鼻咽癌患者血清EB病毒DNA片段的检出及临床意义

目的：探讨鼻咽癌患者血清Epstein-Barr病毒(EBV)DNA片段检出率及其和EBV衣壳抗原IgA类抗体(VCA／IgA)的关系。方法：聚合酶链反应(PCB)加限制性内切酶酶切技术检测疱疹病毒的DNA，酶免疫组化技术检测EBV VCA／IgA抗体。结果：33例鼻咽癌患者中，抗体阳性率88％，DNA阳性率55％；对照组120例，仅1例检出EBV DNA片段。鼻咽癌患者中EBV DNA片段的检出率显著性高于对照组，但EBVDNA片段的检出率低于EBV VCA／IgA抗体的检出率。结论：进一步证实EBV与鼻咽癌有密切关系；EBV DNA片段及EBV VCA／IgA抗体检测，有利于鼻咽癌的辅助诊断。     

Detection of serum Epstein-Barr virus antibody level by ELISA in normal populations and nasopharyngeal carcinoma patients in Zhongshan City of China]

OBJECTIVE: To understand the distribution of Epstein-Barr virus (EBV) antibodies, namely EBNA 1/IgA, ZEBRA/IgA, EBNA 1/IgG in the populations of different ages or genders in Zhongshan City of Guangdong Province, China. METHOD: A total of 484 serum samples were obtained from the population of Zhongshan City including 16 patients with nasopharyngeal carcinoma (NPC) and were assayed with enzyme-linked immunosorbent assay (ELISA) for EBV antibodies. RESULTS: The serum antibody level of EBNA1/IgA, represented by its relative optical density value abbreviated as D(lambda), was 0.726 +/- 0.541 in male and 0.563 +/- 0.340 in female subjects, showing significant difference (P<0.001). The level of ZEBRA/IgG in the male subjects was significantly lower than that of the female subjects (0.709 +/- 0.480 vs 0.829 +/- 0.480, P<0.01). EBNA1/IgG levels differed little between the male and the female subjects, being 0.781 +/- 0.285 and 0.784 +/- 0.302 respectively (P<0.05). The positivity rate of ZEBRA/IgG was obviously related to the subjects' ages with a correlation factor of 0.766 (P<0.05), and NPC patients had significantly higher levels of the 3 antibodies than did the healthy subjects (P<0.01). CONCLUSIONS: The serum levels of these 3 antibodies fluctuate within a limited range in different age groups or between genders, significant increasing in NPC group and therefore may help the diagnosis of NPC. It can be imperative to set differently critical values of EBV antibody for different population groups to facilitate nasopharyngeal carcinoma (NPC) diagnosis and high-risk screening.     

鼻咽癌及鼻咽部粘膜中EB病毒DNA的检测

目的 检测鼻咽部粘膜及鼻咽中的ＥＢＶ ＤＮＡ片段,作为癌前病变的确立和癌变风险的评估指标。方法 用ＰＣＲ扩增ＥＢＶ的Ｂａｍ ＨＩＷ片段。结果 鼻咽癌中阳性率为８７．９％（１０２／１１６）,颈淋巴结转移灶６６．６％（２／３）,鼻咽癌放疗后鼻咽生炎７５．６％（２８／３７）,鼻咽粘膜不典型增生７１．１％（１５／２１）,鼻咽粘膜慢性炎和正常鼻咽粘膜上皮分别为２１％及２０％。结论：ＥＢＶ感染在鼻咽癌发生过程     

血浆EB病毒DNA含量定量检测对鼻咽癌的诊断

目的分析血浆EB病毒DNA定量测定对鼻咽癌的诊断价值。方法收集2007年2月-2008年5月在本院确诊的初诊鼻咽癌患者45例以及健康对照者45例，分别采集血浆标本，应用Real—timePCR方法检测EB病毒DNA含量，比较健康者和初诊鼻咽癌患者血浆EB病毒DNA拷贝含量水平的差异。结果鼻咽癌患者血浆EBV—DNA阳性率和复制量均高于健康对照者。两组阳性率和中位浓度的比较均有统计学差异。结论血浆EB病毒DNA定量检测是一种敏感而可靠的实验方法，对鼻咽癌的诊断具有重要价值。     

鼻咽癌病人多项免疫功能及EB病毒PCR检测的综合研究

为进一步了解ＥＢ病毒与鼻咽癌的关系,全面综合调查鼻咽癌病人的免疫功能,应用ＰＣＲ方法检测鼻咽病人组织块ＥＢ病毒ＤＮＡ,并对处国咽癌病人血清中ＥＢ病毒（ＥＢＶ－ＶＣＡ／ＩｇＡ）抗体、超氧化物歧化酶（ＳＯＤ）活性、脂质过氧化物降解产物（ＭＤＡ）及数种微量元素含量进行测定。结果：从鼻咽癌病人鼻咽部活检组织中成功地检测出ＥＢ病毒ＤＮＡ片断。进一步证实了ＥＢ病毒与鼻咽癌的发生有密切关系。并对鼻咽癌病人血清Ｖ     

血浆EBV DNA检测对鼻咽癌的诊断作用

目的探讨血浆EBV DNA检测对鼻咽癌的诊断作用。方法将2006年12月至2007年3月在我科就诊、病理确诊的65例鼻咽癌作为治疗前组,经我院放疗科治疗后,分别于放疗结束后3、5、8个月来我科系统复查的病人作为治疗后组,将同期行健康体检的29例作为对照组,共3组。采用荧光定量PCR方法对3组的空腹血浆EBV DNA进行检测。结果3组间血浆EBV DNA阳性率比较有统计学意义(P<0.05);治疗后组的阳性病例全部有鼻咽癌复发,阴性病例没有复发;有、无淋巴结转移组间阳性率比较差异有统计学意义(P=0.00)。结论血浆EBV DNA是鼻咽癌诊断的重要分子标记,其阳性的病例有较高淋巴结侵袭发生率。     

Analysis of Epstein-Barr viral DNA load,EBV-LMP2 specific cytotoxic T-lymphocytes and levels of CD4^＋CD25^＋ T ceils in patients with nasopharyngeal carcinomas positive for IgA antibody to EBV viral capsid antigen

Background Epstein-Barr virus （EBV） is a herpesvirus commonly associated with several malignant diseases including nasopharyngeal carcinoma （NPC）, which is a common cancer in Southeastern Asia. Previous studies showed that plasma levels of EBV-DNA might be a sensitive and reliable biomarker for the diagnosis, staging and evaluating of therapy for NPC. There are a few analyses of the levels of EBV-latent membrane protein 2 （LMP2）-specific cytotoxic T-lymphocytes （CTLs） in patients with NPC. This study was conducted to investigate the levels of EBV-LMP2-specific CTLs, EBV-DNA load and the level of CD4^＋CD25^＋T cells in such patients.
Methods From February 2006 to April 2006, 62 patients with NPC, 40 healthy virus carriers positive for EBV viral capsid antigen （EBV-IgA-VCA） and 40 controls were enrolled in the study. We used a highly sensitive ELISPOT assay, real-time polymerase chain reaction （PCR） and flow cytometry to measure the EBV-LMP2-specific CTL response, the EBV DNA load and the level of CD4^＋CD25^＋T cells, respectively.
Results The EBV-LMP2-specific CTL responses of the samples from the control, healthy virus carriers and patients with NPC were significantly different from the LMP2 epitopes, with the control and healthy virus carrier samples displaying a stronger response in three cases. There were significant differences in EBV DNA load in serum between NPC and the healthy groups; patients with NPC at stages Ⅲ or Ⅳ had significantly higher viral loads compared with those at stages Ⅰ or Ⅱ. A significantly higher percentage of CD4^＋CD25^＋ T lymphocytes were detected in the patients, compared with healthy virus carriers and healthy controls. Moreover, patients with advanced stages of NPC （Ⅲ and Ⅳ） had significantly higher percentages than the patients with early stages （Ⅰ and Ⅱ）. Conclusions Patients with NPC are frequently unable to establish or maintain sufficient immunosurveillance to control proliferating B cells harboring EBV and to destroy the tumor cells that express immunodominant LMP2 proteins.
Controlling the activity of CD4^＋CD25^＋T cells and elevating CD8^＋ cells specific for LMP2 epitopes could be an effective immunotherapy for patients with NPC.     

Serological diagnosis of nasopharyngeal carcinoma by enzyme linked immunosorbent assay: optimization, standardization and diagnostic criteria

OBJECTIVE To produce an enzyme linked immunosorbent assay (ELISA) to detect antibodies against Epstein Barr virus (EBV) specified nuclear antigen 1 (EBNA 1) and the 18 kD EBV matrix protein, and to determine and optimize its sensitivity and specificity for the diagnosis of nasopharyngeal carcinoma (NPC).

METHODS We used a combination of highly purified glutathione transferase fusion proteins of the 40 kD carboxy domain of EBNA1 and the 18 kD EBV matrix protein for coating ELISA plates. In three separate studies, we tested for IgA antibodies in serum specimens from 28 EBV seronegative donors, 284 EBV seropositive donors and 160 newly diagnosed NPC patients. By comparing the sensitivity and specificity of diagnosis obtained for different cutoff values, we derived several quantitative parameters to evaluate assay performance, establish objective diagnostic criteria which optimize the intrinsic diagnostic capability of the assay and assess the significance of individual test results, respectively. Optimum cutoff optical density (OD) is defined as the cutoff OD where sensitivity of the assay equals its specificity, and resolution of the assay is indicated by the value of sensitivity (or specificity) determined at the optimum cutoff OD. Diagnosis of NPC was achieved by setting a cutoff zone at +/- 20% of this value.

RESULTS All the EBV seronegative donors tested were not reactive, and most of the EBV seropositive donors were weakly reactive, while the majority of NPC patients were moderately or strongly reactive. While the assay was thus shown to be specific for EBV, there was an overlap in the level of these serum antibodies between few individuals of the two latter groups. It was shown that the assay performed equally well in two separate studies conducted under different testing conditions and using different collections of sera in that assay resolution determined on these occasions were 86% and 87% respectively. Diagnosis of NPC can be achieved at the same expected sensitivity of 89% and 83% determined at the lower and upper limits of the cutoff zones, with the corresponding values of specificity being 78% and 91%. It was further shown in the third study that resolution of the assay can be increased to 90% using an assay produced with a higher concentration of the same antigens, and that diagnosis of NPC can be achieved at a higher sensitivity ranging between 86% and 95% at a corresponding specificity of 93% and 86%.

CONCLUSIONS After optimization and standardization, the ELISA can achieve a sensitivity ranging from 86% to 95%, with corresponding specificities of 93% and 86% respectively for the diagnosis of NPC.

     

Epstein-Barr virus infection in nasopharyngeal lymphoid hyperplasia

Ｏｂｊｅｃｔｉｖｅ　ＴｏｄｅｔｅｃｔｗｈｅｔｈｅｒＥｐｓｔｅｉｎＢａｒｒｖｉｒｕｓ（ＥＢＶ）ｈａｒｂｏｒｓｉｎｎａｓｏｐｈａｒｙｎｇｅａｌｌｙｍｐｈｏｉｄｈｙｐｅｒｐｌａｓｉａ（ＮＰＬＨ）ｗｈｉｃｈｉｓｆｒｅｑｕｅｎｔｌｙｔｏｂｅｓｅｅｎｉｎＧｕａｎｇｚｈｏｕ,ａｈｉｇｈｉｎｃｉｄｅｎｃｅａｒｅａｏｆｎａｓｏｐｈａｒｙｎｇｅａｌｃａｒｃｉｎｏｍａ（ＮＰＣ）,ａｎｄｔｏｅｘｐｌｏｒｅｔｈｅｒｅｌａｔｉｏｎｂｅｔｗｅｅｎＮＰＬＨａｎｄｄｅｖｅｌｏｐｍｅｎｔｏｆＮＰＣＭｅｔｈｏｄｓ　Ｔｗｅｎｔｙ…     

鼻咽癌外周血单个核细胞和血浆中EB病毒DNA水平变化及与放化疗疗效的关系

目的：比较鼻咽癌患者放化疗治疗前后外周血单个核和血浆EBV DNA水平的动态变化,探讨EBV DNA水平与鼻咽癌放化疗近期疗效的关系。方法采用实时荧光定量PCR方法分别测定整群选取的2015年7—11月福建省肿瘤医院放疗科收治的75例初诊鼻咽癌患者放化疗前后外周血单个核细胞和血浆EBV DNA水平,根据疗效评价分为CR、PR、SD、PD组,各组治疗前后外周血单个核细胞和血浆EBV DNA水平分别进行比较;对配对的75例治疗前鼻咽癌患者外周血单个核细胞和血浆EBV DNA水平进行检出率和相关性分析。结果 CR、PR组外周血单个核细胞和血浆EBV DNA治疗前后下降明显,差异有统计学意义(P<0.05);SD、PD组外周血单个核细胞和血浆EBV DNA治疗前后差异无统计学意义(P>0.05)。外周血单个核细胞和血浆EBV DNA总体检出率为78.7%、53.3%,差异有统计学意义(P<0.05);两种类型标本EBV DNA结果呈正相关(r=0.481,P=0.00)。结论外周血单个核细胞和血浆EBV DNA检测可作为评价鼻咽癌放化疗近期疗效的指标。     

咽后淋巴结转移鼻咽癌患者血浆EBV DNA和血清EB抗体与预后的关系

目的 探讨咽后淋巴结转移鼻咽癌（NPC）患者血浆Epstein-Barr病毒（EBV）DNA和血清EB病毒衣壳抗原IgA抗体（VCA-IgA）、EB病毒早期抗原IgA抗体（VEA-IgA）与疗效、预后的关系。方法 回顾分析139例咽后淋巴结转移NPC患者的临床资料,用qRT-PCR法检测血浆EBV-DNA水平,酶联免疫吸附试验检测EB病毒VCA-IgA、VEA-IgA水平,根据疗效评价分为缓解组和进展组,比较治疗前后三项指标水平差异,分析治疗前各指标与临床预后、总生存的关系,Kaplan-Meier法计算生存率,Cox回归模型进行预后因素分析。结果 不同T分期、N分期和临床分期的EBV DNA浓度,差异有统计学意义（P <0.05）。不同临床分期VCA-IgA浓度和VEA-IgA 浓度比较,差异有统计学意义（P <0.05）,不同年龄、性别的EBV DNA浓度、VCA-IgA浓度和VEA-IgA浓度比较,差异无统计学意义（P >0.05）。咽后淋巴结转移NPC患者治疗前后血浆EBV DNA浓度比较,差异有统计学意义（P <0.05）,治疗前后VCA-IgA浓度和VEA-IgA浓度比较,差异无统计学意义（P <0.05）。缓解组与进展组治疗前后EBV DNA差值比较,差异有统计学意义（P <0.05）,VCA-IgA和VEA-IgA差值比较,差异无统计学意义（P >0.05）。患者4年总生存率为72.7%,T₁期、T₂期、T₃期、T₄期的4年总生存率分别为89.5%、80.8%、71.4%和59.6%,差异有统计学意义（P <0.05）,有无远处转移、是否疾病进展和是否复发患者中位生存期和4年生存率比较,差异有统计学意义（P <0.05）。多因素Cox回归模型分析结果表明：临床分期［R=2.162（95% CI：1.130,3.215）,P <0.05］、EBV DNA水平［R=2.324（95% CI：1.242,5.529）,P <0.05］、颈部淋巴结转移侧数［R=3.012（95% CI：0.653,5.564）,P <0.05］是影响NPC生存率的危险因素。结论 EBV DNA、VCA-IgA及VEA-IgA可为咽后淋巴结转移NPC患者的疗效监测、预后判断和个体化治疗提供帮助。     

病毒性肝炎患者EBV感染的临床意义

目的：探讨ＥＢＶ感染对病毒性肝炎的作用及临床意义。方法：用ＥＬＩＳＡ法和ＰＣＲ法分别检测５７０例病毒性肝炎患者血清中ＥＢＶ－ＩｇＭ、ＩｇＧ和ＥＢＶ－ＤＮＡ。结果：①血清中ＥＢＶ现行感染率（ＥＢＶＩｇＭ和ＰＣＲ阳性）在乙型肝炎１３．０７％（２６／１９９），混合型肝炎２２．９５％（１４／６１），高于甲型３．３７％（３／８９），丙型９．９３％（１４／１４１）和戊型肝炎２．５０％（２／８０）（Ｐ＜０．０５，Ｐ＜０．０１）。②在慢性活动性肝炎１６．０６％（３５／２１８），重症肝炎１４．２８％（２／１４），高于急性肝炎４．７６％（８／１６８），慢性迁延性肝炎４．５０％（５／１１１）（Ｐ＜０．０５，Ｐ＜０．０１）。③４例ＥＢＶ感染的病毒性肝炎患者经干扰素治疗后，１例ＨＢＶ－ＤＮＡ转阴（１／４），而无ＥＢＶ感染患者３例治疗后２例转阴。结论：ＥＢＶ感染是病毒性肝炎慢性化、重症化的重要因素，并可能影响干扰素疗效     

巢式聚合酶链反应扩增外周血吞噬细胞内外伤寒沙门菌Vi基因

目的：明确外周血吞噬细胞内外伤寒沙门菌Ｖｉ基因检测在伤寒诊断中的意义。方法：高速离心沉淀,低渗溶解红细胞法分离８５例临床拟诊伤寒患者外周血吞噬细胞（内）及吞噬细胞外（血清中）伤寒沙门菌,巢式聚合酶链反应扩增伤寒沙门菌Ｖｉ基因;并与单管套式聚合酶链反应扩增粒细胞内伤寒沙门菌Ｈ基因、巢式聚合酶链反应扩增单核细胞内伤寒沙门菌Ｖｉ基因、１次聚合酶链反应扩增血清中伤寒沙门菌Ｈ基因比较。结果：巢式聚合酶链反应     

Serological diagnosis of nasopharyngeal carcinoma by enzyme linked immunosorbant assay:optimization,standardization and diagnostic criteria

Ｓｅｒｏｌｏｇｉｃａｌｄｉａｇｎｏｓｉｓｏｆｎａｓｏｐｈａｒｙｎｇｅａｌｃａｒｃｉｎｏｍａｂｙｅｎｚｙｍｅｌｉｎｋｅｄｉｍｍｕｎｏｓｏｒｂａｎｔａｓａｙ：ｏｐｔｉｍｉｚａｔｉｏｎ，ｓｔａｎｄａｒｄｉｚａｔｉｏｎａｎｄｄｉａｇｎｏｓｔｉｃｃｒｉｔｅｒｉ...     

巢式PCR与免疫组化在检测肾癌组织支原体中的应用

目的:评价巢式PCR和免疫组化技术在检测肾癌组织支原体中的应用和价值。方法:采用支原体通用引物巢式PCR技术和单克隆抗体PD4免疫组化技术,检测95例肾癌组织中支原体DNA和支原体p40蛋白表达。结果:免疫组化检测和巢式PCR检测的阳性检出率分别为64.2%和81.1%,巢式PCR技术检测肾癌组织支原体在灵敏度、特异度、约登指数、阳性预测值和阴性预测值方面均优于免疫组化技术。结论:巢式PCR和免疫组化技术均可作为肾癌组织支原体的检测方法,巢式PCR较免疫组化检测敏感。     

鼻咽癌患者EB病毒检测的临床意义

目的探讨EB病毒（epstein-barr virus,EBV）抗体EB-VCA-IgA（EB病毒衣壳抗原抗体）,EB-EA-IgA（EB病毒早期抗原抗体）及血浆中EBV-DNA（EB病毒DNA）水平在监测鼻咽癌放疗后疗效、预后判断及复发转移中的临床意义。方法鼻咽癌患者76例,放疗前检测EB-VCA—IgA,EB-EA-IgA及EBV DNA。放疗后再次检测上述指标,并与临床及影像学检查结果比较。结果76例中,EB-VCA—IgA放疗前阳性71例,阴性5例;放疗后阳性69例,阴性7例。EB-EA-IgA放疗前阳性68例,阴性8例;放疗后阳性者64例,阴性12例。EBV-DNA放疗前检测阳性60例,阴性16例;放疗后阳性11例,阴性65例。其中,11例检出阳性者中2例证实为远处转移,8例为临床复发,故其肿瘤进展阳性预测值为90．9％。在65例阴性患者中,共有3例远处转移,8例复发,故阴性预测值为83．1％。64例患者血浆EBV-DNA水平与临床结果相符,故准确率为84．2％。结论血浆EBV-DNA水平的检测是监测放疗后鼻咽癌患者转移、复发的有效指标。而EB-VCA—IgA,EB-EA-IgA尚无法证明其是判断临床分期、疗效及复发的可靠指标。     

鼻咽癌放射治疗前测定血管内皮生长因子的意义

目的:于放疗前后检测鼻咽癌患者血清中血管内皮生长因子(vascular endothelial growth factor,VEGF)和β2微球蛋白的水平,探讨其变化及临床意义.方法:采集58例鼻咽癌患者和24名健康体检者血清,应用酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)测定VEGF水平,用时间分辨荧光免疫分析(time-resolved fluoroimmunoassay,TRFIA)方法测定β2微球蛋白水平,并对其变化及其与临床分期和患者生存期的关系进行分析.结果:鼻咽癌患者治疗前血清VEGF水平为(174.0±130.0)ng/L,稍高于健康体检者的(134.1±66.6)ng/L,但差异无统计学意义(P>0.05).鼻咽癌TNM分期T4和临床分期Ⅳa患者治疗前血清VEGF水平较高.治疗前血清VEGF水平>267.3 ng/L者,无远处转移生存期短(P<0.05).β2微球蛋白水平高的患者,中位生存期短,但差异无统计学意义.结论:鼻咽癌治疗前VEGF水平明显升高提示预后不良.     

EB病毒DNA在正常全鼻咽组织及鼻咽癌中的表现

作者应用原位核酸杂交技术，采用ＥＢ病毒ＢａｍＨＩ－Ｗ片段为探针，检测了湘南地区正常鼻咽粘膜、鼻咽低分化鳞癌和未分化癌，高分化和中分化鳞癌，颈淋巴结转移性低分化鳞癌，颈淋巴结其它部位转移癌及鼻咽临近区恶性肿瘤标本。ＥＢＶＤＮＡ阳性例数分别为０／１０个，２４／２６个，２／７个，６／６个，０／４个和４／２４个。同时，对上述病例进行血清ＥＢ病毒壳抗原免疫球蛋白Ａ抗体检测，低分化鳞癌和未分化癌阳性例数为２３／２６个，与ＥＢＶＤＮＡ检测结果一致，经统计学处理，两者具相关性。实验结果表明：（１）湘南地区鼻咽癌的发生中ＥＢＶ感染起非常重要作用，ＥＢ病毒与鼻咽低分化鳞癌及未分化癌关系密切。（２）血清ＥＢＶ壳抗原ＩｇＡ抗体检测可作为对鼻咽癌早期诊断的有效监测指标。（３）在颈部原发灶不明的转移癌中有ＥＢＶＤＮＡ片段的检出，对确定鼻咽癌的诊断具有一定的价值。     

ELISA法检测中国中山市正常人及鼻咽癌患者血清EB病毒抗体水平

目的观察EB病毒抗体 EBNA1/IgA、ZEBRA/IgG、EBNA1/IgG在不同年龄、不同性别的正常人群中的分布,并与鼻咽癌患者进行比较。方法用ELISA法在全自动酶联免疫分析系统上检测468例正常人和16例鼻咽癌患者的血清标本。结果正常人血清各抗体的水平[rD（λ）值]分别为:EBNA1/IgA男性0.726±0.541,女性0.563±0.340,男性高于女性（P<0.001）;ZEBRA/IgG男性0.709±0.480,女性0.829±0.480,女性高于男性（P<0.01）;EBNA1/IgG男性0.781±0.285,女性0.784±0.302,男女之间无显著性差异（P>0.05）。ZEBR/IgG的阳性率随年龄增高而升高,相关系数为0.766（P<0.05）。鼻咽癌患者上述3种抗体的血清水平分别为3.265±1.76、1.408±0.672、1.249±0.317,均明显高于正常人（P<0.01）。结论正常人群中EBNA1/IgA、ZEBRA/IgG、EBNA1/IgG3种抗体的血清水平存在年龄和性别上的差异,但波动在一定范围内;在鼻咽癌的临床诊断和高危人群筛查中有必要根据具体的人群界定不同的阳性临界值;鼻咽癌患者此3种抗体的血清水平均明显升高,对鼻咽癌的诊断有一定意义。     

鼻咽癌中EB病毒启动子Qp的突变及其功能学研究

【目的】 探讨鼻咽癌细胞中启动EB病毒核抗原1(EBNA1蛋白)表达的EB病毒Q启动子(Qp)的变异特征,比较有第62 225位点(g→a)和第62 422位点(g→c)两个点突变的Qp与原型Qp(B95.8型)的功能学差异及其生物学意义&#65377;【方法】 采用PCR的方法扩增29例鼻咽癌组织石蜡标本和14例健康成人外周血标本(总共43例)中的Qp序列,PCR产物测序后分析其突变情况,统计学分析Qp中的点突变与鼻咽癌的相关性&#65377;把突变型和原型Qp分别克隆到荧光素酶报告基因载体上,检测相对光强度(RLU),比较突变型与原型Qp启动转录的活性&#65377;使用染色质免疫共沉淀(ChIP)实验来比较突变型和原型Qp与Sp1蛋白的亲和力&#65377; 【结果】 通过PCR扩增和测序实验,证实Qp的突变与鼻咽癌有密切的相关性(P = 0.0395,< 0.05)&#65377;突变型Qp启动转录的活性明显高于原型(RLU之比约为2.5:1,P < 0.05),并且突变型Qp与Sp1的亲和力较原型Qp增强约1.52倍&#65377;【结论】 在鼻咽癌组织中,突变型Qp可能通过增强与Sp1亲和力的机制,使其启动转录的活性明显增强&#65377;Qp特定位点的突变在EB病毒对鼻咽上皮细胞的感染和转化过程中可能起了重要的作用&#65377;