HLA-DRB1*1501和DQB1*0602与新疆哈萨克族HPV 感染及食管癌发生的相关性研究*

目的：从基因水平探讨新疆哈萨克族食管鳞癌的HPV 16E6 感染及食管鳞癌发生与HLA-DRB1,DQB 1 等位基因的遗传易感性,为寻找哈萨克族食管鳞癌的易感基因提供参考。方法：采用聚合酶链反应（PCR ）技术检测200 例哈萨克族食管鳞癌和150 例哈族萨克正常人群HPV 16E6 基因的表达情况,运用序列特异性引物聚合酶链反应技术（PCR-SSP ）,检测新疆哈萨克族食管鳞癌患者200 例,哈萨克族正常人群食管膜膜150 例的HLA-DRB1*1501和HLA-DQB1*0602的分布。结果：新疆哈萨克族食管鳞癌患者HPV 16E6 感染率为41% ,明显高于哈萨克族正常人群感染率的14%（P<0.001,OR= 3.94）;HPV 16E6 感染与哈萨克族正常人群HLA-DRB1*1501,HLA-DQB1*0602的无相关性（P>0.05）;新疆哈萨克族食管鳞癌患者HLA-DRB1*1501和HLA-DQB1*0602基因分布频率显著高于哈萨克族正常人群（0.455：0.232,P<0.001,OR= 2.78;0.69比0.554,P=0.006,OR= 1.80）;HLA-DQB1*0602基因阳性率在中低分化鳞癌组中（68.8%）的分布高于高分化鳞癌组（31.2%）,差异有统计学意义（P<0.05）。 结论：HPV 16E6 的感染可能是新疆哈萨克族食管癌发生的重要因素之一。HLA-DRB1*1501和HLA-DQB1*0602是哈萨克族食管鳞癌的易感基因,其中HLA-DQB1*0602与哈萨克族食管鳞癌的分化程度有关。     

HPV16感染与新疆哈萨克族食管癌发病及临床病理学研究

背景与目的:人乳头状瘤病毒(human papilloma virus,HPV)作为食管癌发生重要的环境因素备受许多学者的关注,但其相关性未得到一致性公认,尤其对于高发的新疆哈萨克族食管癌.本研究探讨HPV16感染与新疆哈萨克族食管痛发生、发展的相关性.方法:采用半巢式聚合酶链反应(polymerase chain reaction,PCR)技术检测100例新疆哈萨克族食管癌患者癌组织和100例新疆哈萨克族正常人食管正常黏膜组织HPV16 E6的感染情况.结果:新疆哈萨克族食管癌患者HPV16 E6感染率为46%,明显高于新疆哈萨克族正常人群感染率的22%(P<0.001,OR=3.020);HPV16 E6感染与新疆哈萨克族食管癌患者的发病年龄、性别、肿瘤生长部位、淋巴转移、分化程度无明显相关性(P均>0.05).结论:HPV16 E6感染与新疆哈萨克族食管癌发病密切相关,是其发病的重要因素之一.     

新疆哈萨克族食管鳞癌中p16蛋白的表达及其意义

目的研究p16蛋白表达与新疆哈萨克族食管鳞癌生物学行为的关系。方法应用免疫组织化学SP法检测30例食管鳞癌和60例正常食管组织中p16蛋白的表达。结果60例正常组织中p16蛋白阳性表达率为88.33%(53/60);30例食管鳞癌组织中p16蛋白表达阳性率为46.67%（14/30）。随着恶性程度增加及病程进展,p16蛋白阳性率逐渐下降,p16在高分化鳞癌组阳性表达率(75%)显著高于低分化鳞癌组(25%)(P<0.05)。结论p16蛋白表达缺失可能与哈萨克族食管癌的发生发展有关。     

高危型人乳头瘤状病毒感染与湖北十堰食管鳞癌发生的关系研究

目的:检测高危型人乳头瘤病毒(human papillomavirus,HPV)在十堰地区食管鳞癌(esophageal squa-mous cell carcinoma,ESCC)组织中的感染率,分析其相关性.方法:收集十堰地区87例食管鳞癌组织和50例对照组食管组织,实时荧光定量PCR法检测组织中HPV16感染率.结果:食管鳞癌组织和对照组食管组织中HPV16的检出率分别为57.5%和36.0%,食管鳞癌组织中HPV16感染率显著高于对照组(P=0.016<0.05).分析食管鳞癌患者年龄、性别、病理分级、大体分型、浸润深度、淋巴结侵犯以及TNM分期与HPV16感染的关系,差异均无统计学意义.HPV16感染与食管鳞癌患者临床特征未见显著相关性.结论:HPV16感染可能是十堰地区食管鳞癌发生的因素之一,针对高危型HPV感染的防治对于该地区食管鳞癌的发生具有重要意义.     

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.     

172例新疆哈萨克族食管鳞状细胞癌临床病理资料分析

目的:探讨新疆哈萨克族食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)流行病学特征,并结合临床资料及免疫表型分析.方法:收集了172例新疆哈萨克族ESCC患者资料,分析其临床病理资料,对标本进行免疫组化染色观察.结果:69.2％新疆哈萨克族ESCC患者＜65岁,病变多在食管的中下段.其病理学分期,多以T3 +T4为主.哈萨克族ESCC组织CyclinD1阳性表达显著高于汉族ESCC(P=0.025).结论:新疆哈萨克族ESCC具有种族特异性,推测CyclinD1是哈萨克族食管癌的主要致病基因.     

人乳头状瘤病毒感染与食管癌发生关系的研究

为了探索人乳头状瘤病毒(HPV)与食管癌发生的关系,对食管鳞癌、癌旁鳞状上皮及乳头状瘤进行组织病理学研究,并用生物素标记的HPV_(16)DNA探针进行原位杂交。结果:癌旁鳞状上皮有凹空细胞等HPV感染的组织形态学改变者为65.2%(15/23),HPV_(160)DNA检出率为56.5%(13/23);食管鳞癌有凹空样癌细胞者为27.8%(10/36),HPV_(16)DNA检出率为50.0%(18/36);6例乳头状瘤均检出HPV_(16)DNA(100.0%)。不同分化程度的食管鳞癌(Ⅰ、Ⅱ、Ⅲ级)HPV_(16)DNA检出率分别为100.0%(6/6),60.0%(9/15)和20.0%(3/15)。研究结果提示,HPV感染与食管鳞癌发生有关,并与其分化和某些组织学特征有关。     

新疆哈萨克族食管癌组织中HPV16 E6、E7基因的检测与p53的关系

目的探讨人乳头状瘤病毒(HPV)16型感染在新疆哈萨克族(哈族)食管癌发病中的作用,并分析HPV16感染与p53过表达之间的关系.方法采用聚合酶链(PCR)技术,检测41例食管鳞状细胞癌组织中HPV16 E6与E7基因表达差异;用LSAB免疫组织化学方法分析p53蛋白的表达.结果癌组织中HPV16 E6与E7基因阳性检出率分别为34.15%(14/41)和63.41%(26/41);用免疫组化检出p53蛋白阳性率分别在HPV16 E6阳性组(57.1%)与HPV16 E6阴性组(14.8%)间、HPV16 E7阳性组(42.3%)与HPV16 E7阴性组(6.7%)间均存在显著性差异(P＜0.05);用PCR检出HPV16 E6、E7基因在食管癌的高分化组、中低分化组中的检出率分别为7.69%(1/13)、46.43%(13/28)和38.46%(5/13)、75.00%(21/28),差别均有统计学意义(P＜0.05).结论提示p53基因突变与HPV16感染在哈族食管癌的发病过程中存在相互促进作用;另外,HPV16 E6与E7基因和哈萨克族食管癌病理组织学分级的生物学行为密切相关.     

河南地区食管鳞癌和贲门腺癌与人乳头瘤病毒感染相关性的分析

目的:了解人乳头瘤病毒16型(HPV16)与河南食管癌高发地区食管鳞癌、贲门腺癌发生的关系.方法:利用聚合酶链式反应(PCR)对高发区食管鳞癌组织(44例)、贲门腺癌组织(18例)进行HPV-DNA检测.结果:食管鳞癌及贲门腺癌组织中均检测到HPV16 E6 DNA表达,但食管鳞癌HPV16 E6 DNA表达(84%,37/44)明显高于贲门腺癌(44%,8/18),P<0.01.食管鳞癌及贲门腺癌组织中,高危型HPV16 E6 DNA表达与患者年龄、性别、分化程度、浸润程度,淋巴结转移以及肿瘤分级无相关性,P值均＞0.05.结论:同一地区食管鳞癌和贲门腺癌均有不同程度HPV感染,提示HPV可能是两者共同相关致病危险因素,高危型HPV16感染可能在食管鳞癌和贲门腺癌发生中起重要作用.     

人类乳头状瘤病毒与肺癌的病因关系的研究

目的　研究人类乳头状瘤病毒 (HPV)与肺鳞癌发生的关系。方法　采用聚合酶链反应(PCR)技术检测石蜡包埋肺鳞癌组织标本 50份、肿瘤组织旁鳞状化生上皮标本 3 0份、正常支气管粘膜3 0份中的HPVDNA。结果　三种标本中 ,HPVDNA的检出率分别为 2 6%、3 6.7%和 1 0 % ,其中以HPV1 6型所占比例最高。鳞状细胞癌和鳞状化生上皮标本中 ,HPV1 6与HPV6/ 1 1的检出率无明显差异 (P >0 .0 5)。结论　结果表明HPV感染在肺鳞癌的发生中可能起了重要的作用。鳞状上皮化生似可视为癌前病变。PCR技术较体外杂交技术和DNA探针有更高的敏感性。     

Detection of Human Papillomaviral Infection on Kazakh Patients with Esophageal Squamous Cell Carcinoma in Xinjiang

OBJECTIVE To investigate the detection rate of human papilloma virus (HPV) DNA in the Kazakh esophageal carcinoma (EC) patients of Xinjiang.METHODS We detected the prevalence of a HPV gene in tumor tissues from 318 esophageal squamous cell carcinoma (ESCC).Tumor tissues were kept in formalin and embedded in paraffin.One hundred seventeen samples used crude cell suspension, while the other 201 used the method of DNA extraction with phenol-Tris/chloroform. We analyzed the relevance to EC of Kazakh's in Xinjiang.RESULTS In the ESCC samples of Kazakh's in Xinjiang, total detection rate for HPV DNA was 64.5% (205/318). The positive rate of HPV in group of crude cell suspensions was 82.9% (97/117) compared with the rate of 53.7% (108/201) in the group of DNA extraction. The results in the two groups showed significant diffference (X2 = 5.711, P < 0.05).CONCLUSION HPV DNA infection may be one of the most important factors related to EC of Kazakh's in Xinjiang.     

Evaluation of human papilloma virus infection in patients with esophageal squamous cell carcinoma from the Caspian Sea area, north of Iran

Introduction: HPV has been found repeatedly in esophageal squamous cell carcinoma (ESCC) tissues. However,reported detection rates of HPV DNA in these tumors have varied markedly. Differences in detection methods,sample types, and geographic regions of sample origin have been suggested as potential causes of variation. Wehave reported that infection of HPV DNA in ESCC tumors depends on anatomical sites of esophagus of thepatients from Mazandaran, north of Iran. Materials and Methods: HPV DNA was examined in 46 upper, 69middle and 62 lower third anatomical sites of esophageal squamous cell carcinoma specimens collected fromMazandaran province in north Iran, near the Caspian Littoral as a region with high incidence of ESCC. HPVL1 DNA was detected using Qualitative Real time PCR and MY09/MY11 primers. Results: 28.3% of upper,29% of middle and 25.8% of lower third of ESCC samples were positive for HPV DNA. 13.6% for males and14.1% for females were HPV positive in all samples. Conclusions: HPV infection is about one third of ESCCin this area. Findings in this study increase the possibility that HPV is involved in esophageal carcinogenesis.Further investigation with a larger sample size is necessary.     

Human papillomavirus in high- and low-risk areas of oesophageal squamous cell carcinoma in China

To examine the potential roles of human papillomavirus (HPV) in oesophageal squamous cell carcinoma (ESCC) development, we examined the presence of HPV DNA in paraffin-embedded ESCC tissues collected from two areas with different ESCC incidence rates in China, that is, Gansu (n=26) and Shandong (n=33), using PCR with SPF10 primers, or PCR with GP5+/GP6+ primers combined with Southern blot hybridisation. HPV genotype was determined by the INNO-LiPA HPV genotyping kit. HPV DNA was detected in 17 cases (65%) in Gansu, where ESCC incidence is much higher than in Shandong, where HPV was positive in two samples (6%). HPV genotypes 16 and 18 were detected in 79 and 16% of HPV-positive samples, respectively. Real-time PCR analysis suggested the presence of integrated form of HPV DNA in all the HPV-16-positive samples, but its viral load was estimated to be only <1-2 copies cell(-1). We could not detect HPV 16/18 E6 protein expression by immunostaining in any of the HPV-16-positive samples. Neither p16(INK4a) nor p53 expression was related to HPV presence in ESCCs. Further studies seem warranted to examine the possible aetiological roles of HPV in ESCC.     

Viral load of HPV in esophageal squamous cell carcinoma

We previously reported the presence of HPV DNA in esophageal squamous cell carcinoma (ESCC) cases from Hong Kong and Sichuan. The role of HPV in the carcinogenesis of ESCC remains unclear, partly due to the large variations in infection rates reported by different studies. While some of these variations may truly reflect different HPV infection rates in ESCC among different geographic regions, differences in sensitivity and specificity of the detection methods used also contribute. In the present study, we used quantitative real-time PCR to determine the copy numbers of HPV-16 and HPV-18 in ESCC from 5 different regions of China with different incidence rates of ESCC. Conforming to our previous reports, HPV infection was detected in 2-22.2% of samples. Infection with HPV-16 was again shown to be more common than that with HPV-18 among Chinese ESCC patients. The copy number of HPV-16 in these ESCC cases ranged from < or =1 to 157 copies/genome equivalent, with 65% of samples harboring fewer than 10 copies/genome equivalent. The median copy number of HPV-18 was 4.9/genome equivalent. Assays were validated using cervical carcinoma cell lines with known copy numbers of HPV-16 or HPV-18. The relatively low HPV copy number and infection rate in ESCC suggest that HPV is unlikely to play as essential a role in the carcinogenesis of ESCC as in cervical cancer. However, with the consistent detection of oncogenic HPVs in ESCC from some regions of China, the possibility of HPV infection being one of the multiple risk factors of ESCC in some geographic areas cannot be excluded.     

A quantitative polymerase chain reaction-enzyme immunoassay for accurate measurements of human papillomavirus type 16 DNA levels in cervical scrapings.

A quantitative polymerase chain reaction-enzyme immunoassay (Q-PCR-EIA) was developed to measure the amount of human papillomavirus (HPV) 16 DNA per genome equivalent in cervical scrapings. The quantitative approach was based on a combined competitive PCR for both HPV 16, using the general primer GP5+/6+ PCR, and beta-globin DNA. The two competitive PCRs involve co-amplification of target sequences and exogenously added DNA constructs carrying a rearranged 30 bp sequence in the probe-binding region. The accuracy of quantification by combining the two competitive PCR assays was validated on mixtures of HPV 16 containing cervical cancer cells of CaSki and SiHa cell lines. Comparison of this fully quantitative PCR assay with two semi-quantitative HPV PCR assays on a series of crude cell suspensions from HPV 16 containing cervical scrapings revealed remarkable differences in the calculated relative HPV load between samples. We found evidence that correction for both intertube variations in PCR efficiency and number of input cells/integrity of DNA significantly influence the outcome of studies on viral DNA load in crude cell suspensions of cervical scrapings. Therefore, accurate measurements on viral DNA load in cervical scrapings require corrections for these phenomena, which can be achieved by application of this fully quantitative approach.     

Detection of Merkel Cell Polyomavirus and Human Papillomavirus in Esophageal Squamous Cell Carcinomas and Non-Cancerous Esophageal Samples in Northern Iran

Human papillomavirus (HPV) infection is one of the hypothesized causes of esophageal squamous cell carcinoma (ESCC), but the etiological association remains uncertain. It was postulated that other infectious agents together with HPV may increase the risk of ESCC. The current investigation aimed to explore the presence of a new human tumor virus, Merkel cell polyomavirus (MCPyV), together with HPV in ESCC tumors and non-cancerous esophageal samples in northern Iran. In total, 96 esophageal samples (51 with ESCC, and 45 without esophageal malignancy) were examined. HPV DNA was detected in esophageal specimens of 16 out of the 51 ESCC cases (31.4 %) and 20 out of the 45 non-cancerous samples (44.4 %). Untypable HPV genotypes were recognized in high rates in cancerous (75.0 %) and non-cancerous (55.0 %) esophageal specimens. MCPyV DNA was detected in esophageal specimens of 23 out of the 51 ESCC cases (45.1 %) and 16 out of the 45 non-cancerous samples (35.6 %). The mean MCPyV DNA copy number was 1.0 × 10^?5 ± 2.4 × 10^?5 and 6.0 × 10^?6 ± 1.3 × 10^?5 per cell in ESCC cases and non-cancerous samples, respectively. There was no statistically significant difference between cancerous and non-cancerous samples regarding mean MCPyV DNA load (P = 0.353). A bayesian logistic regression model adjusted to the location of esophageal specimen and MCPyV infection, revealed a significant association between HPV and odds of ESCC (OR, 2.45; 95 % CI: 1.01–6.16). This study provides the evidence of the detection of the MCPyV DNA at a low viral copy number in cancerous and non- cancerous esophageal samples.     

Esophageal squamous cell cancer in patients with head and neck cancer: Prevalence of human papillomavirus DNA sequences

An etiologic role for human papillomavirus (HPV) infections in either head and neck (HNC) or esophageal carcinogenesis remains debatable. Patients with head and neck cancer are at high risk for developing a second esophageal squamous cell cancer (ESCC). The aim of our study was to determine whether HPV infections play a role in this multifocal carcinogenesis. Samples from 2 groups of HNC patients were studied: Random esophageal biopsies were collected from the first group of 60 patients who had been screened for asymptomatic ESCC. The second group consisted of 21 patients with pairs of HNC and ESCC. Both the fresh frozen biopsy samples of the first group and the paraffin-embedded specimens of the second group were evaluated for the presence of HPV DNA sequences by PCR amplification, cloning and sequencing. HPV DNA sequences were detected in 66.7% of normal/inflammatory (34/51) and dysplastic and malignant (6/9) esophageal tissues from HNC patients being screened endoscopically. Similarly, in the second group of 21 patients with both HNC and ESCC, HPV DNA sequences were demonstrated in 13 (61.9%) of the HNC biopsies and in 14 (66.7%) of the ESCC biopsies. The prevalence of high-risk-type HPV 16 was low (5/51, 9.8%) in normal/inflammatory esophageal mucosa but higher (10/24, 47.6%) in ESCC. The low-risk HPV 11 was present in 37.3% (19/51) of normal/inflammatory, 66.7% (4/6) of dysplastic and 28.9% (13/45) of the carcinoma samples. The same HPV type was present in only 3/21 pairs of HNC and ESCC samples, suggesting that a clonal expansion from the HNC to a subsequent ESCC, or visa versa, is unlikely. The high prevalence of "low-risk" HPV infections points to the need for studies on possible interactions of these infections with the use of alcohol and tobacco in the pathogenesis of these tumors.     

High-Risk and Low-Risk Human Papillomavirus in Esophageal Squamous Cell Carcinoma at Mazandaran, Northern Iran

Cancers are the second most common cause of non-accidental deaths in Iran, following cardiovascular deaths. Mazandaran, near the Caspian Littoral at north of Iran have identified as a several-high incidence area for Esophageal Squamous Cell Carcinoma (ESCC) in the world. Several associated risk factors, such as dietary and cultural habits, infectious agents, nutritional deficiencies, too much use of tobacco and alcohol and infection to certain DNA tumor viruses (HPVs), including environmental and genetic factors are attributed to this disease. To explore this issue, we analyzed HPV DNA prevalence and HPV types together in relation to tumor sites a high-incidence population. Archived tissue blocks from 46, 69 and 62 upper, middle and lower third of esophagus, respectively from ESCC patients were evaluated for the presence of HPV DNA by PCR using the degenerate HPV L1 consensus primer pairs MY09/MY11. The positive specimens were evaluated by Real-time PCR to determine HPV genotypes. From the 49 HPV positive cases, of ESCC patients, 5 (23.1 %), 11 (55 %) and 9 (56.3 %) of upper, middle and lower third of ESCC specimens, respectively were positive by at least one high and one low-risk HPV genotypes. In general, HPV45 and HPV11 were the most common high- risk and low-risk HPV genotypes in HPV L1 positive cases, respectively, followed by HPV6, HPV52 and HPV39. Therefore, the high prevalence of HPV DNA in different anatomical sites of ESCC patients from the Mazandaran region in North of Iran provides more evidence for a role of HPV in this cancer.     

Detection and identification of human papillomavirus in cervical adenocarcinoma

Objective:To evaluate the rate and types of human papillomavirus(HPV)infection in cervical adenocarcinoma.Methods:We detected and identified HPV in 67 lesions using PCR based reverse line blot hybridization and DNA sequencing.Among the 67 patients,53 were diagnosed as cervical adenocarcinoma and 14 as cervical adenosquamous carcinoma.First a fragment of 150 bp was amplified from the L1 region of HPV with GP5/GP6 primers.If the result was negative,a short fragment of 65 bp was amplified from the L1 region with SPFI/SPF2 primers.Results:6 cases were eliminated from the study because of unsatisfied DNA extraction.The total positive rate of HPV DNA detected by PCR in cervical adenocarcinoma and adenosquamous carcinoma was 91.8%(56/61).Using general primer GP5/GP6,the positive rate was 50.8%(31/61).Using SPF primers for the 30 negative cases,25 additional HPV positive cases were founded.All the positive samples had at least one of the high risk types.HPV16 was the most preferential type followed by HPV18,31,39 and 45.The infection of HPV 16 and 18 accounted for about half of HPV-positive adenocarcinoma.Multiple HPV infections were found in 21.4%(12/56)of the cases.Conclusion:The high risk type of HPV is associated with cervical adenocarcinoma.Single infection is more frequently presented than multiple infections,no single type of HPV plays a predominated role even HPV16 and 18 are the major types.