Frequency of human papillomavirus infection in oesophageal squamous cell carcinoma in Iranian patients

Human papillomavirus (HPV) is 1 of the possible aetiological factors in the development of oesophageal squamous cell carcinoma (ESCC). We aimed to study the role of HPV in ESCC. 140 cases of ESCC were analysed for the HPV DNA by polymerase chain reaction (PCR) using GP5+/GP6+ primers for L1 open reading frame (ORF) to amplify a 150 bp segment of HPV L1 ORF. This region was subsequently sequenced to identify the type of HPV. Of the 140 patients enrolled to our study 50.7% were female and 49.3% were male, aged between 20 and 81 y. 33 tumour specimens (23.6%) and 12 (8.6%) non-involved tumour margins were HPV positive. From HPV positive tumour cases 36% were positive in tumour margins. The HPV positive cases were 21.7% male and 25.3% female. There is no correlation between presence and types of HPV with patients' gender and age. The frequency of HPV subtypes in tumoural regions was as follows: HPV-16, 60.6%; HPV-18, 30.3%; HPV-33, 6.1%; and HPV-31, 3%. We found only HPV-16 in tumour margins. Our results are consistent with HPV studies conducted in other high-risk areas for ESCC and provided further evidence to support a causal association of HPV infection with ESCC.     

人食管鳞状细胞癌标本中乳头状瘤病毒DNA的检测

目的食管鳞状细胞癌有明显的地域性差异,它的发生与遗传、环境、饮食和某些微生物的感染有密切的关系,本研究通过检测食管鳞状细胞癌和相应正常食管组织中HPV-11型和HPV-16型的DNA,阐明HPV与食管鳞状细胞癌的关系.方法用PCR的方法检测22位食管鳞状细胞癌患者手术切除的癌组织和相应癌旁正常食管粘膜(共44例标本)中的HPV-11型和HPV-16型DNA的存在情况,并对其中一例HPV阳性肿瘤标本的HPV序列进行测定. 结果在本组所涉及的22位患者共计44例标本中,对于HPV-11型,12例癌组织为阳性;有7例癌旁组织为阳性;癌组织和癌旁组织均为阳性的有6例.对于HPV-16型,6例癌组织为阳性;6例癌旁组织为阳性;癌组织和癌旁组织均为阳性的有3例.序列分析结果表明所测定的序列与GeneBank登录的NC001525.1(HPV-11)、M14119.1(HPV-11)和AF217526.1(HPV-11)的同源性极高,均为99%.结论通过实验结果分析,我们认为食管鳞状细胞癌与HPV-11型和HPV-16型主要衣壳蛋白L1基因密切相关,HPV可能在一定的地域环境中与食管鳞状细胞癌的发生具有相关性.马群风,男,1961-01-04生,贵州省贵阳市人,汉族.1983年遵义医学院医疗系本科毕业,1993年第四军医大学普胸外科专业硕士毕业.主要从事普胸外科疾病的临床和研究工作.发表论文4篇项目负责人马群风,710038,陕西省西安市,中国人民解放军第四军医大学唐都医院胸外科.     

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.     

Human papillomavirus detected in esophageal squamous cell carcinoma in Iran

IntroductionConsidering the different results obtained regarding the association of HPV with the development of esophageal squamous cell carcinoma (ESCC) in different populations, we aimed to determine the frequency of HPV infection and its subtypes in ESCC in Iranian patients.MethodA total of 100 paraffin-embedded tissue samples of ESCC diagnosed during 1991 and 2005 in the Institute of Cancer affiliated to Tehran University of Medical Sciences were selected. Seven out of 100 samples were excluded due to low quality of extracted DNA from paraffin-embedded specimens. Thus, 93 samples were included for HPV genotyping.ResultAll samples were examined using SPF10 primers for HPV detection. HPV DNA was positive in 8 out of 93 (8.6%) ESCC specimens. Using INNO-LiPA genotyping system we detected the genotypes of 5 out of 8 HPV-positive samples. Both HPV types 16 and 6 were detected in 3 specimens; one sample was positive for HPV type 18 and 2 samples were co-infected with two HPV types. There were no statistically significant differences between HPV-positive and HPV-negative cases with regard to clinical and pathologic findings. Three samples were positive for SPF10 indicating HPV infection; however, the exact HPV type could not be clarified using INNO-LiPA genotyping .ConclusionIn conclusion, the present study showed that a small proportion of ESCC specimens from Iran harbor HPV16, 18 genome using a highly sensitive method. As different rates have been reported from Iran, a more widespread study with more precise definition of geographical differences could delineate the potential involvement of HPV in the development of ESCC in Iranian population.     

Detection of human papillomavirus in Chinese esophageal squamous cell carcinoma and its adjacent normal epithelium

AIM: To investigate the putative role of human papillomavirus (HPV) infection in the carcinogenesis of esophageal squamous cell carcinoma in China. METHODS: Twenty-three esophageal squamous cell carcinoma samples and the distal normal epithelium from Shanxi Province, and 25 more esophageal squamous cell carcinoma samples from Anyang city, two areas with a high incidence of esophageal cancer in China, were detected for the existence of HPV-16 DNA by PCR, mRNA in situ hybridization (ISH) and immunohistochemistry (IHC) targeting HPV-16 E6 gene. RESULTS: There were approximately 64 % (31/48) patients having HPV-16 DNA in tumor samples, among them nearly two-thirds (19/31) samples were detected with mRNA expression of HPV-16 E6. However, in the normal esophageal epithelium from cancer patients, the DNA and mRNA of HPV-16 were found with much less rate: 34.7 % (8/23) and 26.1 % (6/23) respectively. In addition, at protein level detected by IHC assay, 27.1 % (13/48) tumor samples had virus oncoprotein E6 expression, while only one case of normal epithelium was found positive. CONCLUSION: HPV infection, especially type 16, should be considered as a risk factor for esophageal malignancies in China.     

Laryngeal squamous cell carcinoma in HIV-positive patients: lack of association with human papillomavirus infection

Objectives Neoplasms associated with human papillomavirus (HPV) infection occur at increased frequency in patients with HIV infection/AIDS. Although laryngeal squamous cell carcinomas (LSCCs) in HIV-positive patients are uncommon, a higher incidence of this malignancy in HIV-positive patients than in the general population has been reported. As a proportion of LSCCs are associated with HPV in the general population, the clinicopathological features of a series of LSCCs developing in HIV-positive patients were evaluated to investigate the possible relationship with HPV infection, and infection with other oncogenic viruses.
Methods All HIV-positive patients with LSCC diagnosed at a single institution from 1998 to 2007 were retrospectively evaluated. The clinicopathological features were analysed and tissues were tested by polymerase chain reaction (PCR), using the short PCR fragment 10 (SPF10) primer, a highly sensitive method for HPV DNA detection. Immunohistochemical studies for HIV p24, p16^INK4a and p53 were performed. Epstein–Barr virus (EBV) and human herpes virus 8 (HHV-8) were also investigated.
Results Six out of 4987 HIV-infected patients seen in this period in the Infectious Diseases Department developed LSCC (median age 41.5 years; male to female ratio 1:1). All patients were heavy smokers and the tumours presented at an advanced clinical stage. HPV was not detected in any tumour, not even in two patients with coexisting HPV-associated gynaecological neoplasm. Staining for HIV p24 and p16^INK4a was negative, whereas p53 was positive in half the cases. EBV and HHV-8 were also negative.
Conclusion LSCC developing in HIV-positive patients is an infrequent neoplasm, not usually associated with HPV infection. It develops in young, heavy smokers and presents at an advanced clinical stage.     

High prevalence of human papillomavirus in esophageal squamous cell carcinoma: a study in paired samples

Esophageal squamous cell carcinoma (ESCC) is one of the common cancers with a poor prognosis. Incidences of human papillomavirus (HPV) infection range from 0 to 67% in different parts of the world. It has been frequently associated with high‐risk HPV genotypes 16 and 18. The present study analyzes the prevalence of HPV infection in ESCC tumor and adjoining mucosa. Fresh tissue samples were obtained from ESCC tumor (group I) and adjoining mucosa (group II). Aliquots of DNA extracts were used. There were 23 patients with paired samples, 19 (83%) were male. HPV was positive in 20/23 (87%). Mean age of HPV positive in group I was 56.63 ± 6.96 and in group II 54.31 ± 7.13 years (P > 0.05). Majority had more than one viral type. HPV52 was the most common observed in 14 (61%) males and two (9%) females. Other common viruses were HPV55, 39, and 59. Smoking had a significant association with viral positivity. p63 and p16 oncoproteins correlated with degree of tumor differentiation but not with viral status. We documented high prevalence of high‐risk HPV in ESCC. Our observations support the concept of persistent infection by an oncogenic HPV in cancer development. Our study highlights importance of documenting viral genotype in a defined geographic area.     

Expression of CD44 variants and prognosis in oesophageal squamous cell carcinoma

BACKGROUND: The CD44 variant (CD44v) isoforms have been noted as markers for tumour metastasis and prognosis in several adenocarcinomas. AIMS: To investigate whether CD44v, especially the CD44v2 (v2) isoform, may be a useful prognostic factor for patients with oesophageal squamous cell carcinoma, using a recently developed monoclonal antibody against a v2 epitope. PATIENTS: 233 patients (211 men and 22 women; mean age 61.9 years), with oesophageal squamous cell carcinomas curatively removed without additional treatment between 1987 and 1996 at the National Cancer Center Hospital, were analysed for CD44v expression. METHODS: The expression of CD44v was evaluated immunohistochemically using monoclonal antibodies against epitopes of the standard and variant protein, in paraffin embedded oesophageal squamous cell carcinoma tissue from 233 patients who had undergone cervical, mediastinal, and abdominal lymphadenectomy (three field dissection) for oesophagectomy. The data were evaluated for any correlation with clinicopathological indices or prognosis. RESULTS: Although total CD44 and CD44v6 (v6) were respectively observed in 99% and 97% of the cancer specimens, the expression of v2 was only 30%. Patients whose tumours were v2 positive had a significantly better prognosis than those whose tumours were v2 negative (p = 0.031). Furthermore, in patients without lymph node metastasis, v2 positivity alone was a significant independent factor of prognosis (relative risk of death associated with v2 negativity, 4.7; p = 0.037) in multivariate analysis. CONCLUSIONS: These results indicate that v2 is a useful marker for clinical prognosis in patients with oesophageal squamous cell carcinoma. Particularly in patients without lymph node metastasis, v2 status may thus have implications for the use of adjuvant chemotherapy and/or radiotherapy in patients with oesophageal cancer at an early stage.     

Study of human papillomavirus infection in patients with anal squamous carcinoma

PURPOSE: The purpose of this study was to determine the incidence of human papillomavirus deoxyribonucleic acid (HPV DNA) in anal squamous carcinoma. METHODS: HPV DNA in situ hybridization for HPV Types 6, 11, 16, 18, 31, 33, and 35 was performed on the formalin-fixed, paraffinembedded tissue from 53 perianal and anal squamous carcinomas and 10 controls. RESULTS: HPV DNA sequences were identified in 18 of 53 anal squamous carcinomas (34 percent). All 10 controls were negative for HPV DNA. Of the 18 positive patients, 10 were perianal squamous carcinomas, and 8 were anal canal squamous carcinomas. Six of the perianal carcinomas were positive for HPV Types 6 and 11. The remaining four perianal carcinomas and all eight of the anal canal carcinomas were positive for HPV Types 16 and 18. CONCLUSION: HPV DNA sequences can be identified in anal squamous carcinomas. Anal squamous epithelium is another site where HPV infection may carry a risk for malignant transformation. One-third of anal squamous carcinomas may be associated with prior HPV infection. Patients with anogenital HPV infection should be routinely screened for anal squamous lesions.     

Human papillomavirus in esophageal squamous cell carcinoma in Colombia and Chile

INTRODUCTION Human papillomavirus (HPV), a double-stranded DNA virus, is recognized as an etiologic agent of cervical cancer[1]. In addition, HPV is suspected of causing extragenital cancers, including cancers of the oral cavity, larynx, esophagus, and lu…     

Human papillomavirus in squamous cell carcinoma of esophagus in a high-risk population

AIM: To investigate the relation of human papillomavirus (HPV) and esophageal squamous cell carcinoma (ESCC) in Iranian patients as compared to normal controls. METHODS: Using MY09/MY11 consensus primers, we compared the prevalence of a HPV L1 gene in tumor tissues from 38 ESCC cases and biopsied tissues from 38 endoscopically normal Iranian individuals. We also compared the presence of HPV16 and HPVA18 in the same samples using type-specific E6/E7 primers. RESULTS: Fourteen (36.8%) of the 38 ESCC samples but only 5 (13.2%) of the 38 control samples were positive for the HPV L1 gene (P= 0.02). Five (13.2%) of the ESCC samples but none of the control samples were positive for the HPV16 E6/E7 gene (P= 0.05). Three (7.9%) of the ESCC samples and 5 (13.2%) of the control samples were positive for the HPV18 E6/E7 gene (P= 0.71). CONCLUSION: Our data are consistent with HPV DNA studies conducted in other high-risk areas for ESCC. HPV should be considered as a potential factor contributing to the high incidence of ESCC in Iran and other high-incidence areas of the world.     

An investigation into the role of human papillomavirus in endobronchial papillary squamous tumours

Tissue specimens from 15 patients with endobronchial papillary squamous tumours were probed for the presence of human papillomavirus DNA using the technique of in situ hybridization with probes to human papillomavirus genotypes 4, 5, 6b, 8, 11, 16 and 18. Despite the histological similarity of these lesions to known human papillomavirus-associated tumours no evidence of hybridization was detected in any of the cases analysed. It is thus unlikely that human papillomavirus has a significant role in the genesis of these tumours.     

Association between esophageal squamous cell carcinoma and human papillomavirus detected by Hybrid Capture II assay

From July 1999 to July 2000, 40 patients from the Surgical Oncology and Gastroenterology Departments of Irmandade da Santa Casa de Misericórdia de Porto Alegre were enrolled in a prospective study in order to find the frequency of human papillomavirus DNA (HPV-DNA) in esophageal squamous cell carcinoma. They were compared to 10 normal individuals (controls). Digene Inc. (Digene Corporation, Gaithersburg MD, USA) Hybrid Capture II assay, a chemoluminescent method, was used in both groups. The samples were collected from the tumor group by endoscopic biopsy in 20 cases, and by surgical specimens in the other 20 cases; the control group all underwent endoscopic biopsies. The cut-off point in the analytical assay for HPV-DNA was 1.0 pg/mL. Digene Corp. Hybrid Capture II assay tests HPV-DNA and differentiates high and low-risk for malignization virus groups. Gender, age, location of the tumor in esophagus and degree of differentiation on histology were also analyzed. There was one positive case (1.14 pg/mL), a 2.5% frequency of low-risk HPV-DNA type in the esophageal squamous cell carcinoma group (CI 95%: 0.00-7.34%). This group of patients consisted of 30 male and 10 female individuals, similar to the distribution seen in large series of such patients from the Brazilian population. The mean age was 63 years. Endoscopic samples of 10 patients who underwent endoscopic biopsy procedure for aleatory reasons, with normal looking esophageal mucosa, were tested to detect HPV-DNA, as controls. There was one positive case (1.17 pg/mL) of high-risk HPV-DNA type, resulting in a 10% positive rate. HPV-DNA is rarely found in esophageal squamous cell carcinomas: we found a 2.5% frequency, with confidence interval of 0-7.34%; and this is in the same range of controls (10% frequency, with CI: 9.1-10.9%). Both patients had a viral load quite near the 1.0 pg/mL cut-off point.     

HPV16与新疆维吾尔族食管鳞癌的相关性

目的:调查新疆维吾尔族食管鳞癌患者中人乳头瘤状病毒(HPV)16型的感染率.方法:采用PCR技术检测82例新疆维吾尔族食管鳞癌和80例癌旁正常食管黏膜石蜡包埋组织HPV16的感染率,分析其与新疆维吾尔族食管鳞癌发生的关联性.结果:在新疆维吾尔族食管鳞癌组织中,HPV16型检出率为31.7%(26/82),与对照组12.5%(10/80)相比,差异有统计学意义(X2=8.643,P<0.05);按照年龄、性别、病理分化程度及临床分期对维吾尔族食管鳞癌中的HPV16感染状况比较,差异均无统计学意义(X2=0.301、0.149、2.876、0.105,均P>0.05).结论:HPV16型感染与新疆维吾尔族食管鳞癌的发生存在一定的相关性,但与患者的年龄、性别、病理分化程度及临床分期无明显的关联性.