人咽喉部肿瘤组织HPV免疫组化及DNA斑点杂交的检测

用免疫组化及ＤＮＡ斑点杂交技术检测人咽喉部乳头状瘤及鳞状细胞癌组织中ＨＰＶ壳蛋白抗原及ＨＰＶ６、１１、１６、１８型ＤＮＡ《结果显示，５例正常粘膜、１５例声带息肉ＨＰＶ抗原及ＤＮＡ检测均阴性。１１例乳状状瘤ＨＰＶ怕与ＨＰＶＤＮＡ阳性率均为４５．５％。２２例鳞状细胞癌ＨＰＶ抗原阳性率为２２．７％，ＨＰＶＤＮＡ阳性率为２７．３％，乳头瘤ＨＰＶ检出率与组织学的检查的结果相符。结果提示咽喉部乳头瘤及鳞状细胞     

子宫颈癌患者循环HPV DNA检测及临床意义的研究进展

近来，癌症患者血浆／血清中肿瘤DNA的临床意义和生物学特忡引起了人们的广泛注意：同样，人们对宫颈癌患者血浆／血清中的人类乳头状瘤病毒（HPV）的DNA亦作了研究。研究采用不同的检测标本（血浆／血清）、不同的DNA抽提方式及不同的引物来检测循环游离HPV DNA，所得的阳性率从6．9％～50％不等。因为宫颈癌患者血浆／血清中的HPV DNA很可能来源于肿瘤本身，这给我们提供了一个新的宫颈癌治疗后的监测工具，特别是针对晚期的患者。可以预测，这一极具有应用前景的分子肿瘤标志物不久将会在临床上被广泛应用。     

肿瘤组织中人乳头瘤病毒HPV基因组相关序列的检测

本文报道应用核酸分子杂交方法,以[α-(32)P]-dTTP标记的HPV16型基因组为探针,共检测恶性肿瘤53例(标本包括宫颈癌22例,阴茎癌3例、膀胱癌6例、肾癌3例、肺癌7例、胃癌1例、结肠癌11例)、7例良性肿瘤、8例正常上皮组织中HPV基因组相关序列。检测结果表明,在恶性肿瘤中仅于宫颈癌组织检测出HPV16型基因组相关序列59.1％,P<0.01。而良性肿瘤和正常上皮组织中均未检测出HPV16型和其他型别基因组相关序列。此种阴性结果,可能与地理分布、病变组织的病理类型、检测使用的探针和方法、例数少等因素有关。为进一步探讨HPV感染与肿瘤的关系,尚需深入研究。     

PCR技术检测复发性呼吸道乳头状瘤组织中的HPVDNA

应用多重引物ＰＣＲ技术对３６例复发性呼吸道乳头状瘤病（ＲＲＰ）和９例声带息肉石蜡包埋组织中的ＨＰＶ６／１１、１６、１８ＤＮＡ进行了检测。结果发现：（１）ＲＲＰ组织中ＨＰＶ６／１１ＤＮＡ的阳性率为６９．４％（２５／３６），其中喉乳头状瘤，口咽部乳头状瘤和鼻前庭乳头状瘤组织中ＨＰＶ６／１１ＤＮＡ的阳性纺分别为７０．４％（１９／２７）、６６．７％（４／６）６６．７％（２／３）；所有ＲＲＰ标本中均示发现特     

HPV16光敏生物素探针的制备及斑点杂交应用

近年来的研究表明,人乳头瘤病毒(HPV)很可能是子宫颈癌主要的病毒病因。目前,在HPV的分子生物学研究中,许多实验室都采用放射性同位素和生物素两种探针标记法。为了找到一种简单、快速、安全的标记探针方法,我们试用了光敏生物素来标记HPV16DNA作探针,并用斑点杂交法初步检测了部分宫颈癌组织中HPV的感染情况。 材料和方法 一、材料 1.试剂:6—氨基乙酸光敏生物素、蛋白酶K,内切酶BamHI,均为国产;DNA检测药盒(Bresa     

人乳头瘤病毒DNA在人喉癌细胞中表达的研究

为研究喉癌与人乳头状瘤病毒(HPV)感染的关系,本研究探讨了HPV在喉癌中的致病作用和基因组型的分布与表达。应用共有引物和多重引物PCR的方法,对160例喉不同病变的新鲜组织标本,进行HPV6、11、16、18、31、33、35、42、58共9型HPVsDNAs感染的检测。结果在喉癌组HPV感染的阳性率为49.3％(35/71),喉癌颈转移淋巴结组为22.7％(5/22),喉癌前病变组为11.8％(2/17),声带息肉组为6.7％(2/30),癌周正常喉组织为0％(0/20)。HPV DNA型别分布在喉癌中以HPV16、18型为主,喉良性病变中以HPV6、11型为主。表明喉癌发生发展与HPV感染相关。     

宫颈癌组织HPV16DNA相关序列检测与临床分析

用Southern blot核酸杂交技术检测宫颈组织HPV 16 DNA相关序列。山东省儿6例宫颈癌病人的癌组织HPV 16 DNA相关序列阳性率为50.9％,36例无宫颈疾病妇女的宫颈组织阳性率为5.6％,宫颈癌与HPV 16感染有很强的联系,OR=17.60,x~2=23.47,P<0.001.HPV 16 DNA相关序列阳性率与宫颈癌病人的年龄、肿瘤的组织类型、临床类型及临床分期均无统计学联系(P均大于0.25).本文首次从山东省宫颈癌病人癌组织中检测到HPV 33 DNA4例及HPV31 DNA 1例。     

宫颈癌组织中HPV16E6序列多态性及同源性分析

目的：探讨广东地区宫颈癌组织中HPV16肿瘤相关性抗原E6基因序列的多态性及同源性。方法：采用通用引物PCR直接测序法对宫颈癌标本中的HPV分型，从舍有HPV16型的标本中采用自行设计的多重引物通过巢式PCR扩增出HPV16E6，经DNA序列测定法检测其基因变异，进而分析其同源性。结果：50例宫颈癌组织HPV-DNA的检出率为78％．其中HPV16和HPV18型混合感染18例，单纯HPV16型感染15例。含有HPV16型的标本34例中扩增出HPV16E 625例。其中178位核苷酸变异较大，变异率为72％，其相应氨基酸均由天冬氨酸变为谷氨酸。结论：HPV16E6 DNA序列发生碱基替换的区域主要在氨基端94～241位，羧基端相对保守，未见变异。广东地区宫颈癌组织中HPV16E6的热点突变为Nt178。     

食管鳞癌组织HPV的检测及其意义的研究

目的:探讨人乳头状瘤病毒(HPV)与食管鳞癌的相关性,进一步明确人乳头瘤痛毒感染在食管癌病因学中的作用.方法:采用可检测23种HPV基因型的基因芯片和实时荧光定量PCR检测方法检测140例新鲜食管鳞癌组织的HPV型别,同时对照检测85例宫颈鳞癌组织中HPV的感染率.结果:85例宫颈鳞癌组织中HPV的阳性率为95.29%(81/85),共检测到9种HPV基因型,分别为HPV16、18、45、33、58、59、73、31和56,均为高危型感染.其中HPV16最常见,检出率达72.9%(62/85),其次是HPV18为16.5%(14/85),其他7型占28.2%(24/85),HPV双重感染检出率为11.8%(10/85).而在140例食管鳞癌组织中,未检测到任何基因型的HPV.结论:HPV感染似乎可能与高发区食管鳞癌的发生无关.     

声带息肉、喉乳头状瘤及喉癌组织内核仁组成区相关嗜银蛋白的定量学研究

采用核仁组成区嗜银蛋白（ＡｇＮＯＲｓ）银染技术对声带息肉、喉乳头状瘤及喉鳞状细胞癌各１０例进行银染制片。并采用电子计算机图象分析系统对ＡｇＮＯＲｓ数目、细胞核面积、颗粒形状团子平均值、颗粒面积及银染面积占核面积的比值５项参数进行定量学观察。结果显示，在喉癌组织中除颗粒形状因子低于喉乳头状瘤和声带息肉外，其它４项参数均明显高于喉乳头状瘤和声带息肉。本研究表明ＡｇＮＯＲｓ数目增高和形态改变在判断喉部良、恶性肿瘤方面具有一定的价值。     

人乳头瘤病毒DNA6/11、16、18在成人喉乳头状瘤中的表达探讨

为探讨乳头瘤病毒在成人喉乳头状瘤中的表达变化，采用原位杂交技术，用荧光素标记的人乳头瘤病毒（HPV）6／11、16、18型探针，对36例福尔马林固定，石蜡包埋的喉乳头状瘤组织进行HPV6／11、16和18型检测。结果：36例喉乳头状瘤中21例HPV6B／ll阳性；3例IIPV16阳性；HPV18无一例阳性，且阳性细胞主要分布于乳头浅层。结果提示喉乳头状瘤的发生与IIPV6／ll和16型有关，乳头浅层可能是HPV的重要作用部位。     

Human Papillomavirus DNA in Squamous Cell Carcinomas of the Respiratory and Upper Digestive Tracts

The prevalence of human papillomavirus (HPV) types 16 and 18in clinical samples of squamous cell carcinomas from respiratoryand upper digestive tracts was studied. HPV DNA of types 16and/or 18 was detected using the polymerase chain reaction (PCR)method in 16 out of 121 cases (13.2%). By Southern blot hybridization,however, only the DNA from a laryngeal and a tonsillar carcinomawas found to hybridize with the whole HPV 16 DNA probe (twoout of 16 HPV DNA-positive cases by PCR, 12.5%). None of theDNAs hybridized with the whole HPV 18 DNA probe. The discrepancyin the results of PCR and Southern blot hybridization methodsseemed to reflect their sensitivity. The possible relation betweenprevalence of HPV DNA and carcinogenesis in respiratory andupper digestive tract is discussed.     

采用原位杂交法检测生殖系统尖锐湿疣和鳞状细胞癌内乳头瘤病毒的感染

目的　探讨HPV感染与生殖系统尖锐湿疣和鳞状细胞癌的关系。方法　采用HPV6 /11、16 /18原位杂交试剂盒 ,对 5 5例生殖系统不同病变中乳头瘤病毒感染状况进行分析检测。结果　 2 0例生殖系统尖锐湿疣 19例阳性 ,其中 17例为HPV6 /11型 ,2例为HPV16 /18型。 2 0例生殖系统鳞状细胞癌中 ,15例阳性 ,均为HPV16 /18型。 10例正常生殖系统鳞状上皮组织和 5例外阴白斑组织均呈阴性。结论　HPV6 /11多与生殖系统良性疣状病变有关 ,而HPV16 /18感染多见于生殖系统恶性病变。     

Comparison of DNA Copy Numbers in Original Oral Squamous Cell Carcinomas and Corresponding Cell Lines by Comparative Genomic Hybridization

We analyzed regional DNA copy numbers in 4 oral squamous cell carcinomas (SCCs) by using comparative genomic hybridization, and compared them with those in cell lines derived from the SCCs. In the original tumors, DNA copy number increases were observed on chromosomes 5p (4/4 cases), 8q (4/4), 20p (3/4), 3q (2/4), 5q (2/4), 7p (2/4), 7q (2/4), 1ip (2/4), 11q (2/4) and 13q (2/ 4). Although most of these changes have been described previously for SCC tumors in the head and neck, the incidence of increases in 8q and 20p was much higher in the present study; this may be important in relation to cell line establishment, since 8q contains c- myc , which is involved in immortalization. No common chromosomal region with DNA copy number decreases was observed, except for 18q (2/4). When the original tumors and the cell lines were compared, their profiles were essentially similar with one exception. Further, there was no region that commonly changed in the cell lines, but not in the original tumors, suggesting that the DNA copy number changes observed in the cell lines mostly represent those of the original tumors.     

HPV Detection and Genotyping in Vulvar Squamous Cell Carcinoma in Northern Thailand

Background: The study was aimed to evaluate the prevalence and genotype distribution of HPV infection invulvar squamous cell carcinoma (SCC) in northern Thailand and the clinicopathological difference with regardto HPV infection status. Materials and Methods: Formalin-fixed paraffin-embedded tissue samples of vulvarSCC diagnosed between January 2006 and December 2012 were collected. HPV infection was detected by nestedpolymerase chain reaction (PCR) with primers MY09/11 and GP5+/6+. HPV genotyping was performed usingthe Linear Array Genotyping Test, followed by type-specific PCR targeting the E6/E7 region of HPV16/18/52if the Linear Array test was negative. The histologic slides of vulvar lesions and the medical records werereviewed. Results: There were 47 cases of vulvar SCC included in the study (mean patient age 57.9±13.2 years).HPV infection was detected in 29 cases (62%), all of which had single HPV infections. HPV16 accounted for 23(49%). The patients with HPV-positive SCC had a significantly younger mean age than those with HPV-negativetumors (52.7 years vs 66.2 years, p<0.001). There was no significant difference in tumor stage distribution withregard to the status of HPV infection. The presence of vulvar intraepithelial neoplasia (VIN) of usual type(basaloid or warty) was significantly more frequent in HPV-positive cases compared with HPV-negative cases(62% vs 6%, p<0.001), whereas differentiated-type VIN was more common in HPV-negative cases (24% vs0%, p=0.019). Conclusions: HPV infection was detected in 62% of vulvar SCC in northern Thailand. HPV16was the predominant genotype similar to the data reported from other regions. HPV-positive SCC occurredin younger patients compared with HPV-negative SCC, and was associated with usual-type VIN. Vaccinationagainst HPV16/18 may potentially prevent almost one half of vulvar SCC in northern Thailand.     

宫颈癌组织中HPV_(16)DNA基因同源序列的检测

206例不同病理诊断的宫颈脱落细胞和宫颈组织DNA,同探针杂交,检测HPV_(16)DNA同源性序列。结果表明:HPV_(16)DNA探针同宫颈癌脱落细胞杂交(65.2％)和同宫颈癌组织DNA杂交(66.7％),其杂交率无显著差别(P>0.05)。宫颈炎脱落细胞DNA和宫颈炎组织DNA与HPV_(16)DNA探针杂交率间的差异也无统计学意义。然而,宫颈癌和宫颈炎之间无论是脱落细胞或是组织DNA同探针杂交有显著差异(前者为65.2—66.7％,后者为26.1—28.5％)P<0.01。宫颈癌和宫颈炎DNA经酶切电泳后,Southern blot杂交结果提示HPV_(16)DNA整合在癌组织基因组中。     

喉癌细胞PCNA表达和DNA含量与临床预后的关系

 目的　探讨增殖细胞核抗原(PCNA)的表达和DNA含量对喉癌临床生物学特征及预后的关系。方法　用免疫组化SP方法和自动图像分析仪检测正常喉粘膜、喉鳞癌的PCNA和DNA指数(DI)，结合临床随访资料进行分析。结果　喉鳞癌PCNA的阳性表达率为100%，与正常喉组织有显著差异(P<0.01)，PCNA阳性表达强度和DNA指数随肿瘤分化程度的降低而相应增高(P<0.05)。两指标较高者生存时间较短，预后较差(P<0.05)。结论　PCNA的表达和DNA指数可为判断肿瘤的恶性程度及临床预后提供参考依据。     

Detection of High-Risk Human Papillomavirus Genotypes 16 and 18 in Head and Neck Squamous Cell Carcinomas in Jordan

Background: Recently, associations of the human papillomavirus (HPV) with head and neck cancer have become well established. Of particular concern, the severity and pathological outcomes of squamous cell carcinomas are remarkably affected by the genotypes of HPV present in such lesions. This study was conducted to investigate the occurrence of HPV genotypes, particularly high risk 16 and 18, among oral and laryngeal squamous cell carcinomas in Jordan. Methods: During the period of May 2015 to March 2016, we evaluated a total of 108 paraffin-embedded tissue samples, histologically confirmed as SCC, of both oral and laryngeal tumors for the presence of HPV DNA. DNA was extracted using a Zymogen commercial kit. HPV genotypes were detected by nested PCR using consensus primers followed by primer-specific PCR for HPV-16 and HPV-18 genotypes. The genotypes were confirmed by DNA sequencing methods. Results: Sixteen samples were positive for HPV DNA (14.8%) with higher rates in oral tumors compared to their laryngeal counterparts (20% and 6% respectively). The HPV-16 genotype predominated, being detected in 81.3% of the cases as a single infection and in 18.7% in combination with HPV-18. A significant association between the anatomical location and the HPV-16 genotype was observed (p < 0.05). In contrast, no significant associations could be established with tumor grade and gender or age. Conclusions: A relatively high rate of high-risk HPV genotypes, especially HPV 16, is evident in head and neck cancers SCCs in Jordan. Genotyping of HPV might be of considerable value for evaluation of progression.     

食管癌细胞株Eca109中人乳头瘤病毒(HPV)DNA的检测

应用分子生物学核酸杂交技术,以人乳头瘤病毒HPV6及HPV11型混合探针对食管ECA109鳞癌细胞株进行DNA斑点杂交。检测结果为阳性。为HPV与食管癌关系的研究进一步提供了证据。