellular growth, resulting in the highly invasive, malignant growth of NPC tumors. In addition to potential genetic changes, the establishment of a latent, transforming infection in epithelial cells is likely to be a major contributing factor to the development of this tumor.