FHIT基因在肺癌中的缺失与HPV感染的研究

目的　探讨肺癌发生的分子生物学机制。方法　采用逆转录 -巢式聚合酶链反应 (reverse tapepolymerasechainreaction)的方法对 42例肺癌及 10例正常肺组织中的FHIT(Fragilehistidinetriad)基因的缺失情况进行检测 ,并用PCR技术检测了肺癌组织中人乳头状瘤病毒 (humanpapillomavirus ,HPV)的DNA片段。 结果　 66.7% (2 8/4 2 )肺癌组织中检测到FHIT基因的缺失 ,而正常组织中未检测到FHIT基因的缺失 ,二者差别有显著意义 (P <0 .0 1)。 42例肺癌组织中有 8例检测到HPV的片段 ,阳性率为 19% (8/4 2 ) ,正常组织中未检测到HPV的片段 ,且在 8例HPV阳性的标本中均有FHIT基因缺失。结论　 (1)FHIT基因的缺失在肺癌的发生中起一定的作用。 (2 )FHIT基因的缺失与肺癌的不同病理分型、分化程度、临床分期无关。与吸烟有一定的相关性。 (3 )HPV的感染与肺癌的发生有一定关系 ,且与FHIT基因缺失有正相关关系。     

PCR-LIS-SSCP检测大肠癌PTEN基因的点突变

目的　探讨PTEN基因突变在大肠癌发病机理中的作用。方法　应用聚合酶链反应 -低离子强度 -单链构象多态性技术 ,对大肠癌及正常大肠石蜡包埋组织中PTEN的第 7、第 8个外显子进行点突变检测。结果　在 60例大肠癌组织中 ,第 7个外显子SSCP带型异常的有 2 8例 ,阳性率为 46.7% (χ2 =5 .0 81,P <0 .0 5 ) ,第 8个外显子未见阳性带。在 10例正常大肠组织中 ,2个外显子均未见异常SSCP带型。结论　大肠癌组织中PTEN的第 7个外显子点突变的发生率高 ,PTEN基因突变可能在大肠癌发病机理中有重要作用     

大肠癌患者粪便基因检测的研究进展

人类结直肠粘膜上皮更新速度快,脱落的上皮细胞随粪便排出体外,检测脱落细胞的癌基因及其表达产物,将有助于大肠癌的早期诊断,且检测方法无创,依从性好,有广阔的应用前景。现对粪便中结直肠瘤相关突变基因检测的研究进展作文献回顾。     

应用 PCR 方法检测大肠癌组织中的 HPV DNA

本文采用聚合酶链式反应（ＰＣＲ）的方法，对２０例大肠癌患者的手术切除标本进行了人体乳头瘤病毒（ＨＰＶ）ＤＮＡ的检测。结果发现４例（２０％）ＨＰＶ１６型扩增阳性，其中１例显示ＨＰＶ１６．１８型双重阳性。初步表明大肠癌组织中存在ＨＰＶ的感染。     

新疆维吾尔族妇女宫颈癌组织中HPV16型E6基因突变分析

背景与目的：高危型人乳头状瘤病毒16和18(human papillomavirus type16 and 18,HPV16,HPV18)是宫颈癌主要病因之一,尤其以HPV16最为常见,其中HPV16E6是主要癌基因之一。在一些地区,特定的E6基因突变株是宫颈癌发生的危险因素。新疆南部维吾尔族聚居区足宫颈癌高发区,我们已在前期的研究中发现该地区HPV16E6基因发生突变。本研究旨在检测该突变在新疆南部维吾尔族妇女宫颈癌组织中的分布规律,并探讨其与该地区宫颈癌高发的关系。方法：从35例中国新疆南部维吾尔族妇女宫颈癌活检标本中提取组织DNA作为模板,PCR扩增HPV16E6全长基因,PCR产物直接测序或克隆后测序,分析新疆维吾尔族妇女宫颈癌组织中HPV16E6基因的突变。结果：PCR检测结果表明宫颈癌组织中HPV16E6阳性率为82．86％(29／35);26例中E6分离片段的测序和序列分析表明,15例(57．69％)分离株E6基因与原型相同,另有11例(42．31％)E6基因突变,其中9例(34．62％)分离株发生了L83V突变,2例(7．69％)分离株发生rL83V／D63E突变。结论：中国新疆南部地区HPV16E6基囚发生变异,其原型和变异型在该地区维吾尔族宫颈癌患者巾的分布规律可能与该地区宫颈癌高发存在一定关系。     

HPV16E7 siRNA表达载体抑制宫颈癌SiHa细胞E7基因的研究

目的：研究HPV16E7siRNA表达载体对宫颈癌SiHa细胞E7基因的抑制作用。方法：利用脂质体将HPV16E7 siRNA表达载体psiRNA-1、psiRNA-2、psiRNA-3及空载体psiRNA转染SiHa细胞,以荧光定量RT-PCR和流式细胞仪检测不同时间点E7 mRNA和蛋白的变化。结果：载体psiRNA-1、psiRNA-2和psiRNA-3均能抑制SiHa细胞E7基因 mRNA和蛋白的表达,其中载体psiRNA-1的抑制作用最强。在抗性克隆形成后1周和4周,对E7mR-NA和蛋白抑制率分别为92.15%、84.30%和65.69%、59.11%。而空载体对E7mR-NA和蛋白的表达均无明显抑制作用。结论：HPV16E7 siRNA表达载体能较长期地抑制宫颈癌SiHa细胞E7基因的表达。     

子宫颈癌及癌前病变组织中p53基因产物表达与HPV感染的关系

目的探讨HPV感染和宫颈癌及癌前病变组织中p53基因产物的表达及临床意义。方法应用原位标记(PRINS)技术和S-P法对61例宫颈癌,42例CIN,15例正常宫颈组织进行HPV16检测和p53基因产物的表达。结果宫颈癌组和CIN组的HPV16感染率明显高于正常宫颈组(P<0.01);p53基因产物阳性表达率宫颈癌组为47.5%,高于正常宫颈组(P<0.05);p53和HPV16有相关性。结论HPV感染和p53基因产物的表达与宫颈癌的发生、发展及转移密切相关,HPV可能通过造成p53基因突变失活,导致宫颈癌的发生。     

宫颈癌患者外周血中HPV6.11、HPV16.18、HSVⅡ及HCMV DNA的检测

目的：测定宫颈癌患者外周血中人乳头状瘤病毒（HPV）、单纯疱疹病毒（HSVⅡ）和人巨细胞病毒（HCMV）DNA，探讨其对HPV感染的高危妇女发生宫颈癌的追踪及监测作用。方法：采用HPV6．11、HPV16．18、HSVⅡ及HCMV PCR试剂盒，用PE Gene Amp PCR System 2400扩增，测定104例宫颈癌患者外周血中的HPV6．11、HPV16．18和80例HSVⅡ及HCMV的DNA，并以32例子宫肌瘤患者为对照组。用χ^2行差异性检测。结果：1）宫颈癌患者外周血中HPV6．11、HPV16．18、HSVⅡ和HCMV病毒DNA测定的阳性率分别为35．58％、20．19％、10．00％和3．75％。对照组分别为12．50％、9．37％、3．13％和0％。2）宫颈癌患者外周血中HPV6．11混合HSVⅡ或混合HCMV DNA阳性率分别为2．04％和1．92％；HPV16．18混合HSVⅡ或混合HCMV DNA阳性率分别为5．36％和3．30％。对照组无1例出现病毒DNA的混合阳性。结论：1）宫颈癌患者外周血中HPV、HSVⅡ和HCMV等病毒DNA阳性率高于对照组。2）宫颈癌患者外周血中的病毒DNA阳性率以HPV为主（P＜0．05）。3）宫颈癌患者外周血中各种病毒DNA混合阳性率较低，其协同的致癌作用有待进一步研究。4）在HPV感染的高危人群中定期进行宫颈涂片和检测外周血中HPV DNA，可起到对病情的追踪及监测作用。     

hWAPL基因在宫颈癌中作用的研究进展

hWAPL（human wing—apart like）基因是果蝇体内WAPL基因的同源基因。hWAPL在宫颈癌组织中显著表达，且与癌变的严重程度显著相关。人乳头状瘤病毒（HPV）癌蛋白E6和E7可诱导hWAPL的表达。文章主要综述hWAPL基因在宫颈癌的发生发展中的作用。     

Presence of Human Papillomavirus DNA in Colorectal Cancer Tissues in Shiraz,Southwest Iran

Background: Colorectal cancer is one of the most common cancers worldwide. Viruses including human papillomavirus (HPV) have been reported to be associated with different cancers but any association with colorectal cancers remains controversial. Aim: To evaluate any association between HPV infection and adenocarcinoma of the colon and adenomatous polyps. Materials and Methods: Paraffin-embedded tissue specimens of 70 colorectal adenocarcinomas, 70 colorectal adenomatous polyps, and 70 colorectal normal tissues were subjected to DNA extraction. The quality of the extracted DNA was confirmed by amplification of a β-globin fragment using polymerase chain reaction (PCR). PCR using specific primers were performed to detect HPV DNA. Specific primers targeting the E6 region of the HPVs 16 and 18 were used for genotyping. Results: HPV DNA was detected in 2 (2.85%) out of 70 adenocarcinoma colorectal tissues and 4 (5.71 %) out of 70 adenomatous colorectal tissues. All normal colorectal tissues were negative for HPV DNA. HPV-16 was the most predominant genotype (5 sample) followed by HPV-18 (4 sample). Despite the above observations, statistical analyses indicated no significant differences in the frequencies of HPV positive subjects between the cancerous and normal samples. Conclusions: Although the differences observed in the frequencies of HPV positive cases in our study was not significant relative to those of control subjects, the fact of 6 positive samples among cancerous tissues, may still suggest a role of HPV in colorectal carcinogenesis. The study collectively indicated that some colorectal cancerous tissues are infected with high risk HPV genotype. The findings merit more investigation.     

人类乳头状瘤病毒与宫颈糜烂癌变的相关性研究

目的：探讨人类乳头状瘤病毒与宫颈糜烂和癌变的相互关系。方法：阴道镜下对116例宫颈糜烂患者、12例宫颈癌患者及36例正常妇女宫颈，分别钳取活组织；用聚合酶链反应，对活组织进行HPV公共引物及HPV16、HPV18型特异引物检测。结果：宫颈糜烂患者HPV阳性率为29．31％，正常妇女宫颈HPV阳性率仅11．11％（P＜0．05），宫颈颗粒型或乳突型糜烂与宫颈癌强相关（OR＝5．56％CI＝1．21－25．459）。“高危”HPV－DNA检出率随宫颈鳞柱上皮异位程度增加而升高。结论：宫颈糜烂癌变的病因可能与持续感染“高危”HPVs有关。     

The Prevalence of Human Papillomavirus in Cervical Cancer in Iran

Background: The human papiloma virus (HPV), which is sexually transmitted, and most commonly causesgenital warts, has been linked to cervical intraepithelial neoplasia and invasive carcinoma. Of ninety plus types ofHPV, HPV-16 is the most prevalent in cervical cancer, followed by HPV-18, and HPV-33. As HPV’s implication hasnot been assessed in the Middle East the main focus of this retrospective study was to determine the prevalence ofHPV –16,18, and 33 in cases of cervical cancer from Iran.Material and Methods: This retrospective study covered 100 patients with uterine cervical carcinomas who werereferred to two referral centers for cancer in Tehran-Iran. Pathological blocks were collected for these cases andinitial review of the blocks showed poor specimens in 18 cases, which left 82 cases for the study. These samples werehistologically examined to verify the presence and the type of carcinoma. The next step was in situ hybridzation forthe detection of HPV common DNA. In Situ hybridization was preformed on all samples. Finally, Polymerase ChainReaction (PCR) was preformed for the HPV types 16, 18, and 33. PCR amplification of exon 5 of the p53 gene wasused as an internal control for the integrity of DNA. Takara PCR Human papilloma Detection method was usedwhich includes primer for HPV 16, 18, and 33. Three primers were used alone, or in combination, in order toincrease the sensitivity of the detection.Results: The majority of tumors were squamous cell carcinomas (87%). The rest were adenosquamous carcinomaand adenocarcinomas. None of the 82 different cervical carcinoma tissue samples were found to be positive by in situhybridization. In the PCR samples, amplification of DNA was observed for 69 tumor specimens. In the remainning13cases, the DNA in fixed tissue was degraded, as verified by the absence of an internal control band (p53). Out of thetotal 69 tumors (85.5%) with adequate DNA contained HPV band on PCR. The majority (73.9%) of HPV positivetumors contained HPV-16; the rest (11.6%) demonstrated type 18 and 33. There was no correlation between thehistology of carcinoma and presence of types of HPV.Conclusion: The prevalence of HPV in carcinomas of uterine cervix in Iran is similar to those reported in otherregions of the world. Similarly, it appears that HPV-16 is the most common type associated with cervical cancer inIran. Further studies on larger samples of patients, particularly in those with pre-invasive forms of the disease, areneeded to elucidate the carcinogenic role of HPV types in cervical cancer in Iranian women.     

Human Papillomavirus Genotypes and Cervical Cancer in Northeast Thailand

Human papillomavirus (HPV) is a major cause of cervical cancer. More than 100 HPV genotypes have beenidentified; however the distribution varies geographically and according to ethnicity. The purpose of this studywas to investigate the prevalence and distribution of HPV subtypes among Northeast Thai women. Subjectsincluded 198 cases of SCCA and 198 age-matched, healthy controls. HPV-DNA was amplified by PCR using theconsensus primers GP5+/6+ system followed by reverse line blot hybridization genotyping. The prevalence ofhigh-risk HPV infection was 21 (10.1%) and 152 (76.8%) in the controls and in the cases, respectively. High-riskHPV significantly increased the risk for cervical cancer with an OR of 42.4 (95%CI: 22.4-81.4, p<0.001) and anadjusted OR of 40.7-fold (95%CI: 21.5-76.8, p <0.001). HPV-16 was the most prevalent HPV type in the SCCA(56.2%) followed by HPV-58 (17.8%) and HPV-18 (13.6%); whereas HPV-58 (46.4%) was a prominent genotypein the controls followed by HPV-16 (39.3%) and unidentified HPV types (25.0%). These findings indicate thatHPV infection remains a critical risk factor for SCCA; particularly, HPV-16, HPV-58 and HPV-18. In orderto eradicate cervical cancer, sustained health education, promoted use of prophylactics and a HPV-58 vaccineshould be introduced in this region.     

Human Papillomavirus Infection and Risk Determinants for Squamous Intraepithelial Lesion and Cervical Cancer in Japan

A case control design was used to investigate human papillomavirus (HPV) prevalence and risk factors associated with development of cervical squamous intraepithelial lesion (SIL) and cervical cancer (CC) in Japan, One hundred and twenty-three women with histologically confirmed SIL or CC were compared to a control group of 778 cytologically normal women. With the use of a polymerase chain reaction (FCR)-hased method for detection of low-risk (types 6 and 11) and high-risk (types 16, 18, 31, 33, 35, 52 and 58) HPVs, a high prevalence of HPV infection was observed in smokers among the controls. Logistic regression analysis demonstrated that high-risk HPV infection was the most significant risk determinant for LSIL (OR=9.4, 95% CI=4.5–19), HSIL (OR=77, 95% CI=28–217) and CC (OR=97, 95% CI=35–269). It also showed that unmarried women, women married for 5 to 19 years and smokers represented high risk groups for SIL, while smokers and women with a history of many pregnancies/parities had increased risk for CC. Smoking was the only HPV infection-independent factor for CC, suggesting that smoking may have a carcinogenic effect on the cervix. Since neither history of other cancer nor family cancer history was associated with SIL or CC, genetic factors appear to play little role in cervical carcinogenesis. The risk for cervical neoplasia due to HPV infection increased after marriage in Japan, suggesting a role for husbands as carriers of HPV transmission. Protection from high-risk HPV infection may be of greatest importance for prevention of cervical cancer.     

江西地区妇女宫颈癌患者人乳头瘤病毒(HPV)感染各亚型分布特点

目的分析江西地区妇女宫颈癌患者人乳头瘤状病毒(human papillomavirus,HPV)感染分布情况及主要型别,为江西地区临床防治宫颈癌、研制适合该地区的预防性HPV疫苗提供理论依据。方法收集经病理确诊的941例宫颈癌患者治疗前宫颈脱落细胞,采用人乳头瘤病毒核酸扩增分型检测试剂盒方法,进行HPV分型检测。结果 941例宫颈癌患者中有780例HPV阳性,阳性率为82.9%,其中,单纯感染率为67.5%,混合感染率为15.4%,单纯高危感染率为66.6%。HPV16感染率最高为54.8%,宫颈癌中其他高危型的感染率从高到低依次为HPV58、18、52、31、33、39、53、45、51、68、59、66、56、35,未发现单纯HPV56感染。低危HPV型中HPV81型感染率最高为1.7%,其他低危型感染率从高到低依次为HPV44、6、42、11,未检测出低危型HPV43,且低危型HPV44仅存在于混合感染中。随着年龄增加,高危HPV16感染率呈逐渐下降趋势,差异有统计学意义(P=0.001,<0.05)。年龄≤50岁,HPV16感染率为59.5%,高于年龄>50岁HPV16感染率(50.7%),差异有统计学意义(P=0.007,<0.05)。年龄≤50岁HPV58感染率为6.7%,低于年龄>50岁HPV58感染率(11.7%),差异有统计学意义(P=0.019,<0.05)。而HPV亚型18、52、31分布均无差异。结论江西地区妇女宫颈癌患者中以HPV16、58、18、52感染为主要型别,HPV16感染是年轻宫颈癌的主要致病因素。混合感染并不增加宫颈癌的发生率。     

Evidence for an Association of Human Papillomavirus and Breast Carcinomas

Human papillomavirus (HPV) DNA has been detected in breast carcinoma by different laboratorial techniques, suggesting the virus could play a role in the pathogenesis of this tumor. The aim of the present study is to investigate the presence of HPV in patients with breast carcinoma and the correlation of the viral infection with prognostic factors for the disease outcome. Between June 2001 and July 2002, 101 paraffin embedded breast carcinoma specimens were analyzed through polymerase chain reaction (PCR) and sequencing of HPV-E6 gene. Twenty specimens of reduction mammoplasty and 21 specimens of fibroadenomas were also studied as a non-malignant control group. Two different specific primer sets targeting E6 region of the HPVs 16 and 18 were used for the analysis. The HPV DNA was detected in 25 breast carcinomas (24.75%), but in none of the benign breast specimens ( p < 0.001). Out of the 25 positive cases, 14 were HPV-16 positive (56%) and 10 were HPV-18 positive (40%). An original finding was the detection of both HPV-16 and -18 in a single tumor (4%). The amplified viral sequences confirmed the presence of HPV-16 and -18. No correlation between the presence of HPV DNA and specific prognostic predictors for the disease outcome was observed. Our results suggest that the presence in the breast of either HPV-16 or -18 might be related to development of the malignant phenotype. Further studies are warranted.     

遗传性非息肉病性大肠癌和普通遗传性大肠癌临床表型分析

目的:探讨遗传性非息肉病性大肠癌(HNPCC)与普通遗传性大肠癌临床表型的异同,为临床辨认HNPCC家系提供依据。方法:选择符合阿姆斯特丹标准Ⅱ或日本标准的22个HNPCC家系(A组)和普通遗传性大肠癌20个家系(B组)为研究对象进行随访分析。结果:1)A、B两组男女发病的比例分别为1.4:1(41/30)和1.5:1(38/26),无显著性差异(P>0.05)。2)A、B两组确诊的中位年龄分别为48岁(32～70岁)和61岁(30～83岁),50岁以前发病比例分别为59.2%和26.6%,A组较B组发病年龄明显提前,差异显著(P<0.01)。3)A、B两组右半结肠癌发病比例分别为56.9%(29/51)和29.2%(7/24),差异显著(P<0.05)。4)A组多发癌7例,B组未见多发癌。5)A组第一、二、三代的平均发病年龄分别为64岁、56岁、48岁,逐渐年轻化,B组无此现象。结论:HNPCC与普通遗传性大肠癌临床表型不尽相同,提示两者各自有其独特的遗传学特征。     

HPV16型感染与结直肠癌临床病理特征和预后关系的研究

 目的　探讨HPV16型DNA与结直肠癌临床病理和预后的关系。方法　结直肠癌50例、腺瘤38例和正常结直肠粘膜组织20例,应用多聚酶链反应(PCR)检测HPV16型DNA序列,并对其扩增产物进行电泳分析和southern blot分析。追踪观察患者的预后。结果　三组患者HPV16型DNA阳性率分别为42%、31.6%和0。结直肠癌和腺瘤分别与正常大肠粘膜组织比较,HPV16型DNA阳性率有显著性差异(P<0.05);结直肠癌和腺瘤之间比较,HPV16型DNA阳性率无显著性差异(P>0.05)。HPV16型DNA阳性与结直肠癌患者的年龄、性别,以及肿瘤的部位、大小、组织类型、临床分期均无关,但与其预后有关。结论　HPV16型感染是影响结直肠癌预后的因素之一。