人类乳头瘤病毒HPV16,18型感染与胃癌发生的关系

目的探讨人类乳头瘤病毒ＨＰＶ１６,１８型感染与胃癌发生的关系．方法采用ＨＰＶ１６Ｅ６和ＨＰＶ１８Ｅ６／Ｅ７特异性引物,通过多聚酶链反应（ＰＣＲ）,检测３７例新鲜胃癌组织标本及２０例新鲜癌旁正常胃粘膜标本中的ＨＰＶＤＮＡ．结果新鲜胃癌组织３７例中ＨＰＶ１６,１８型ＤＮＡ检出率分别为２１６％和５４％．新鲜癌旁正常胃粘膜标本２０例中检出率为０．ＨＰＶ１６型ＤＮＡ在胃癌和癌旁正常胃粘膜中检出率存在显著性差异（Ｐ＜００５）,而ＨＰＶ１８则没有显著性差异（Ｐ＞００５）．新鲜胃癌组织３７例中ＨＰＶ１６,１８型ＤＮＡ总检出率为２７０％,２０例新鲜癌旁正常胃粘膜标本中总检出率为０,两者有显著性差异（Ｐ＜００５）．新鲜胃癌组织３７例中ＨＰＶ１６,１８型双重感染检出率为０．ＨＰＶ１６和（或）ＨＰＶ１８型ＤＮＡ的检出率在不同分化程度的胃癌中无显著性差异,但ＨＰＶ１６型ＤＮＡ在不同部位的胃癌中检出率却有显著性差异,以贲门部胃癌检出率最高．结论高危类ＨＰＶ１６型感染可能与胃癌发生有关,ＨＰＶ１６型病毒癌基因可能在胃癌形成过程中起一定作用     

胃幽门螺杆菌感染与胃癌发生关系的探讨

采用ＰＣＲ技术对５６例胃癌、６０例慢性胃炎及１２例正常人的胃粘膜组织检测了幽门螺杆菌（ＨＰ）。结果胃炎ＨＰ检出率９３．３３％，胃癌４１．０７％，正常人１６．６７％。提示胃炎的发生与ＨＰ感染明显相关，胃癌发生与ＨＰ感染有一定关系。贲门癌ＨＰ感染率（９．５２％）明显低于胃窦部及小弯区癌（６８％，３７．５％）。高分化型（腺样型）癌组织中ＨＰ感染率（５５．８８％）较低分化型（弥漫型１０％）高。     

结直肠癌组织中人乳头瘤病毒DNA的研究

目的研究１６、１８型人乳头瘤病毒（ＨＰＶ）是否与结直肠癌的发生有关．方法结直肠粘膜活检组织１２３例，其中结直肠癌３５例，乳头状腺瘤１７例，炎性息肉１１例，结肠炎３０例，以及正常结肠粘膜３０例，用对各型ＨＰＶ高度保守的通用引物和１６、１８型特异性引物作聚合酶链反应（ＰＣＲ）检测ＨＰＶＤＮＡ．结果ＨＰＶＤＮＡ总检出率１４６％，正常粘膜，结肠炎和炎性息肉组为３３％（２／７１），乳头状腺瘤为１７６％（３／１７），结直肠腺癌为３７１％（１３／３５）．在正常粘膜，结肠炎和炎性息肉组未发现１６、１８型ＨＰＶＤＮＡ，在乳头状腺瘤组有３例为１８型ＨＰＶＤＮＡ阳性，结直肠腺癌组１３例ＨＰＶ阳性病例中有３例为１６型，９例为１８型感染；在正常组织、癌旁组织和癌组织中ＨＰＶＤＮＡ检出率依次增高．ＨＰＶ在直肠、左半结肠，右半结肠感染率依次为２８６％，１４％，２６％．结论结肠癌的发生与ＨＰＶ１６、１８型有关，腺癌以１８型感染为主．ＨＰＶＤＮＡ检出率在右半结肠，左半结肠和直肠依次增高．     

胃癌组织P16，P53蛋白和增殖细胞核抗原的表达意义

目的探讨Ｐ１６,Ｐ５３蛋白和增殖细胞核抗原（ｐｒｏｌｉｆｅｒａｔｉｎｇｃｅｌｎｕｃｌｅａｒａｎｔｉｇｅｎ,ＰＣＮＡ）在胃癌的发生、发展中的作用及临床意义．方法应用免疫组织化学方法,对７７例胃癌和癌旁粘膜组织、２１例胃正常组织中Ｐ１６,Ｐ５３蛋白表达产物和ＰＣＮＡ进行检测,并结合临床资料进行分析．结果胃癌组织中Ｐ１６蛋白的阳性率为２０８％（１６／７７）,明显低于癌旁粘膜组织５９７％（４６／７７）和胃正常组织９０５％（１９／２１,Ｐ＜００５）;Ｐ５３蛋白与ＰＣＮＡ在胃癌组织中的阳性率为８０５％（６２／７７）和９２２％（７１／７７）,明显高于癌旁粘膜组织４１６％（３２／７７）,６４９％（５０／７７）和胃正常组织００％（０／２１,Ｐ＜００５）．Ｐ１６,Ｐ５３蛋白和ＰＣＮＡ阳性表达与胃癌组织学类型、浸润深度、分化程度及淋巴结转移有显著性差异（Ｐ＜００５）,与患者年龄、性别、肿瘤大小及部位无关（Ｐ＞００１）．结论Ｐ１６,Ｐ５３蛋白和ＰＣＮＡ的异常表达对胃癌的发生发展、恶性程度、淋巴结转移及预后有密切关系和重要临床意义．     

人乳头瘤病毒与大肠癌的发生及病理组织学相关性的研究

目的 了解人乳头瘤病毒（ＨＰＶ）与大肠癌的发生及病理组织学的关系。方法 用多重引物ＰＣＲ方法检测了４６例大肠癌标本和３６例对照组织的ＨＰＶ ＤＮＡ,并与病理组织９学结果进行比较。结果 大肠癌总ＨＰＶ阳性率为４１．３％,其中ＨＰＶ１６为２３．９％,与对照组比较,差异有显著性（Ｐ〈０．０５）。总ＨＰＶ和ＨＰＶ１６的阳性纺,高分化腺癌组为３８．９％和１６．７％,中分化组为３８．５％和２３．１％,低分化组     

不同病理类型胃息肉的细胞DNA定量研究

为探讨不同病理类型胃息肉的ＤＮＡ生物学行为及其与胃癌的关系，应用流式细胞术对５７例正常胃粘膜、胃息肉和胃癌组织作细胞ＤＮＡ定量分析。正常胃粘膜、炎性息肉、增生性息肉、腺瘤性息肉及胃癌中，ＤＮＡ非整倍体检出率分别为０％、７．７％、９．１％、３６．４％和５８．３％。各型胃息肉和胃癌细胞增殖指数（ＰＩ）均显著地高于正常胃粘膜（Ｐ＜０．０１）；增生性息肉组和炎症性息肉组ＰＩ低于胃癌组，差异有显著性（Ｐ＜０．０５）或非常显著性意义（Ｐ＜０．０１），而腺瘤性息肉组ＰＩ与胃癌组近似（Ｐ＞０．０５）。结果表明胃息肉是一种细胞增殖活跃性病变，其中腺瘤性息肉的ＤＮＡ生物学行为更接近于胃癌，可能容易癌变。     

以地高辛标记探针研究人16型乳头瘤病毒感染与良性前列腺增生症的相关性

目的用分子生物学方法制备地高辛标记１６型人乳头瘤病毒（ＨＰＶ１６）ＤＮＡ探针，对ＨＰＶ与良性前列腺增生症（ＢＰＨ）的相关性进行探讨。方法将质粒ｐＨＰＶ１６经ＢａｍＨＩ酶切，Ｖ－形槽电洗脱回收ＨＰＶ１６ＤＮＡ，随机引物法进行标记。用斑点杂交对探针的质量参数进行评估，并对２９例ＢＰＨ和７例正常前列腺组织进行了ＨＰＶ１６ＤＮＡ检测。结果表明本探针灵敏特异、经济实用。ＢＰＨ组和正常对照组的阳性率分别为６２．１％和１４．３％，两者差异有显著性（Ｐ＜０．０５）。结论ＢＰＨ可能是ＨＰＶ感染的又一男性宿主，ＨＰＶ１６感染可能在ＢＰＨ中起着一定的病因学作用。     

大肠肿瘤组织中人乳头状瘤病毒DNA的分子原位杂交研究

目的探讨ＨＰＶ１６和ＨＰＶ１８感染与大肠肿瘤发生的关系．方法采用地高辛标记的ＨＰＶ１６和ＨＰＶ１８ＤＮＡ探针分别在４０例大肠癌和３０例大肠腺瘤组织石蜡切片上进行原位杂交探测ＨＰＶＤＮＡ．结果受检大肠腺瘤组织ＨＰＶＤＮＡ阳性８例（２７％），其中ＨＰＶ１６ＤＮＡ５例，ＨＰＶ１８ＤＮＡ３例，主要见于管状绒毛状腺瘤和绒毛状腺瘤；大肠腺癌组织中ＨＰＶＤＮＡ阳性１９例（４８％），其中ＨＰＶ１６ＤＮＡ１４例，ＨＰＶ１８ＤＮＡ５例．ＨＰＶＤＮＡ主要见于肿瘤细胞核中，少部分见于胞质中．研究表明，大肠癌组织中ＨＰＶ１６，１８ＤＮＡ检出阳性率明显高于大肠腺瘤；且大肠绒毛状腺瘤中ＨＰＶＤＮＡ阳性率明显高于管状腺瘤，阳性率接近高分化腺癌．结论ＨＰＶ１６，１８型感染并整合至宿主细胞ＤＮＡ中与肠癌的发生有密切的关系     

肺癌患者人乳头瘤病毒16,18型,P53蛋白及端粒酶活 …

目的 探讨人乳头瘤病毒１６、１８（ＨＰＶ１６、１８）型感染,Ｐ５３蛋白表达,端粒酶活性在不同民族原发性肺癌组织中是否存在差别及其意义。方法 分别应用聚合酶链反应（ＰＣＲ）、免疫组化技术测定１１０例肺癌组织中ＨＰＶ１６、１８－ＤＮＡ、Ｐ５３蛋白表达,用以ＰＣＲ为基础的端粒重复扩增法（ＰＣＲ－ＴＲＡＰ）测定其中２２例肺癌新鲜组织内端粒酶活性。结果 肺癌组ＨＰＶ１６、１８总阳性率为４０．０％,与正常对照     

胃粘膜p53抑癌基因异常与癌变的研究

目的系统研究各种胃粘膜病变中ｐ５３抑癌基因的变异和意义．方法内镜活检组织１５４例，其中浅表性胃炎３０例，萎缩性胃炎３３例，萎缩性胃炎伴肠上皮化生３１例，萎缩性胃炎伴异型增生３０例，胃腺癌３０例．用免疫组化法检测Ｐ５３蛋白表达；用单链构象多态性分析及ＤＮＡ测序检测ｐ５３第５～８外显子点突变．结果胃良性病变粘膜未见Ｐ５３蛋白表达．胃癌中Ｐ５３蛋白阳性表达率为３３３％（１０／３０例）．单链构象多态性分析ｐ５３点突变的检出率，在胃癌为５４５％（６／１１例），异型增生为２０％（３／１５例），肠化生为６７％（１／１５例），测序证实１例胃癌在第５外显子存在点错义突变和碱基缺失，２例异型增生在第６外显子存在点错义突变．结论ｐ５３抑癌基因变异与胃粘膜癌变有关．     

Evidence of human papilloma virus infection and its epidemiology in esophageal squamous cell carcinoma

AIM: To look for the evidence of human papilloma virus (HPV) infection in esophageal squamous cell carcinomas (ESCC) and to investigate the potential role and epidemiology of HPV infection in the pathogenesis of esophageal carcinomas in Henan emigrants. METHODS: Papilloma virus (PV) and HPV were determined by UltrasensiveTM S-P immunohistochemistry (IHC) and in situ hybridization (ISH) in esophageal carcinoma tissues (82 cases) and the normal mucosa (40 cases). RESULTS: IHC revealed that the positive rate of PV was 75.0%, 68.18% and 72.5% respectively while the HPV (16/18-E6) positive rate was 45.0%, 36.36%, 37.5%, respectively in esophageal carcinoma tissue specimens from Henan emigrants,the local citizens and patients in Hubei Cancer Hospital. The PV and HPV (16/18-E6) were negative in all normal esophageal mucosa specimens. No correlation was found between HPV in esophageal squamous cell carcinoma tissues and in grade 1-3 esophageal squamous cell carcinoma cells. In situ hybridization showed that the HPV (16/18) DNA positive rate was 30.0%, 31.8%, 25.0%, respectively in the 3 groups of samples. No positive hybridization signal was found in 40 normal esophageal mucosa specimens. The positive rate of HPV (16/18) DNA in the esophageal carcinoma specimens was significantly higher than that in normal mucosa specimens (P<0.05). The positive rate was not different among the 3 groups of esophageal carcinoma tissue specimens(P>0.05). CONCLUSION: HPV infection is high in esophageal carcinoma of Henan emigrants, local residents and patients in Hubei Cancer Hospital. HPV is closely related with esophageal squamous cell carcinoma. HPV infection may play an important role in esophageal squamous cell carcinoma.     

Detection of human papillomavirus type 16 DNA in formalin-fixed, paraffin-embedded tissue specimens of gastric carcinoma

OBJECTIVES: Human papillomavirus type 16 (HPV16) is regarded as one of the important tumor-related viruses, which are known to have a role in cervical carcinoma; however, there are few reports on HPV16 in gastric carcinoma (GC). Our study aimed to investigate the relationship between HPV16 and the occurrence of GC. METHODS: Liquid PCR (LPCR) and in-situ PCR (ISPCR) methods were carried out to detect the HPV16 oncogene E6 cell-type-specific enhancer in the long control region of HPV16 in 40 GCs and corresponding gastric adjacent normal mucosa (GANM). The patients were from Shaanxi Province in China; Helicobacter pylori (Hp) was detected by immunohistochemistry and by hematoxylin and eosin staining in their gastric tissues. RESULTS: The HPV16 E6 gene was detected in 37.5% (15/40) of the GCs and 5% (2/40) of the GANMs with LPCR, as was the cell-type-specific enhancer; however, the positive rate of E6 was 27.5% (11/40) in GCs and 0% (0/40) in GANMs, respectively, with ISPCR. HPV16 DNA was mainly located in the nucleus of gastric glandular epithelium cells. The infection rate of HPV16 DNA in GCs was higher than that in GANMs (P=0.0004), and the HPV16 had no statistical correlations with sex, age, invasion, grading or lymph node metastasis (P>0.05). The infection rate of HPV16 in cardiac GCs was significantly higher than that in noncardiac ones (P=0.0136), and HPV16 had no correlation with Hp in GCs (P=0.0829). Receiver operator characteristic curve analysis indicated that there was no statistical difference between the LPCR and ISPCR methods in our study through optimizing parameters in ISPCR procedures (P=0.768). CONCLUSIONS: These findings suggested that HPV16 can infect gastric glandular epithelium cells and that viral infection might play a role in the occurrence of GCs independent of or without the cooperation of an Hp infection.     

食管癌组织中人乳头状瘤病毒感染状况分析

为探讨人乳头状瘤病毒（HPV）感染在食管癌发病中的作用，采用荧光定量聚合酶链反应对河北省高发区食管癌、癌前病变、正常食管组织和低发区食管癌组织中人乳头状瘤病毒（HPV16、18）进行检测。结果显示，高发区食管癌组织HPV16、18DNA阳性率为38.7%，癌前病变为25％，正常食管组织为2.5%，低发区食管癌组织为20％。高发区食管癌和癌前病变与正常食管组织HPV16、18感染率相比差异具有显著性（P＜0.01）；低发区食管癌组织HPV16、18感染率与高发区正常食管组织相比差异具有显著性（P＜0.01）；低发区食管癌组织HPV16、18感染率低于高发区食管癌组织，但差异无显著性（P＞0.05）。提示河北省食管癌高低发地区食管癌组织、高发区食管癌前病变组织中均存在较高的人乳头状瘤病毒感染率，为食管癌发病的重要因素之一。     

胃黏膜癌变过程中端粒长度变化及其与细胞内DNA含量的关系

目的　分析不同病变胃黏膜细胞内端粒长度的差异,以及细胞内DNA含量,探讨端粒行为异常、细胞内DNA含量与胃黏膜癌变的关系。方法　应用Southern杂交分析细胞内端粒长度,应用流式细胞术测定细胞内DNA含量。结果　在172例胃镜活检标本中,正常胃黏膜,慢性浅表性胃炎(CSG),伴0、1、2度肠化的慢性萎缩性胃炎(CAG)和胃癌组织的端粒长度,分别是(10.42±0.2)kb,(9.86±0.4)kb,(9.78±1.2)kb、(8.64±1.0)kb、(6.22±1.2)kb和(5.86±2.6)kb。45例胃癌手术标本结果　相似。流式细胞术分析细胞内DNA含量,在胃镜活检标本中,正常胃黏膜,CSG,伴0、1、2度肠化的CAG和胃癌组织的异倍体DNA检出率分别为0、0、0、10.00％、12.50％6和33.33％6。45例胃癌手术切除标本结果　相似。异倍体细胞内端粒长度明显短于二倍体细胞,异倍体细胞中端粒长度与DNA指数呈负相关(r=-0.91,P＜0.01),即端粒越短,DNA指数越高。结论　端粒长度从正常胃黏膜、不同程度肠化胃黏膜到癌变胃黏膜逐渐缩短。在正常胃黏膜和CSG中未检出异倍体DNA,从1度、2度肠化到癌变胃黏膜异倍体DNA检出率逐渐增高,而在异倍体细胞中端粒长度和DNA指数呈负相关。推测,可能存在端粒愈短,DNA扩增愈活跃。端粒缩短伴DNA指数增加可能是胃癌发生的先兆。     

Detection of human papillomavirus DNA in squamous cell carcinoma of the esophagus by auto-nested PCR

The aim of the present study was to investigate the presence of human papillomavirus (HPV) in surgical specimens of esophageal squamous cell carcinoma. One hundred and sixty-five paraffin-embedded specimens of esophageal carcinoma were analyzed through high-sensitivity auto-nested polymerase chain reaction (PCR) using the consensus GP5+/GP6+ primer. Twenty-six specimens of esophageal mucosa without malignant disease were also studied as a control group. Two different specific primer sets targeting the E6 region of the HPVs 16 and 18 were used for typing. Direct DNA sequence analysis was conducted to confirm positive PCR results. HPV DNA was detected in 26 esophageal carcinomas (15.75%), but in none of the benign esophageal specimens (P < 0.05). Out of the 26 positive cases, 24 were HPV-16 and one was HPV-18. One tumor contained both HPV-16 and -18 DNA. Positive PCR results were confirmed by the amplified viral sequences. Our findings suggest that the presence of either HPV-16 or -18 might be related to development of the malignant phenotype in the esophagus.     

Prevalence of human papillomavirus in esophageal carcinoma in Tangshan,China

AIM:to study the prevalence of human papillomavirus(HPV) in esophageal carcinoma in tangshan,China,a high-incidence area.METHODS:Formalin-fixed,paraffin-embedded tissue specimens from 198 patients who were pathologically diagnosed with esophageal squamous cell carcinoma from 2011 to 2013 were obtained from a pathology department in Tangshan.DNA was extracted from all198 specimens to detect HPV by polymerase chain reaction(PCR).β-globin PCR was performed to check the quality of the DNA extraction procedure.PCR was performed to detect a wide range of HPV types,and type-specific PCR was performed to detect HPV types16 and 18.Negative and positive controls were used for HPV 16 and 18 detection.RESULTS:the DNA extraction method in this study appeared to be more effective than other previously reported methods.After DNA extraction,more than98%of the tissue specimens had an acceptable result in the DNA qualification test(β-globin PCR).the overall prevalence of HPV in tumor tissues by GP6+/GP5+PCR was 79.79%,and the prevalence of HPV types16 and 18 was 40.40%and 47.47%,respectively.PCR demonstrated the presence of HPV,and direct sequencing confirmed the HPV genotypes.All HPVpositive PCR products were checked by DNA sequence analysis using DNAman and compared with the known HPV sequences listed in the basic Local Alignment Search tool database to evaluate the HPV types.this analysis confirmed the presence of HPV types 16 and18.CONCLUSION:DNA of high-risk HPV types 16 and 18is present in esophageal tumors,implicating HPV as a possible etiologic factor for esophageal squamous cell carcinoma.     

人乳头病毒16、18型与人胃腺癌关系的探讨

目的：采用ＰＣＲ法检测胃癌患者人乳头病毒（ＨＰＶ）１６，１８型ＤＮＡ序列，探讨ＨＰＶ类型与人胃腺癌的关系，为今后进行胃癌基因治疗及预防提供理论及实验依据。     

Human papillomavirus involvement in esophageal precancerous lesions and squamous cell carcinomas as evidenced by microscopy and different DNA techniques.

A series of 71 surgically resected esophageal squamous cell carcinomas, including 51 cases of formalin-fixed samples and 20 cases of fresh biopsy specimens derived from the high-incidence area of esophageal cancer in China, were systematically analyzed for the presence of human papillomavirus (HPV) infections by light microscopy, electron microscopy (TEM), in situ DNA hybridization, Southern blot hybridization, and polymerase chain reaction (PCR) techniques. On light microscopy, HPV-suggestive lesions were found in a total of 49.0% (25 of 51) of the specimens, including the flat type (22 of 51) and, less frequently, an inverted one (2 of 51). Of the 51 formalin-fixed, paraffin-embedded specimens, 43.1% (22 of 51) contained HPV DNA sequences by in situ hybridization. Of the positive cases, HPV 6 was present in three (5.9%), HPV 11 in three (5.9%), HPV 16 in eight (15.7%), HPV 18 in six (11.8%), double infections with HPV 11/18 in one (2.0%), and HPV 16/18 in one. In most cases the HPV-positive signals were localized in the hyperplastic and/or dysplastic epithelium adjacent to invasive carcinomas. In two specimens, however, HPV DNA sequences were found in the frankly invasive lesions, one being HPV 6 and the other HPV 18. On TEM, HPV-like particles located in the nuclei of koilocytotic cells were demonstrated in two of the five specimens previously shown to be HPV-positive by in situ hybridization. By means of the PCR technique, all specimens positive for HPV by in situ hybridization also contained amplified HPV sequences. Moreover, three additional samples negative by in situ hybridization were found to contain HPV 11 DNA sequences. Of the 20 DNA samples extracted from the fresh carcinoma samples (containing some surrounding tissues as well) 9 were shown to contain HPV DNA sequences by Southern blot hybridization under low-stringency conditions. Of these, eight samples remained positive when hybridized with the probe cocktail of HPV 11, 16, 18, and 30 DNA under high-stringency conditions. HPV DNA sequences in these carcinoma specimens appeared to be present mainly in an integrated form. The present results confirm the HPV involvement in esophageal squamous cell lesions and suggest that HPV infection might be an important etiologic factor in the pathogenesis of esophageal cancer, most probably acting synergistically with other carcinogenic factors.     

Human papillomavirus DNA and P16(INK4A) expression in concurrent esophageal and gastric cardia cancers

AIM:To investigate the relationship between human papillomavirus (HPV) infection and concurrent esophagus and gastric cardia cancer from the same patient (CC) and examine the significance of P16 INK4A protein expression.METHODS:Polymerase chain reaction was used to detect the presence of HPV type16 (HPV16).The expression of P16 INK4A protein was detected using immunohistochemistry.RESULTS:Among the CC specimens,HPV16-DNA was found in eight cases of esophageal squamous cell carcinoma (ESCC) and five cases of gastric cardia adenocarcinoma (GCA),respectively (47% vs 29%),and two of both ESCC and GCA.P16 INK4A was highly expressed in both ESCC and GCA.In the HPV-associated positive CC,higher P16 INK4A expression was observed in the GCA than in the ESCC (75% vs 25%,P < 0.05).CONCLUSION:HPV16 as a correlated risk factor may play an important role in the development of ESCC and GCA.P16 INK4A may be a screening index in the HPVassociated carcinoma of gastric cardia.