膀胱砂状移行细胞癌中高危人乳头瘤病毒的检测

目的 地高危人乳头瘤病毒（ＨＰＶ）１６、１８ＤＮＡ在膀胱癌组织中进行定位研究。方法 运用地高辛标记的原位杂交技术对５２例膀胱砂状移行细胞癌中高危ＨＰＶＤＮＡ进行检测。结果 ＨＰＶＤＮＡ的阳性信号存在于肿瘤细胞核内，呈占 或点片状，其中以点状为主，经８９．５％。癌旁不典型增生上皮、癌旁正常的上皮组织及Ｂｒｕｎｎ巢可同时有高危ＨＰＶ的感染，但表达呈点片状。５２例胱乳头状移行细胞癌中高危型ＨＰＶ１６、１     

膀胱乳头状移行细胞癌中高危人乳头瘤病毒的检测

目的对高危人乳头瘤病毒（ＨＰＶ）１６、１８ＤＮＡ在膀胱癌组织中进行定位研究。方法运用地高辛标记的原位杂交技术对５２例膀胱乳头状移行细胞癌中高危ＨＰＶＤＮＡ进行检测。结果ＨＰＶＤＮＡ的阳性信号存在于肿瘤细胞核内,呈点状或点片状,其中以点状为主,约８９５％。癌旁不典型增生上皮、癌旁正常的上皮组织及Ｂｒｕｎｎ巢可同时有高危ＨＰＶ的感染,但表达呈点片状。５２例膀胱乳头状移行细胞癌中高危型ＨＰＶ１６、１８ＤＮＡ阳性１９例,阳性率为３６５％;ＰＴａＴ２期１７例,ＰＴ３Ｔ４期２例;Ｇ１,２级１４例,Ｇ３级５例。随着肿瘤分期分级的增加,ＨＰＶ１６及ＨＰＶ１８的感染率有逐渐降低的趋势,但差异无统计学意义（Ｐ＞００５）。结论病毒ＤＮＡ在膀胱癌变组织、癌旁正常及不典型增生组织中均有不同程度的表达。膀胱乳头状移行细胞癌ＨＰＶ感染率较高,浸润较浅分化较好的肿瘤更多见,提示该病毒感染可能是膀胱癌发生的早期诱因之一。     

P53,Ras基因突变及HPV感染与膀胱癌的关系

应用分子生物学方法检测３２例膀胱移行上皮癌中Ｐ５３、Ｋ－ｒａｓ、ＨＰＶ基因，发现有３４．４％的膀胱癌Ｐ５３基因突变，２１．９％膀胱癌Ｋ－ｒａｓ基因突变，６．３％膀胱癌ＨＰＶ阳性。说明膀胱移行上皮癌的发生与Ｐ５３、Ｒａｓ基因突变及ＨＰＶ感染有关。膀胱癌的发生是多种基因的变化通过多种途径引起的。     

人膀胱移行细胞癌细胞系PUMC-91细胞HPV-18感染

为探讨高危型人乳头状瘤病毒(ＨＰＶ 16 /18)与膀胱癌的关系 ,我们对膀胱移行细胞癌细胞系ＰＵＭＣ 91中的ＨＰＶ感染情况进行了研究 ,报告如下。材料和方法　细胞培养 :人膀胱移行细胞癌ＰＵＭＣ 91细胞系及宫颈癌ＨｅＬａ细胞系 37℃闭式培养于含 10 %新生牛血清的ＲＰＭＩ 16 40培养液中。ＤＡ的提取 :传代培养收集 1× 10 6～ 1×10 7细胞 ,常规酚 氯仿抽提ＤＮＡ ,测量吸光度Ａ值 ,贮存于 - 2 0℃。ＨＰＶＤＮＡ探针的分离和提纯 :用限制性内切酶ＥｃｏＲＩ消化含ＨＰＶ 18 ＰＢＲ 32 2质粒 ,过夜消化 ,0 .8%琼脂糖凝胶电泳 ,40Ｖ ,3ｈ…     

人类乳头状瘤病毒与腺性膀胱炎的相关性研究

人类乳头状瘤病毒 (ＨＰＶ)是一类与泌尿生殖系肿瘤关系密切的ＤＮＡ病毒。本研究试图通过免疫组织化学方法检测腺性膀胱炎等病变标本的ＨＰＶ感染情况 ,揭示ＨＰＶ与腺性膀胱炎的相关性 ,为探讨腺性膀胱炎的病因提供理论依据。1　材料与方法1 .1 　 标本来源36例腺性膀胱炎石蜡标本、2 6例膀胱移行细胞癌并腺性膀胱炎石蜡标本、2 6例膀胱移行细胞癌石蜡标本、2 5例正常膀胱石蜡标本 (生前无泌尿系疾病症状的成人尸检膀胱组织石蜡标本 )均来自广西医科大学附属第一医院病理科 ,均经病理检查确诊。鼠抗ＨＰＶ单克隆抗体 (ＨＰＶ通用型 )免疫…     

高危性人乳头瘤病毒与抑癌基因P53在膀胱癌中的表达

用ＰＣＲ法检测５２例膀胱癌组织中人高危性乳头瘤病毒（ＨＰＶ）ＤＮＡ，同时用免疫组化方法检测其Ｐ５３蛋白的过度表达，发现２８例（５３．８４％）ＨＰＶＤＮＡ阳性，１９例（３６．５４％）肿瘤细胞核Ｐ５３染色阳性，二者同时阳性者３例（５．７７％）。ＨＰＶＤＮＡ阳性者多见于高分化、非侵袭性肿瘤中，相反Ｐ５３蛋白阳性则主要表现在低分化、侵袭性肿瘤中。ＨＰＶＤＮＡ和Ｐ５３的表达呈显著性负相关（ｒ＝－０．５７６９）。结果提示：ＨＰＶ感染或Ｐ５３的过度表达与膀胱肿瘤的生物学行为有关，并可将其作为膀胱肿瘤的预后评价指标。     

膀胱癌组织中血管内皮生长因子的表达及与血管生成的关系

目的 探讨血管内皮生长因子（ＶＥＧＦ）在原发性膀胱移行细胞癌中的表达及其与膀胱癌血管生成之间的关系。方法 免疫组织化学技术检测６８例原发性膀胱移行细胞癌和７例正常膀胱组织中血管内皮生长因子的表达,测定４０例浸润性膀胱癌的微血管密度,并分析ＶＥＧＦ和ＭＶＤ间以及它们与膀胱癌病理分级和临床分期间的关系。结果 ＶＥＧＦ在正常膀胱中不表达或低表达,而在膀胱表达较强;ＶＥＧＦ表达及ＭＶＤ值均与肿瘤的病理分级     

膀胱移行细胞癌VEGF表达及与血管形成定量的关系

为了探讨膀胱移行细胞癌中血管内皮生长因子（ＶＥＧＦ）表达及其与血管形成定量关系，应用免疫组织化学方法，对６２例原发性膀胱移行细胞癌及８例正常膀胱组织中ＶＥＧＦ进行检测，并对其在３０例浸润性膀胱癌组织中表达与血管形成定量关系进行了研究。结果发现正常膀胱组织均为阴性反应，膀胱癌组织中ＶＥＧＦ蛋白阳性表达率为５６％。低分化和浸润性癌中ＶＥＧＦ蛋白阳性表达率明显高于高分化和表浅性癌组（Ｐ＜００５），浸润性膀胱癌组织中血管形成定量与ＶＥＧＦ表达密切相关（Ｐ＜００１）。结果提示：ＶＥＧＦ表达对膀胱癌生物学行为有重要影响。ＶＥＧＦ是膀胱癌发生发展过程中一个主要血管生成因子，ＶＥＧＦ蛋白表达和血管形成定量有可能成为预测膀胱癌转移和预后的指标     

用原位杂交检测膀胱癌中人乳头瘤病毒DNA

目的：研究多块活检及多探针原位杂交系统能否增加人乳头瘤病毒（ＨＰＶ）阳性检出率,探讨循环中的抗－ＨＰＶ抗体与ＨＰＶ阳性膀胱癌之间及ＨＰＶＤＮＡ存在与肿瘤分级、分期及进展等临床病理学表现之间的可能关系。方法：用酶联免疫吸附试验检测４３例膀胱移行细胞乳头状癌患者血中抗人及抗牛抗体。仅有１份肿瘤标本作组织病理学研究者２１例,有２～９份标本者２２例,所有标本均有１０％福尔马林固定、石蜡包埋、切片。原位杂交用２种普通广谱ＨＰＶＤＮＡ探针（ＨＰＶ染色盒,Ｋ６０２、Ｄａｋｏ和ＨＰＶ－Ｏｍｎｉｐｒｏｂｅ,Ｄｉｇ…     

血管生成素-2在膀胱移行细胞癌中的表达及意义

目的　初步探讨血管生成素 2 (Ang 2 )在膀胱移行细胞癌组织中的表达及其与临床分期、病理分级的关系。方法　应用免疫组织化学S P法检测 4 3例初治膀胱癌及 2 8例正常膀胱组织中的Ang 2表达水平 ,并与临床资料对照进行统计分析。 结果　正常膀胱组织中未见Ang 2阳性染色 ;4 3例膀胱移行细胞癌中Ang 2阳性染色者 2 1例 ,Ang 2在许多膀胱癌细胞和癌组织中微血管内皮上呈强阳性染色 ,且染色率随膀胱癌病理分级、临床分期的上升而升高 (P <0 .0 5 )。结论　①Ang 2促进肿瘤新生血管形成 ,参与膀胱癌的发生和发展。②Ang 2的表达与膀胱移行细胞癌的临床分期、病理分级正相关。     

膀胱癌患者阴毛毛囊人乳头瘤状病毒的检测分析

目的人类乳头瘤状病毒（human papillomavirus，HPV）与膀胱癌关系密切，HPV在膀胱癌组织中的检出率很高，但其感染途径尚不清楚，本文旨在明确膀胱癌患者阴毛毛囊HPV的感染情况。方法PCR体外扩增和DNA反向点杂交相结合的DNA芯片技术测定19例膀胱尿路上皮细胞癌患者和30例健康人阴毛毛囊HPV感染情况。结果19例膀胱尿路上皮细胞癌患者的阴毛毛囊有2例发现HPV感染，HPV感染的亚型为HPV6和HPV18，HPV的感染率为10．5％；30例健康人中2例阳性，其亚型均为HPV6，感染率为6．7％。结论膀胱尿路上皮细胞癌患者阴毛毛囊不是膀胱癌组织中HPV的感染途径（P〉0．05），推测膀胱癌组织HPV感染的途径可能为通过尿道逆行感染。     

胃癌及癌旁组织中HPV16感染与p21基因失活的相关研究

目的 探讨胃癌及癌旁组织中HPV16的感染及p21基因突变是否存在协同致癌作用及其与胃癌患者预后的关系。方法 采用多聚酶链反应技术检测46例胃癌及癌旁组织标本中HPV16型感染,并采用免疫组织化学链霉菌抗生物素蛋白－过氧化酶连接法对胃癌p21基因突变进行检测。结果 HPV16阳性率为41．30％（19／46）,p21阳性表达为52．17％（24／46）。随访表明46例有21例出现复发或远处转移,其中HPV16阳性者复发率为73．68％（14／19）,p21基因阳性表达者复发率为66．60％（16／24）。结论 HPV16感染可能是胃癌发生的病因之一,而p21基因失活具有协同致癌作用,亦可作为判断胃癌预后的指标之一。     

Human papillomavirus sequences are not detectable by Southern blotting or general primer-mediated polymerase chain reaction in transitional cell tumours of the bladder

Summary A large series of transitional cell tumours has been screened for the presence of human papillomavirus (HPV) sequences using Southern blotting and general primer-mediated polymerase chain reaction (GP-PCR). The latter technique allows the detection of a broad spectrum of both sequenced and unsequenced HPV types using two primer pairs located in the highly conserved L1 and E1 regions of the HPV genome. No evidence for HPV infection was found in 100 transitional cell tumours, 6 cases of carcinoma in situ, 2 adenocarcinomas and a squamous carcinoma of the bladder and 3 cases of cystitis. Similarly, 12 bladder tumour cell lines were HPV-negative in these assays. Cervical carcinoma cell lines containing from 1–3 to 600 copies of the HPV genome were used as positive controls and were scored positive in all assays by both Southern blotting and GP-PCR. It is concluded that despite the close proximity of the urothelium to the genital mucosa and the resemblance of some bladder tumours to known HPV-induced lesions in other tissues, HPV infection is absent or very uncommon in bladder tumours.     

Human papillomavirus associated with bladder cancer.

Recently published data have suggested a link between active human papillomavirus (HPV) infection and the development of bladder cancer. This study was undertaken to test for HPV genomic material in the tumors of patients without evidence of ongoing viral infection. Twenty-three consecutive patients with clinical evidence of intravesical neoplasia and no history of HPV infection or clinical evidence of intercurrent disease, underwent cystopanendoscopy and biopsy as part of the routine evaluation and treatment of their tumor. Routine pathologic evaluation and southern blot analysis of biopsy material were done to establish the presence or absence of HPV DNA in the bladder tumors. Twenty-one tumors were identified by routine histology: 20 were low-to-moderate grade transitional cell carcinomas; 1 was found to be squamous cell carcinoma; 1 patient had moderate dysplasia; and 1 patient had evidence of inflammation. Four of the 20 transitional cell tumors (20%) were found to contain HPV DNA. In addition, the patients with dysplasia and cystitis were also shown to have HPV genomic material in their biopsy specimens. Viral types 6/11, 16/18, and 31/33 were found. The 20 percent incidence of HPV genomic material in bladder tumors from patients without clinical evidence of viral infection is in keeping with the observations of other investigators. We present the implication of these findings within the context of our current understanding of viral oncogenesis in the urinary bladder.     

Human papillomavirus and bladder cancer

Bladder cancer remains an important cause of oncological morbidity and mortality in women. Known etiological agents include smoking and exposure to certain industrial chemical compounds, though the origin of the majority of cases remains unknown. Human papillomavirus infection is also common in women and has been closely linked to the development of carcinoma of the cervix. It has been suggested that infection with HPV may also be an important factor in the subsequent development of bladder cancer. A number of studies using various techniques of molecular biology have looked at the relationship between HPV infection and bladder cancer. Although the results are somewhat conflicting, the overall picture would suggest little involvement of HPV in the evolution of bladder cancer, except possibly in a small group of patients who are immunocompromised.     

Human papilloma virus DNA and p53 mutation analysis on bladder washes in relation to clinical outcome of bladder cancer

OBJECTIVES: High-risk human papilloma virus (HPV) types stimulate degradation and deactivation of protein associated with the p53 tumour suppressor gene via the ubiquitin-dependent pathway. For a long time, changes of the p53 tumour suppressor gene have been correlated with poor clinical outcome in patients with superficial bladder cancer. We aimed to study the association between presence of (high-risk) HPV DNA, p53 status, and clinical outcome in bladder cancer patients. This study must be seen as a preliminary study to investigate this potentially important problem. MATERIAL AND METHODS: From 107 patients, 166 bladder wash samples were obtained. p53 status was determined by mutation analysis, HPV detection, and genotyping by the SPF(10)-LiPA assay. Clinical data were abstracted from the medical files. RESULTS: The prevalence of all-type and high-risk HPV infection in malignancies of the bladder was 15.2% and 8.1%, respectively. In high-grade tumours this prevalence was 18.2% and 10.6%, respectively. In grade 1, 2 and 3 tumours the infection rate of high-risk HPV types was 0%, 3.3%, and 10.6%, respectively (trend test: p=0.221). In Ta, T1, and T2-T4 tumours the high-risk HPV infection rate was 0%, 12.5% and 18.2%, respectively (trend test: p=0.045). In the p53 wild-type patients who showed progression, 1 of 9 patients had a high-risk type HPV infection. In the group of wild-type patients who showed no progression, 4 of 37 patients had a high-risk type HPV infection (odds ratio: 1.03; 95% confidence interval, 0.1-10.5). CONCLUSIONS: The data of this pilot study show the suggestion of a positive trend in the correlation between tumour grade/stage and high-risk type HPV infection. However, no additional risk for progression is found for p53 wild-type patients with a high-risk HPV infection.     

Association between urothelial carcinoma after renal transplantation and infection by human papillomavirus types 16 and 18

Objective

To explore the association between urothelial carcinoma following renal transplantation and infection by human papillomavirus (HPV) types 16 and 18.

Materials and Methods

Of 3780 patients who underwent renal transplantation, we identified 90 cases of urothelial carcinoma. Tumor tissues collected from the 90 renal transplant recipients were compared with those from 30 nontransplanted patients with bladder cancer (control group) for HPV types 16 and 18 using polymerase chain reactions.

Results

Seven transplanted patients were HPV positive: HPV-16 was detected in 3 patients with bladder cancer (3/90; 3.3%), and HPV-18 in 2 patients with bilateral pelvic ureteral carcinoma (2/90; 2.2%), and 2 patients with bladder cancer (2/90; 2.2%). Only 2 cases from the control group were HPV positive (both HPV-18; 2/30; 5%). The difference between the RTR and control groups was not significant (P > .05).

Conclusion

Malignant tumors in the urinary system following renal transplantation did not seems to be associated with infection by HPV-16 or -18.     

Detection of human papillomavirus DNA in cancer of the urinary bladder by in situ hybridisation.

The association of the human papillomavirus (HPV) with cancer of the urinary bladder was assessed by in situ hybridisation using probes selective for HPV types 6/11 and 16/18 DNA. No hybridisation signal was detected with the type 6/11 probe on 100 formalin-fixed, paraffin-embedded bladder tumours sampled. However, when the same samples were hybridised with the HPV type 16/18 DNA probe, 11 of 66 (16.6%) papillary and 1 of 10 (10%) solid transitional cell carcinomas gave positive signals. These results suggest the involvement of HPV in cancer of the bladder, although the frequency of multiple HPV types in these tumours is uncertain.     

Human papillomaviruses in urological malignancies: A critical assessment

ObjectivesInfection with human papillomaviruses (HPVs) is intimately associated with anogenital tract malignancies including cervical and vulvar cancer, a subset of oropharyngeal cancers and certain types of skin cancer. A number of urological tumors have likewise been suggested to be associated with high-risk HPV infection; however, many studies are hampered by a limited number of detection methods. The goal of this review article is to define a set of key criteria when implicating a virus in a human cancer and to apply these criteria to HPV infection in urological cancers.Materials and methodsWe performed a survey of the literature to corroborate the evidence to support a causal relationship between HPV infection and major urological malignancies.ResultsA number of previous reports have implicated HPVs in urological malignancies including penile, prostate, and bladder cancer. Most reports, however, rely only on a limited number of detection methods and frequently use contamination-prone polymerase chain reaction based methods. To firmly establish a link between a viral infection and a human malignancy, it is paramount that an array of techniques is employed and that the virus is ultimately traced by either direct visualization or, in the case of viral genome that has integrated into the host genome, detection of viral genes and gene products as well as functional cellular perturbations. In any case, seroepidemiological studies are likewise crucial to support the evidence. Such evidence for a role of HPV in urological malignancies based on currently available techniques is only present for penile squamous cell carcinomas.ConclusionsAn increasing number of immunocompromised patients as well as novel developments in patient care may change the spectrum of HPV-associated neoplasms. This is examplified by results demonstrating a role of HPVs in rare urothelial carcinomas with squamous differentiation in patients with neurogenic bladder. Hence, it is important to keep HPV infection in mind when confronted with unusual disease manifestations of the urogenital tract.