口咽鳞癌组织中人乳头状瘤病毒DNA状态与p16蛋白表达的相互关系及临床意义

目的 探讨联合检测口咽鳞癌组织标本中人乳头状瘤病毒(HPV)的感染情况与p16蛋白表达的临床意义.方法 选择66例口咽鳞癌活检或手术标本,采用“三明治”技术对组织标本进行切片,以SPF10-DNA LiPA分型方法检测HPV-DNA状态,采用免疫组织化学法检测p16蛋白的表达情况,分析不同HPV-DNA和p16蛋白表达患者的预后.结果 66例口咽鳞癌患者中,HPV-DNA阳性11例,阳性率为16.7％ (11/66).11例HPV-DNA阳性患者中,p16蛋白阳性9例,阴性2例.55例HPV-DNA阴性患者中,p16蛋白阳性12例,阴性43例.在口咽鳞癌中,HPV-DNA的表达与p16蛋白的表达具有显著的相关性(P ＜0.001).HPV-DNA和p16蛋白均为阳性组(A组)9例,HPV-DNA阳性/p16蛋白阴性或HPV-DNA阴性/p16蛋白阳性组(B组)14例,HPV-DNA和p16蛋白均为阴性组(C组)43例.A组、B组和C组患者的3年总生存率分别为100％、77.8％和42.0％,差异有统计学意义(P=0.001);3年疾病特异性生存率分别为100％、77.8％和46.4％,差异有统计学意义(P =0.004).结论 在口咽鳞癌中,HPV-DNA的表达与p16蛋白的表达具有显著的相关性,p16蛋白是较为可靠的预测HPV状态的替代标志.联合检测HPV-DNA和p16蛋白的状态,有助于更加准确地对口咽鳞癌进行分层,并判断预后.     

HSP70、p53和c—myc在宫颈鳞癌中的表达及与HPV16／18感染的关系

目的探讨热休克蛋白70(heat shock protein 70,HSP70)、抑癌基因p53、原癌基因c-myc在宫颈鳞癌中的表达及其与人乳头瘤病毒(HPV)感染的关系。方法采用免疫组化技术检测45例宫颈鳞癌、12例宫颈原位癌和20例正常宫颈组织中HSP70、p53、c-myc蛋白的表达,同时采用聚合酶链反应(PCR)技术检测HPV16/18的感染情况。结果①宫颈原位癌和宫颈癌中HSP70、p53、c-myc蛋白表达均显著高于正常宫颈组织。②HPV16/18阳性组HSP70表达明显高于HPV16/18阴性组,HPV16/18阴性组p53表达明显高于HPV16/18阳性组,但c-myc表达差异无显著性。结论在HPV感染的应激状态下,HSP70在宫颈鳞癌中过表达,p53则表达降低,c-myc蛋白的表达与HPV感染无相关性。     

p53Arg72Pro多态性与细胞周期调控蛋白在HPV相关子宫颈鳞癌中的表达及意义

目的探讨p53Arg型和p53Pro型蛋白功能失活与细胞周期调控异常在HPV相关子宫颈鳞癌发生机制中的作用。方法应用免疫组织化学方法检测p53、CyclinD1、Cdk4、Rb磷酸化及Ki-67蛋白表达（PI）。结果在HPV阳性p53蛋白阴性的子宫颈鳞癌组中CyclinD1、Cdk4蛋白表达、Rb蛋白磷酸化率及PI值均明显高于子宫颈对照组（χ2=16.878,P〈0.01;χ2=21.120,P〈0.01;χ2=40.36,P〈0.01;t=30.53,P〈0.01）。CyclinD1与Cdk4蛋白表达、Rb磷酸化率均呈正相关（P〈0.01）,Rb高磷酸化组中的PI值高于低磷酸化组（P〈0.05）。p53Pro型子宫颈鳞癌组中CyclinD1蛋白表达和PI值均高于p53Arg型组和p53Pro/Arg型组（P〈0.05）。结论 HPV阳性的子宫颈鳞癌组中部分p53蛋白表达阴性,提示该部分p53蛋白被E6蛋白降解。在p53阴性的子宫颈鳞癌组织中PI值增高与CyclinD1-Cdk4-Rb磷酸化途径有关。p53Pro型组比p53Arg型组蛋白可能具有较强的细胞周期阻滞功能。     

乳腺癌中HPV与p53、Rb蛋白表达的关系及对预后的影响

目的:研究乳腺癌中HPV与p53、Rb蛋白表达的关系及对预后的影响。方法:免疫组化法检测96例乳腺癌组织中p53、Rb蛋白的表达,采用流式荧光杂交技术检测标本中HPV的感染情况并分型。SPSS 17.0软件对试验结果进行统计分析。结果:96例乳腺癌标本中,p53、Rb、HPV的阳性率分别为59.38%、87.50%、7.29%,p53阳性表达率与淋巴结转移率呈正相关（P<0.05）,p53、Rb表达与HPV感染均无明显相关性（P=0.26,P=0.59）。p53、Rb及HPV的表达与预后无相关性（P=0.628,P=0.084,P=0.699）。结论:HPV与p53、Rb的表达在乳腺癌中无明显相关性,p53高表达的患者淋巴结转移率较高。     

外阴鳞癌组织中p14ARF表达与人乳头状瘤病毒感染状态关系的研究

目的:探讨p14ARF表达与不同HPV感染状态的外阴鳞癌发生和发展的关系.方法:采用RT-PCR和蛋白印迹技术检测42例外阴鳞癌组织、10例正常外阴组织中p14ARF基因mRNA及蛋白表达水平,PCR技术检测其HPV-DNA,分析HPV阳性组和阴性组外阴鳞癌组织中,p14ARF基因mRNA及蛋白表达与其临床病理特性的关系.结果:外阴鳞癌组p14ARF基因mRNA及蛋白表达强度为0.92±0.24和1.09±0.11;正常外阴组分别为0.93±0.06和1.02±0.38,两组比较差异无统计学意义,P>0.05;低、中和高分化鳞癌组p14ARF基因mRNA表达分别为0.98±0.11、0.94±0.15和0.91±0.07;p14ARF基因蛋白表达低、中和高分化分别为1.14±0.24、1.11±0.27和1.07±0.29,3组间比较差异均无统计学意义,P>0.05.p14ARF基因mRNA及蛋白在HPV阳性组表达强度分别为0.99±0.04和1.11±0.02,HPV阴性组表达强度分别为0.61±0.07和0.76±0.06, 两组比较差异有统计学意义,P<0.05.结论:p14ARF可能参与了外阴鳞癌的发生发展过程,并可能成为HPV感染的外阴鳞癌发生进展相关的基因.     

乳腺浸润性导管癌组织中 HPV18 DNA 及 p53 蛋白的表达

目的：探讨人乳腺浸润性导管癌组织中HPV18DNA和p53蛋白的表达及二者间的关系。方法：采用分子原位杂交和免疫组化技术检测60例乳腺浸润性导管癌、30例乳腺腺病和30例正常乳腺组织中HPV18DNA和p53蛋白。结果：癌组中HPV18DNA阳性和HPV18DNA阴性两组间p53蛋白的阳性表达均有显著性差异（P〈0．03），HPV18DNA阳性组与p53蛋白阳性表达呈显著负相关（r=-0．2954、P〈0．04）。结论：HPV18感染后导致野生型p53的灭活和降解可能涉及HPV18感染后人乳腺上皮细胞癌变的转化过程。     

口咽鳞癌组织中HPV16感染和p16蛋白表达的临床意义

目的:检测口咽鳞癌中HPV16感染及p16表达率,并分析临床意义。方法:采用原位杂交法检测60例口咽鳞癌组织中HPV16的表达,免疫组化法检测95例口咽鳞癌组织p16的表达情况,分析HPV16与p16的相关性,总结上述两种蛋白在口咽癌中的临床意义。结果:口咽鳞癌组织中HPV16感染率为48.3%（29/60）,p16阳性表达率为52.6%（50/95）,60例标本中HPV16感染与p16表达显著相关（P<0.001）,用p16表达预测HPV16感染的正确率可达90%（54/60）,跟国内外报道一致。p16阳性组相比p16阴性组,吸烟、饮酒、非嚼槟榔、T3+T4、有淋巴结转移、III-IV临床分期和高分化者所占比例更低,但不具有统计学差异(P>0.05)。平均随访时间38个月,随访率65.3%（62/95）,62例口咽鳞癌病人中p16阳性组和p16阴性组的3年总体生存率和无瘤生存率分别为70.4%对比40.0%（P=0.008）和63.0%对比25.7%（P=0.004）。总体生存率多因素分析提示,p16阴性病人死亡风险是p16阳性病人的2.12倍(HR 0.471,95%CI 0.201-1.103,P=0.083),无瘤生存率多因素分析提示,p16阴性病人死亡风险是p16阳性病人的2.36倍(HR 0.432,95%CI 0.201-0.891,P=0.024),p16与口咽癌的预后密切相关。结论:p16表达和HPV16表达存在紧密相关性,p16可以替代HPV16来预测预后,p16蛋白阳性口咽癌患者具有非吸烟、非饮酒、嚼槟榔和肿瘤分化程度低的趋势,检测p16表达对口咽癌病人预后判断有重要的提示意义。     

HPV感染及p53与Cdc2蛋白表达对喉癌术后复发的预测价值

目的检测人乳头状瘤病毒(human papilloma virus,HPV)及p53和Cdc2蛋白表达与喉鳞状细胞癌发生及复发的关系。方法选取2003-01-01-2011-12-31唐山市协和医院92例喉鳞状细胞癌手术切除标本,采用PCR-反向点杂交方法检测喉癌组织中HPV感染情况,SP免疫组化方法检测p53和Cdc2蛋白表达,收集临床病理资料并进行随访。结果随访>2年的喉癌患者中有18例出现复发(复发组),74例无复发(未复发组)。92例喉癌组织中HPV的感染率为19.57%(18/92);喉癌组织中p53和Cdc2蛋白的阳性表达率分别为60.87%(56/92)和65.22%(60/92)。HPV感染、p53及Cdc2蛋白表达与喉癌的分化程度、分期均无明显相关性,P值均>0.05;HPV阳性和阴性喉癌患者术后复发率分别为0(0/18)和24.32%(18/74),差异有统计学意义,χ2=4.007,P=0.045;喉癌复发组和未复发组中的p53蛋白表达率分别为72.22%(13/18)和58.11%(43/74),差异无统计学意义,χ2=0.691,P=0.406;喉癌复发组和未复发组中Cdc2蛋白表达率分别为88.89%(16/18)和59.46%(44/74),差异有统计学意义,χ2=4.307,P=0.038。喉癌组织中HPV的感染与p53(χ2=0.316,P=0.574)及Cdc2(χ2=0.574,P=0.487)蛋白表达均无相关性。结论喉癌组织中存在着一定程度的HPV感染及p53、Cdc2蛋白质的表达,但三者与喉癌的分期及分化程度无关;HPV阳性患者术后复发率低于HPV阴性患者;Cdc2蛋白表达阳性者术后复发率高于阴性患者。HPV感染、p53及Cdc2 3种因素可能参与了喉癌的发生、发展及复发过程。     

原发性肺癌中高危型HPV感染及p53蛋白表达的相关性研究

目的:通过检测高危型人乳头瘤病毒(human papillomavirus,HPV)与p53蛋白在原发性肺癌中的表达情况,探讨高危型HPV感染与p53蛋白表达的相关性及其在原发性肺癌发生和发展中可能的作用。方法:收集2014年4月—2015年6月在新疆医科大学附属肿瘤医院行手术切除的51例原发性肺癌和26例肺良性病变患者的资料,采用第二代杂交捕获法(hybrid capture 2,HC2)检测原发性肺癌和肺良性病变支气管上皮细胞中高危型HPV的感染情况,免疫组织化学法检测原发性肺癌组织和肺良性病变组织中p53蛋白的表达情况。结果:原发性肺癌患者支气管上皮细胞中高危型HPV感染检出率为15.69%,明显高于肺良性病变患者支气管上皮细胞中的0%,差异有统计学意义(P<0.05);原发性肺癌组织p53蛋白的阳性表达率为52.94%,显著高于肺良性病变组织的11.54%,差异有统计学意义(P<0.001)。HPV阳性组中p53蛋白的阳性表达率为75.00%,高于HPV阴性组的48.84%,差异有统计学意义(P<0.05)。结论:HPV感染可能是原发性肺癌发生的危险因素之一,其可能通过使p53失活而促进肺癌的发生和发展。     

乳腺浸润性导管癌组织中HPV16DNA与p53蛋白表达的研究

 目的 检测人乳腺浸润性导管癌组织中HPV16DNA和p53蛋白，探讨HPV16型感染和p53蛋白表达与人乳腺癌之间的关系。方法 采用分子原位杂交和免疫组化技术检测和分析50例乳腺浸润性导管癌、30例正常乳腺组织中HPV16DNA和p53蛋白二者之间的表达关系。结果 癌组中HPV16DNA阳性和HPV16DNA阴性两组阃p53蛋白的表达均有显著性差异（P〈0．01），HPV16DNA阳性组显示与p53蛋白表达呈负相关（P〈0．02）。结论 HPV16感染后导致野生型p53的降解可能涉及HPV16感染后人乳腺上皮细胞癌变的病理发生过程。     

肺鳞癌与人乳头瘤病毒相关性研究

目的:了解肺鳞癌组织中人乳头瘤病毒存在情况,并分析人乳头瘤病毒与肺癌发生部位、性别因素、角化程度、吸烟因素的关系.方法:采用地高辛标记的DNA探针对肺癌组织进行原位杂交检测,同时结合临床资料分析多种因素的相关性.结果:肺鳞癌组织,鳞状上皮生化组织、粘膜慢性炎组织HPV检出率分别为48.4%,27.4%,5.6%,各组比较差异显著(P＜0.01),中心型肺癌与周围型肺癌组织HPV检出率分别为50.6%和4.4%,两组比较差异显著(P＜0.01).男女两组HPV检出率分别为46.1%和54.3%,差异无显著性(P＞0.05).吸烟组HPV检出率为50.6%,非吸烟组为43.9%,差异不显著(P＞0.05).结论:肺癌组织中存在一定比例HPV感染,HPV感染与肺鳞癌关系密切,中心型肺癌与HPV感染密切相关,HPV感染与性别及吸烟因素未发现明显相关性.人乳头瘤病毒感染可能与肺癌发生发展有关.     

Human papillomavirus type 16 and 18 infection is associated with lung cancer patients from the central part of China

Infection with specific high-risk human papillomavirus (HPV) types 16 and 18 have been strongly associated with the genesis of various neoplasms in humans, though such study in lung cancer is limited and the results are controversial. In the present study, we collected and explored 313 fresh lung tumor specimens for the presence of HPV with polymerase chain reaction and non-isotopic in situ hybridization. We found that 44.1% of (138/313) non-small cell lung carcinoma (NSCLC) samples were positive for HPV detection, while 4.2% (4/96) of lung benign controls were positive for HPV 16 and 18 DNA. HPV infection was significant between lung squamous cell carcinoma and adenocarcinoma as well as smoking and non-smoking patients. In HPV-positive lung cancer tissues, abnormal p53 protein accumulation was seen in 97 of the 138 carcinomas (70.3%) and expression of pRb in 54 of the 138 carcinomas (39.1%). There was an obvious relationship between the presence of papilloma viral DNA and abnormal p53 protein accumulation and pRb depletion. Cell proliferation and apoptosis were correlated with HPV infection in NSCLC samples. Our data confirm the high prevalence of HPV in lung carcinomas in the central part of China and suggest the possible mechanism of the carcinogenic role of HPV in these carcinomas.     

Human papilloma virus and p53 expression in bladder cancer in Egypt: Relationship to schistosomiasis and clinicopathologic factors

The aim of the current study was to compare the role of p53 and human papillomavirus (HPV) in schistosomiasis-related and schistosomiasis-unrelated carcinoma of the urinary bladder. To achieve this aim, we investigated 114 bladder carcinomas for p53 oncoprotein expression by immunohistochemistry and for human papillomavirus by in situ hybridization technique. The results revealed that 64 tumors (56.1%) were schistosomiasis-associated. Sixty seven (58.8%) were transitional cell carcinomas and 32 (28%) were squamous cell carcinomas. The remaining 15 tumors (13.2%) included adenocarcinomas and sarcomatoid carcinomas. In both schistosomiasis-associated and non-associated carcinomas, p53 oncoprotein expression was significantly higher in poorly differentiated tumors. However, it was significantly higher in locally more invasive tumors in the schistosomal carcinomas only. HPV types 16/18 could be detected in 1 of the 114 bladder carcinomas (0.95%), which was schistosomiasis-related squamous cell carcinoma in situ. These results suggest that p53 immunohistochemistry can be a prognostic factor in both schistosomal and nonschistosomal bladder cancer. More importantly, HPV does not seem to play a role in the pathogenesis of either type of bladder cancer in our country.(Pathology Oncology Research Vol 12, No 3, 173–178) An erratum to this article is available at .     

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。     

Is there an epidemiological link between human papillomavirus DNA and basaloid squamous cell carcinoma of the pharynx?

The epidemiological link between human papillomavirus (HPV) DNA and basaloid squamous cell carcinoma (BSCC) of the pharynx was studied. The expression of p53 protein was evaluated by immunohistochemical analysis. The presence of HPV DNA was evaluated by polymerase chain reaction amplification and "in situ" hybridization. The tobacco and alcohol consumption and the clinical outcomes of nine patients with BSCC of the pharynx are compared with site and stage matched 109 conventional squamous cell carcinoma (SCC) patients. The BSCC specimens were analyzed for the presence of HPV DNA and p53 expression. We did not find any significant differences in tobacco and alcohol consumption between patients with BSCC and patients with SCC. No HPV DNA was detected in BSCC, and p53 overexpression was found in five (55%) of the cases. Our results do not support an etiological link between HPV DNA and BSCC.     

A subset of head and neck squamous cell carcinomas exhibits integration of HPV 16/18 DNA and overexpression of p16INK4A and p53 in the absence of mutations in p53 exons 5-8

Besides well-known risk factors such as tobacco use and alcohol consumption, oncogenic human papillomavirus (HPV) infection also has recently been suggested to promote head and neck tumorigenesis. HPV is known to cause cancer by inactivation of cell cycle regulators p53 and pRb via expression of viral oncoproteins E6 and E7. This indicates that p53 mutations are not a prerequisite in HPV-induced tumor development. However, discrepancy exists with respect to the frequency of head and neck squamous cell carcinomas (HNSCC) harboring DNA of oncogenic HPV and the fraction of these tumors showing p53 mutations. In our study, we examined the frequency of HNSCC demonstrating HPV 16/18 integration as identified by fluorescence in situ hybridization (FISH) and investigated their p53 (mutation) status by immunohistochemistry and single-strand conformation polymorphism (SSCP) analysis of exons 5-8. Paraffin-embedded, archival biopsy material from 27 premalignant mucosal lesions and 47 cases of HNSCC were analyzed. Ten of the 47 (21%) HNSCC unequivocally exhibited HPV 16 integration, including 8 of 12 (67%) tonsillar carcinomas. This is supported by the immunohistochemical detection of p16(INK4A) overexpression in all 10 HPV-positive tumors. Although FISH is considered to be less sensitive than PCR-based methods for HPV detection, our data clearly demonstrate clonal association of HPV with these tumors, as illustrated by the presence of integrated HPV 16 in both the primary tumor and their metastases in 2 patients. In contrast, HPV 16/18 DNA could not be detected in the premalignant lesions. In 30 of 47 (64%), HNSCC accumulation of p53 was observed, including 8 of the 10 HPV-positive carcinomas. However, in none of the latter cases could mutations in exons 5-8 be identified, except for a polymorphism in codon 213 of exon 6 in one patient. Evaluation of clinical data revealed a significant inverse relation between tobacco use with or without alcohol consumption, and HPV positivity of the tumors.     

Papillomavirus infection of the lower genital tract: detection of viral DNA in gynecological swabs

A total of 311 smears from the lower genital tract were examined by the filter in situ hybridization method to identify human papillomavirus (HPV) DNA. Of these 311 smears, 229 came from clinically and cytologically negative patients and served as a control group. In this group HPV-DNA was detected in 5 cases (2.2%). Of 82 cytologically positive cases (25 confirmed by histology) 56 (68%) contained HPV-DNA. A high prevalence of HPV 6/11 and absence of HPV 16/18 was found in cases with cytological signs of permissive HPV infection. In mild and moderate dysplasia all viruses occurred at almost the same frequency. In severe dysplasia/carcinoma in situ HPV 16/18 was found 5 times more frequently than HPV 6/11. HPV 16/18 was identified in all 4 invasive cancer cases. Cervical irrigation of colposcopically suspect areas was performed in 15 cytologically and HPV-DNA positive cases using the hydrodynamic filtration method. In 12 cases only the cells obtained from the colposcopically positive areas contained HPV-DNA. The sensitivity and reproducibility of the filter in situ hybridization was shown by: comparing the results obtained by HPV-DNA hybridization using Southern blot analysis of tumor biopsies; analysing the correlation of cytologic diagnosis and presence of HPV-DNA in follow-up examinations, and diagnosing presence or absence of HPV-DNA in parallel filters from the same patients.