用核酸杂交法研究人类乳头瘤病毒与口腔癌的相关性

以放射性核素￣（３２）Ｐ标记ＨＰＶ１１、ＨＰＶ１６、Ｅ＿６和ＨＰＶ１８ＤＮＡ探针，用核酸斑点杂交法，检测了部分口腔恶性肿瘤组织中的ＨＰＶ相关序列。（１）用ＨＰＶ１１探针检测１２例，其中口腔粘膜鳞状细胞癌（ｓｑｕａｍｏｕｓｃｅｌｌｃａｒｃｉｎｏｍａ，ＳＣＣ）８例，涎腺腺样囊性癌（ａｄｅｎｏｉｄｃｙｓｔｉｃｃａｒｃｉｎｏｍａ，ＡＣＣ）２例，涎腺多形性腺瘤恶变（ｍａｌｉｇｎａｎｔｐｌｅｏｍｏｒｐｈｉｃａｄｅｎｏｎｉａ，ＭＰＡ）１例，软组织胚胎性横纹肌肉瘤（ｅｍｂｒｙｏｉｄｒｈａｂｄＯｍｖｏｓａｒｃｏｍａ，ＥＲＳ）１例；结果阳性５例（５／１２），弱阳性１例，６例为阴性。（２）用ＨＰＶ１６Ｅ＿６探针检测１０例，其中ＳＣＣ６例，ＡＣＣ２例，ＭＰＡ１例，ＥＲＳ１例；结果阳性６例（６／１０），弱阳性１例，３例为阴性。（３）用ＮＰＶ１８探针检测１２例，肿瘤类型及例数同ＨＰＶ１１检测组，结果阳性３例（３／１２），阴性９例（９／１２），非肿瘤对照组共４例（唇、腭裂各２例），除１例唇裂出现了与ＨＰＶ１１和ＨＰＶ１６Ｅ＿６杂交呈弱阳性外，余均为阴性。     

口腔鳞癌中人乳头瘤病毒的研究

为了探讨人乳头瘤病毒（ｈｕｍａｎｐａｐｉｌｏｍａｖｉｒｕｓ，ＨＰＶ）１６、１８型及其与癌的关系，作者采用聚合酶链反应（ｐｏｌｙｍｅｒａｓｅｃｈａｉｎｒｅａｃｔｉｏｎ，ＰＣＲ）技术，检测２３例口腔鳞状细胞癌（ｏｒａｌｓｑｕａｍｏｕｓｃｅｌｃａｒｃｉｎｏｍａ，ＯＳＣＣ），正常口腔粘膜（ｎｏｒｍａｌｏｒａｌｍｕｃｏｓａ，ＮＯＭ）中的ＨＰＶ１６型和１８型ＤＮＡ中的Ｅ６、Ｅ７基因片段，并对其ＰＣＲ产物进行Ｓｏｕｔｈｅｒｎ印迹杂交分析，以研究ＨＰＶ与ＯＳＣＣ的关系。结果显示：ＯＳＣＣ中ＨＰＶＤＮＡ阳性率４７．８％（１１／２３），其中ＨＰＶ１６型６例，ＨＰＶ８型３例，ＨＰＶ１６和１８型复合感染２例，ＮＯＭ中ＨＰＶ阳性率２０％（２／１０），２例阳性者，均为ＨＰＶ１６型。研究结果提示：高危型ＨＰＶ与口腔鳞癌可能有关，二者的确切关系尚需进一步研究。但ＨＰＶ致癌基因Ｅ６、Ｅ７片段在口腔鳞癌中的检出，为鳞癌的病因研究开辟了新的前景     

多聚酶链反应研究人乳头瘤病毒感染和口腔鳞状细…

建立了一种简并引物ＰＣＲ技术扩增多型人乳头瘤病毒（ＨＰＶ）ＤＮＡ的方法，本方法至少可以特异地扩增ＨＰＶ６，１１，１６，１８及３３。将其用于扩增福尔马林固定石蜡包埋口腔鳞状细胞癌及癌变白斑组织中的ＨＰＶｓＤＮＡ，其阳性例数分别为２２／３０，２３／３０例。本研究表明ＨＰＶ极可能参与了口腔鳞状细胞癌和癌变白斑的癌发生。     

多聚酶链反应研究人乳头瘤病毒感染和口腔鳞状细胞癌及癌变白斑的关系

建立了一种简并引物ＰＣＲ技术扩增多型人乳头瘤病毒（ＨＰＶ）ＤＮＡ的方法，本方法至少可以特异地扩增ＨＰＶ６，１１，１６，１８及３３。将其用于扩增福尔马林固定石蜡包埋口腔鳞状细胞癌及癌变白斑组织中的ＨＰＶｓＤＮＡ，其阳性例数分别为２２／３０，２３／３０例。本研究表明ＨＰＶ极可能参与了口腔鳞状细胞癌和癌变白斑的癌发生。     

人乳头瘤病毒16型在口腔癌及癌旁组织中的存在状态

采用斑点杂交和Ｓｏｕｔｈｅｒｎ印迹杂交检测２５例口腔鳞癌及其相应癌旁组织中的人乳头瘤病毒（ＨＰＶ）１６型ＤＮＡ，口腔癌ＨＰＶ１６ＤＮＡ的阳性率为２４．０％（６／２５），癌旁组织为８．０％（２／２５）。经ＢａｍＨＩ、ＰｓｔＩ酶切杂交后出现的阳性区带显示ＨＰＶ１６ＤＮＡ在口腔癌及癌旁组织中均以游离状态存在。作者推测ＨＰＶ１６ＤＮＡ在口腔癌的致癌机理中可能作为一个协作因子发生作用。     

口腔颌面部病变组织中人乳头瘤病毒DNA的检测

为探讨人乳头瘤病毒（ＨＰＶ）和口腔颌面部肿瘤以及某些口腔粘膜病的关系，应用聚合酶链反应技术（ＰＣＲ）检测４型人乳头瘤病毒６，１１，１６，１８型ＤＮＡ．结果：４５例口腔颌面部良性病变和恶性肿瘤患者，４８份组织标本中的人乳头瘤病毒ＤＮＡ（ＨＰＶ－ＤＮＡ），总阳性率达７５．６％（３４／４５）．其中恶性肿瘤ＨＰＶ－ＤＮＡ阳性率为７６．９（２０／２６）．良性病变ＨＰＶ－ＤＮＡ阳性率为７３．７％（１４／１９）．恶性肿瘤和良性病变２组间ＨＰＶ－ＤＮＡ的型别有显著的差异，恶性肿瘤以ＨＰＶ１６和ＨＰＶ１８型为主，良性病变以ＨＰＶ１８和ＨＰＶ６为主．非肿瘤对照组共３０人ＨＰＶ－ＤＮＡ阳性率为１０％，均为ＨＰＶ１８型．ＨＰＶ１８在口腔颌面部恶性肿瘤和良性病变组织中阳性率均较高，但两者间无统计学差异     

口腔鳞癌中HPV感染及其对p5 3改变影响的研究

目的：探讨高危型人乳头状瘤病毒（ＨＰＶ）在口腔鳞癌中的感染情况及其对Ｐ５３蛋白表达和ｐ５３突变的影响。方法：采用免疫组化和ＰＣＲ－ＳＳＣＰ方法，分别检测４０例来癌中高危型ＨＰＶＥ６蛋白表达、Ｐ５３蛋白表达和ｐ５３基因突变的情况。结果：９例ＨＰＶＥ６蛋白染色阳性，阳性率２２．５％（９／４０），与正常粘膜对照组有显著差异（Ｐ＝０．０２１）。ＨＰＶ阳性组中Ｐ５３蛋白表达率１１．１％（１／９），ＨＰＶ阴性     

口腔鳞状细胞癌与人乳头状瘤病毒感染的关系

本文用抗牛乳头状瘤病毒（ＢＰＶ）的抗血清对３１例口腔鳞状细胞癌中的人乳头状瘤病毒（ＨＰＶ）结构抗原进行检测，证明其中３０例口腔鳞状细胞癌中ＨＰＶ结构抗原一，说明口腔癌的发病与ＨＰＶ感染有密切关系，对防治ＨＰＶ感染，减少口腔癌的发病有着重要意义。     

人乳瘤病毒16型和EB病毒感染与口腔鳞癌关系

用多聚酶链反应技术为２１例口腔鳞癌及癌旁正常组织成对标本中的人乳头瘤病毒１６型（ＨＰＶ１６）和ＥＢ病毒（ＥＢＶ）ＤＮＡ作检测。结果口腔鳞癌和癌旁正常组织ＨＰＶ１６ＤＮＡ阳性率分别为５２．４％和１４．３％，ＥＢＶ为８５．７％和３３．３％，ＨＰＶ１６和ＥＢＶ均为阳性者分别为４７．６％和９．５％，癌组织和癌旁正常组织间统计学上均有显著性差异。提示口腔粘膜ＨＰＶ１６或ＥＢＶ感染均与口腔鳞癌发生有关。ＨＰＶ１６和ＥＢＶ在口腔粘膜感染可能有协同致口腔鳞癌作用。     

人类乳头瘤病毒在颌面部鳞状细胞癌感染的检测

用核酸分子原位杂交方法观察人类乳头病病毒ＨＰＶ６Ｂ．１１型病毒与颌面部鳞状细胞癌的关系，结果：病理诊断为鳞状细胞癌Ⅲ级１２例中，ＨＰＶ阳性者９１．７％。ＨＰＶ阴性者８．３％，鳞状细胞癌Ｉ级８例，ＨＰＶ阳性者占２５％，ＨＰＶ弱阳性占６２．５％，ＨＰＶ阴性占１２．５％，结果揭示了ＨＰＶ６Ｂ．１１型ＤＮＡ在口腔恶性鳞状细胞癌中有过表达，提示颌面部鳞状细胞癌的分化程度可能与ＨＰＶ病毒感染有密切关系。     

In situ DNA hybridization analysis of oral papillomas, leukoplakias, and carcinomas for human papillomavirus.

Twenty-one papillomas, 23 ordinary benign keratoses, 13 smokeless tobacco keratoses, 10 verrucous hyperplasias, 10 verrucous carcinomas, 17 squamous cell carcinomas, 3 epithelial dysplasias, and 6 lichen planus lesions were evaluated for human papillomavirus (HPV) types 6/11, 16/18, and 31/33/35, with biotinylated double-stranded DNA probes by in situ hybridization. Sixty-two percent (13/21) of oral squamous papillomas were positive for HPV DNA. HPV DNA types 6 and 11 demonstrated the strongest reactivity. Of the 13 cases, 10 also showed some reactivity with HPV-16/18 and -31/33/35. None of the cases of keratoses, epithelial dysplasia, squamous cell carcinoma, verrucous hyperplasia, verrucous carcinoma, or lichen planus were positive for HPV DNA. This study confirms the consistent and frequent finding of HPV DNA in oral squamous cell papillomas and the inconsistency of being able to identify HPV DNA in keratotic, premalignant, or cancerous lesions of the oral mucous membranes.     

Detection of human papillomavirus DNA in oral mucosal lesions using in situ DNA-hybridization applied on paraffin sections

The in situ DNA hybridization technique, carried out under stringent conditions, was used to detect human papillomavirus (HPV) DNA of types 6, 11, and 16 in paraffin sections of 32 surgically treated oral mucosal lesions. Expression of HPV structural proteins was analyzed by means of the immunoperoxidase (IP-PAP) method. A total of 10 (31.3%) of the 32 lesions proved to express HPV antigens, which were found in 4 of 7 squamous cell papillomas, in 2 of 2 classic condylomas, in 2 of 10 papillary hyperplasias, and in 2 of 3 leukoplakia lesions. Two of the squamous cell papillomas contained HPV 6 DNA, and 4 additional lesions were positive for HPV 11 DNA. In one of the condylomas, a double infection by HPV 6 and 11 was found, while the second was positive for HPV 11 alone. Both the HPV antigen-positive papillary hyperplasias contained HPV 6 DNA, as did the HPV antigen-positive leukoplakia lesions. Of the latter, one was infected by HPV 6 and 11. DNA of the "high-risk" HPV 16 was contained in two lesions: one lichen planus lesion and one of the two squamous cell carcinomas. The results confirm the previously reported evidence of HPV involvement in oral mucosal lesions. The implications of these findings are discussed in terms of the well-established premalignant character of oral leukoplakia and oral lichen planus, although far less commonly versus leukoplakia, with special emphasis on the discovery of the "high-risk" HPV 16 in the latter as well as in oral cancer.(ABSTRACT TRUNCATED AT 250 WORDS)     

人乳头状瘤病毒感染与儿童口腔黏膜良性上皮增生

目的 探讨儿童口腔黏膜良性上皮增生性病损与人乳头状瘤病毒(human papillomavirus，HPV)感染及其类型的关系。方法 选取四川大学华西口腔医学院病理科近10年30例儿童口腔黏膜良性上皮增生性病损的病例，复习其临床病理特征及切片，并采用免疫组织化学及原位杂交方法检测HPV共同抗原及其类型。结果 口腔鳞状细胞乳头状瘤(squamous cell papilloma，SCP)20例(66．7％)，尖锐湿疣(condyloma acuminatum，CA)6例(20．0％)，口腔黏膜局灶性上皮增生(focal epithelial hyperplasia，FEH)4例(13．3％)。HPV检测结果显示：HPV共同抗原阳性者占73．3％(22／30)，其中SCP占75．0％(15／20)，6例CA HPV共同抗原均为阳性，4例FEH中仅1例HPV共同抗原为阳性；HPV类型以高危型HPV16／18为主，占77．3％(17／22)，其次是HPV6和HPV11。结论 儿童口腔黏膜良性上皮增生性病损与HPV感染关系密切，病毒类型以高危型HPV16／18为主，其病毒类型是否与成人(以HPV6、11为主)不同尚待进一步研究。     

Oral squamous papillomas: detection of HPV DNA by in situ hybridization

Oral squamous papillomas were segregated from other papillary lesions on the basis of histopathologic features. Twenty representative papillomas were evaluated for the presence of papillomavirus genus-specific antigen with the use of an immunoperoxidase technique. These same tumors were analyzed for human papillomavirus (HPV) types 2, 4, 6, and 11 with biotinylated full-length double-stranded DNA probes by in situ hybridization. Only one case exhibited papillomavirus antigen reactivity. Alternatively, seven of twenty cases (35%) yielded positive results for HPV 6 or 11 DNA; one papilloma exhibited a dual infection with both HPV 2 and 6 when assayed under conditions of high-stringency hybridization. It is concluded that some oral squamous papillomas harbor HPV genotypes akin to those encountered in genital tract condylomas. Viral DNA can be detected in the absence of capsid antigen immunoreactivity, thereby obviating the use of antigen detection assays for determining the presence or absence of virus.     

Biopsy vs. superficial scraping: detection of human papillomavirus 6, 11, 16, and 18 in potentially malignant and malignant oral lesions

BACKGROUND: Several epidemiologic studies have shown a broad variation in the prevalence of human papillomavirus (HPV) in oral precancerous tissues and oral carcinomas. METHODS: Biopsies and superficial scrapes of lesions, clinically suspected of HPV infection, were taken from patients with potentially malignant and malignant oral lesions, and subject to HPV DNA detection by PCR-Southern blot analysis. RESULTS: From 22 patients with potentially malignant and malignant lesions analyzed, 41% of the biopsies were HPV DNA positive, whereas 95-100% of the superficial scrapes were positive (McNemar, P < 0.0001). Clinical presumption of HPV infection detected 67% (P < 0.0001) of the HPV DNA positive cases compared with 48% (P < 0.0001) determined by cytology and histopathology. The prevalence of HPV 6, 11, 16 and 18 in the oral mucosa was studied in 59 individuals. While 9% of normal controls were HPV DNA positive, 100% of the patients with potentially malignant and malignant lesions were HPV DNA positive, and the prevailing genotype was HPV 16 followed by HPV 18. CONCLUSIONS: The higher HPV DNA detection rate in superficial oral scrapes than in biopsies suggests that accurate epidemiological information on oral HPV infection/oral carcinogenesis depends not only on the DNA detection technique, but also on the tissue/cell sampling procedure.     

In situ detection of human papillomavirus Types 13 and 32 in focal epithelial hyperplasia of the oral mucosa

17 cases of focal epithelial hyperplasia of the oral mucosa (FEH, Heck's disease) were investigated for the presence of human papillomavirus (HPV) nucleic acid sequences by means of in situ DNA hybridization using biotinylated DNA probes of HPV types 1, 6, 11, 13, 16, 18, and 32. Ten of 17 cases were positive for HPV 13 DNA in contrast to 6 of 17 positive cases obtained after application of the HPV 32 probe, with a double infection in one case. The results of our study suggest, that HPV 13 and HPV 32 are very specifically found in lesions of FEH and can be detected in a high percentage of cases using in situ hybridization.     

艾滋病毒感染患者口腔黏膜疣状肿块人类乳头状瘤病毒感染的分型研究

目的探讨艾滋病毒感染患者口腔戮膜疵状肿块人类乳头状瘤病毒(1-IPV)感染及感染类型。方法应用 HPV Ll区通用引物Gp5十//G p6+和1-IPV特型引物(HPV6/II,16,18,31,33),通过聚合酶链反应(PCR),对34例艾滋病毒感染患者口腔豁膜沈状肿块人类乳头状瘤病毒(HPV) DNA进行检测。结果艾滋病毒感染患者口腔豁膜优状肿块中,HPV感染率为88.2%;亚型HPV6/II感染率为47.06%, HPV16感染率为II.76%, 1U〕V 18感染率为 2.94 % , HPV31感染率为5.88%。结论艾滋病毒感染患者口腔庆状肿块和H PV感染关系密切,而且大多数和低危型HPV6/II感染有关,少数病例和高危型HPV16,18,31感染有关;同一病例可以感染两种以上HPV亚型。     

Analysis of HPV 16 and 18 by in situ hybridization in oral papilloma of HIV+ patients

Human immunodeficiency virus (HIV) infection facilitates the development of other infections and lesions including oral papilloma, which has been associated with human papillomavirus (HPV). In analyzing the presence of HPV 16 and 18 by in situ hybridization in oral papillomas from five HIV+ male dental patients, HPV 16 and 18 were observed in 9 of 16 (52.2%) histopathologic specimens. All positive lesions occurred in heterosexual males admitting to oral sexual contact with a female partner (Pearson's correlation; p = 0.0088). These results suggest that oral sexual behavior may be a contributing factor in the presence of HPV 16 and 18 in oral papilloma.     

In situ detection of HPV DNA in oral mucosal lesions. A comparison of two hybridization kits

The purpose of this study was to compare the sensitivity of ViraType in situ hybridization kit (Life Technologies, Inc. [LT] and PathoGene (Enzo Diagnostics, Inc. [ED]) in situ hybridization kit for human papillomavirus (HPV) DNA detection in oral tissue. Forty benign oral lesions histologically suspicious for HPV infection were analyzed. Specimens were hybridized with DNA probes specific for HPV types 6/11, 16/18, and 31/33/35 [LT] and HPV types 6/11, 16/18, and 31/33/51 [ED]. Positive hybridization reactions were seen for HPV DNA type 6/11 only. Hybridization occurred significantly more often (p less than 0.01, McNemar Exact Test) in LT probed specimens (20/40) than ED assayed sections (12/40). HPV DNA sequences were found in 100% condyloma acuminata (13/13), 100% verruca vulgaris (4/4), and 13% squamous papilloma (3/23) using the LT system. The ED probes yielded positive signals in 77% condyloma acuminata (10/13), 25% verruca vulgaris (1/4), and 4.4% squamous papilloma (1/23). A more intense hybridization signal was exhibited using the LT system. The results indicate that the LT probes and detection reagents are more sensitive for detecting HPV DNA in oral mucosal specimens.     

Unusual HPV types in oral warts in association with HIV infection

Human papillomaviruses (HPV) are associated with certain oral soft tissue lesiona, such as papillomas, warts, condylomata, and focal epithelial hyperplasia (FEH). HPV types 2, 6, 11, 16, and 18 have been identified in some of these oral lesions, while HPV 13 and 32 are associated with FEH. Little is known about the HPV types in oral warts of persons infected with human immunodeficiency virus (HIV). In this study, oral warts in 17 HIV-seropositive individuals were biopsied. Southern blot analyses were performed and the HPV types found were HPV 7 (7/17), 13 (1/17), 32 (1/17), and 18 (1/17). The presence of HPV type 7 is unusual in that it normally is found only in butcher's warrs. There was no correlation between HPV type, histopathology, and clinical appearance of the lesions examined, except that the flat (FEH type) warts contained HPV types 13, 18 and 32 (1 of each). HIV infection appears to predispose individuals to oral infection with unusual HPV types.