Skip Navigation

This Article
Right arrow Abstract Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (15)
Right arrow Request Permissions
Google Scholar
Right arrow Articles by Mizobuchi, S
Right arrow Articles by Terada, M
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mizobuchi, S
Right arrow Articles by Terada, M
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Japanese Journal of Clinical Oncology Pages 1-5

Absence of Human Papillomavirus-16 and -18 DNA and Epstein-Barr Virus DNA in Esophageal Squamous Cell Carcinoma
Introduction
Materials and Methods
   Extraction of DNA from surgical specimens and cultured cell lines
   PCR analysis and Southern blot hybridization for HPV-16 and HPV-18 DNA
   Sensitivity of PCR for amplifying HPV-16 and HPV-18 DNA
   PCR analysis for EBV DNA
Results
   Detection of HPV DNAs
   Detection of EBV DNA
Discussion
Acknowledgements
References

Absence of Human Papillomavirus-16 and -18 DNA and Epstein-Barr Virus DNA in Esophageal Squamous Cell Carcinoma

Absence of Human Papillomavirus-16 and -18 DNA and Epstein-Barr Virus DNA in Esophageal Squamous Cell Carcinoma Shunji Mizobuchi1, Hiromi Sakamoto1, Yuji Tachimori2, Hoichi Kato2, Hiroshi Watanabe2 and Masaaki Terada1

1Genetics Division, National Cancer Center Research Institute, 2Department of Surgery, National Cancer Center Hospital, Tokyo, Japan

To elucidate the association of human papillomavirus (HPV) and Epstein-Barr virus (EBV) with carcinogenesis of the esophagus, 41 surgically resected specimens and 12 cell lines of esophageal squamous cell carcinoma were examined for the presence of HPV DNA and EBV DNA by polymerase chain reaction using primers for the E6 regions of HPV-16 and -18 and for the EBNA 1 region of EBV. We designed the reaction condition to amplify HPV and EBV DNA specifically and detected by gel electrophoresis. In ethidium bromide staining, no band was detected either for the HPV E6 region or for the EBV EBNA 1 gene in any of the surgically resected specimens and the cell lines, although the HPV sequence was detectable by Southern blot hybridization, which is a more sensitive detection method than staining; three out of 41 surgically resected specimens were positive for HPV-18 by Southern blot hybridization of polymerase chain reaction products. However, the number of viral genomes has been estimated as lower than 1 X 10-3 copies per cell based on the intensity of the hybridization signals. Moreover, the DNA samples extracted from the corresponding non-cancerous esophageal mucosa were also positive for HPV-18, and the intensities of the hybridization signals were almost the same as those of the tumors. The results of our study indicate that HPV-16, HPV-18 and EBV are not generally associated with esophageal carcinogenesis.

Key words: human papillomavirus - Epstein-Barr virus - polymerase chain reaction - esophageal squamous cell carcinoma - esophageal carcinogenesis

Introduction

The recent interest in human papillomavirus (HPV) and its association with cervical squamous cell carcinoma (SCC) (1-4) has prompted the investigation for its possible etiological role in cancers at other body sites. In lesions of the skin (5), oral cavity (6), tongue (7), larynx (8) and lung (9), the presence of HPV was investigated initially by DNA hybridization techniques and recently by polymerase chain reaction (PCR).

HPV infection of the esophagus was first suggested by morphologic studies showing HPV-induced cytopathic changes (10) and later by immunohistochemical techniques demonstrating the presence of HPV antigens (11,12). The presence of the HPV genome in SCC of the esophagus was demonstrated by DNA hybridization techniques and/or PCR. However, the HPV detection rates vary considerably among different authors, ranging from 0% to 49% (13-22). In Japan, Furihata et al. (13) reported that 33.8% of patients with esophageal SCC were positive for HPV DNA by in situ hybridization and Toh et al. (14) reported a prevalence of 6.7% by PCR. It is important to establish whether HPV DNA is demonstrated in Japanese esophageal carcinoma and whether this virus plays a role in the etiology of the disease. In this study we used the PCR technique to detect HPV-16 and HPV-18 DNAs with primers amplifying a portion of the E6 region. The specific HPV DNA sequences amplified by PCR were subsequently confirmed by Southern blot hybridization with the HPV-specific probes.

The Epstein-Barr virus (EBV) was first identified in 1964 in cultured Burkitt's lymphoma cells (23) and has since been implicated in a variety of benign and malignant lymphoproliferative diseases. The association of EBV with undifferentiated nasopharyngeal carcinoma has been well documented (24,25). However, it remains controversial whether the nasopharyngeal SCC are also EBV-associated. Recently, the involvement of EBV has been demonstrated in gastric adenocarcinoma of various grades of differentiation without prominent lymphoid infiltration (26, 27). Thus it is of interest to determine whether EBV is involved in carcinoma of the esophagus, which is located between the nasopharynx and stomach. In this study we investigated the presence of EBV in squamous cell carcinoma of the esophagus by PCR.

Materials and Methods

Extraction of DNA from surgical specimens and cultured cell lines

The surgically resected specimens of esophageal carcinoma were obtained from the National Cancer Center Hospital at the time of surgery, snap-frozen in liquid nitrogen immediately after resection, and stored at -80°C until DNA extraction. Human esophageal carcinoma cell lines TE1-TE6 and TE8-TE13 were kindly provided by Dr. T. Nishihira (Second Department of Surgery, Faculty of Medicine, Tohoku University). Each cell line was established from SCC and was maintained in RPMI-1640 with 7% fetal bovine serum. Human cervical cancer cell lines SKG-IIIa and HeLa were maintained in Ham's F12 medium with 10% fetal bovine serum and in Eagle's minimum essential medium with 10% calf serum respectively. SKG-IIIa cells and HeLa cells contain approximately one copy of HPV-16 DNA and 10-20 copies of HPV-18 sequence per haploid genome respectively (2).

DNA was extracted as previously described (28) from the surgical specimens and cell lines. All the DNA samples were suitable for PCR analysis, because the 170 bp (base pair) fragment of the EXP1 gene, which is a gene identified on chromosome 11q13, about 120kb apart from the HST1 gene (29), was successfully amplified (data not shown). The EXP1 primers are:P40, 5'-AGCATTTGTGCCTGAAGCTGC-3' andP9, 5'-TGAAACCTCTCATCCGCCATCACGA-3'.

PCR reaction was performed in a 50 µl volume with 400 ng of template DNAs, 1x PCR Buffer, 10 mM of each deoxynucleotide triphosphate, 0.5 µg of each primer and 2.5 units of Taq-polymerase. The conditions for PCR analyses were as follows: 94°C for 10 min (1 cycle); 94°C for 1 min, 60°C for 2 min, 72°C for 3 min (30 cycles); 72°C for 10 min (1 cycle). After amplification of PCR, 10 µl (corresponding to 1/5 volume of the reaction mixture) was subjected to gel electrophoresis on a 3.5% NuSieve 3:1 agarose gel and stained with ethidium bromide. In ethidium bromide staining, the amplified 170 bp fragments corresponding to a DNA fragment of EXP1 used as an internal control were seen in all of the surgical specimens and the cell lines (data not shown), confirming that all DNA samples were suitable for PCR analysis.

PCR analysis and Southern blot hybridization for HPV-16 and HPV-18 DNA

Oligonucleotide primers were synthesized by a 392 DNA/RNA synthesizer (Perkin Elmer Co., Connecticut) according to published sequences (30). Sequences of primers for amplification of the E6 regions of HPV-16 and HPV-18 are as follows:p16F, 5'-AAGGGCGTAACCGAAATCGGT-3';

p16R, 5'-GTTTGCAGCTCTGTGCATA-3';p18F, 5'-AAGGGAGTAACCGAAAACGGT-3';p18R, 5'-GTGTTCAGTTCCGTGCACA-3'.

p16F and p18F correspond to the E6 sense sequence of HPV-16 and HPV-18 respectively. p16R and p18R correspond to the E6 antisense sequence of HPV-16 and HPV-18 respectively.

Each PCR reaction mixture (100 µl) contained 600 ng of template DNA, 1x PCR Buffer (10 mM Tris-HCl, pH 8.3, 50 mM KCl, 1.5 mM MgCl2, 0.001% (W/V) gelatin), 10 mM of each deoxynucleotide triphosphate, 0.15 µg of each primer and 2.0 units of Taq-polymerase. The PCR-cycling reactions were performed on a Perkin-Elmer Cetus (Norwalk, CT) DNA thermal cycler. After one cycle of denaturation at 94°C for 10 min, 30 cycles of 94°C for 1 min, 55°C for 2 min and 72°C for 2 min were carried out followed by extension at 72°C for 10 min. All experiments were performed in parallel with positive controls (SKG-IIIa for HPV-16 and HeLa for HPV-18) and negative controls (HeLa for HPV-16 and SKG-IIIa for HPV-18).

After amplification of PCR, 10 µl (1/10 volume) of the reaction mixture was subjected to gel electrophoresis on a 3.5% NuSieve 3:1 agarose gel and stained with ethidium bromide. DNA was transferred to the Hybond-N (Amersham International plc, Buckinghamshire, UK) membrane filter. Hybridization was performed at 42°C for 8-12 h in a hybridization solution (50% formamide, 0.1 M PIPES-NaOH, pH 6.8, 0.65 M NaCl, 5x Denhardt's solution, 5 mM EDTA, 0.1% SDS, 10% Dextran sulfate, 100 µg/ml denatured salmon testis DNA and 32P-labeled HPV-16 or HPV-18 probe at 1*106 c.p.m./ml). After hybridization, the filters were washed and exposed at -80°C to Kodak XAR-5 film with an intensifying screen for 1 to 4 days.

Sensitivity of PCR for amplifying HPV-16 and HPV-18 DNA

To define the sensitivity of the PCR, DNA from SKG-IIIa cells or HeLa cells was serially diluted with salmon testis DNA and was subjected to PCR analysis for HPV-16 or HPV-18. After amplification by PCR, the reaction mixture was electrophoresed, blotted and hybridized with 32P-labeled HPV-16 or HPV-18 DNA as a probe .

PCR analysis for EBV DNA

Sequences of EBV primers, from published data (31) are as follows:SL1, 5'-GGACCTCAAAGAAGAGGGGG-3',SL3, 5'-GCTCCTGGTCTTCCGCCTCC-3'.

The primers amplify and detect an 80-base pair region of the EBNA1 gene. Each PCR reaction mixture (100 µl) contained 600 ng of template DNAs, 1x PCR Buffer, 10 mM of each deoxynucleotide triphosphate, 0.25 µg of each primer, 1.0 unit of Perfect Match (Stratagene, California) and 3.0 units of Taq-polymerase. The conditions for PCR analyses were as follows: 94°C for 10 min (1 cycle); 95°C for 45 sec, 60°C for 45 sec, 72°C for 60 sec (35 cycles); 72°C for 10 min (1 cycle). All experiments were performed in parallel with pDR 2 (Clontech Laboratories, Inc., California), which is the vector containing the EBV EBNA 1 sequence, as a positive control, and with a negative control in which PCR reaction was performed without a template DNA. After PCR amplification, 10 µl of the reaction mixture was subjected to gel electrophoresis on a 3.5% NuSieve 3:1 agarose gel and stained with ethidium bromide.

Results

Detection of HPV DNAs

The PCR products amplified with the two sets of primers specific for HPV-16 or HPV-18 DNAs were observed as a specific band of the expected size (140 bp) in the positive controls, SKG-IIIa for HPV-16 and HeLa for HPV-18 (Fig. 1). No band was detected in DNAs from HeLa and SKG-IIIa cells which were used as negative controls for HPV-16 and HPV-18 DNA, respectively.


Figure 1. Southern blot hybridization analysis of PCR products for the presence of HPV-18 DNA. Lanes 1, 2 and 3: HeLa cell DNA, containing 10 to 20 copies HPV-18 DNA sequence per haploid genome, was serially diluted with salmon testis DNA. Lane 1, dilution at 1/50; lane 2, 1/5 000; lane 3, 1/50 000. Lanes 4-13, five pairs of the cancerous portions (even numbers) and non-cancerous portions (odd numbers) of the surgical specimens of the esophagus. The band hybridizing to the HPV-18 probe was visible on lane 4 upon an extended exposure. Lane 14: SKG-IIIa cell DNA, containing HPV-16 DNA but not HPV-18 DNA.

Ethidium bromide staining of the gel revealed no detectable band for any of the 41 surgical specimens or the 12 cell lines. Moreover, by Southern blot hybridization of PCR products, no HPV-16 genome was detectable in any of the surgical specimens or the cell lines (data not shown). Although all of the 12 cell lines were negative, three out of 41 surgical specimens were positive for HPV-18 DNA. The intensity of their hybridization signals was weaker than that of 1/50 000 diluted HeLa cell DNA. In the DNA samples extracted from the corresponding non-cancerous esophageal mucosae of the same patients, an HPV-18 DNA fragment was detected with almost the same intensity of hybridization signals as that of the tumor counterparts (Fig. 1).

Comparison of the intensities of the serially diluted DNA fragments demonstrates that the method we used could detect, in PCR products, 0.1 copies per haploid genome by ethidium bromide staining and 10-3 copies per haploid genome by Southern blot hybridization, of both HPV-16 and HPV-18 DNAs (Fig. 2 and data not shown).


Figure 2. Sensitivity of detection of HPV DNA. SKG-IIIa DNA (one copy of HPV-16 DNA sequence per haploid genome) was diluted serially with salmon testis DNA. Number indicates number of copies of HPV DNA in the diluted DNA specimens. 10-3 copies of HPV-16 sequence could be detected, while 10-4 copies could not.

Detection of EBV DNA

A specific band of the expected size of 80 bp should be detected in the PCR products amplified with two primers specific for EBV DNA, if EBV EBNA 1 gene is present in a DNA sample. No EBV EBNA1 specific DNA could be detected by ethidium bromide staining in any of the 41 surgical specimens or the 12 cell lines (Fig. 3).

Discussion

Kulski et al. (32) detected HPV DNA in esophageal SCC (four of 10 cases) in Western Australia by filter in situ hybridization. However, Loke et al. (15) were unable to detect HPV genome in 37 cases of esophageal SCC in Hong Kong by slot blot hybridization or in situ hybridization. In France, one of 12 esophageal SCC was positive for HPV-18 DNA by in situ hybridization and three others were positive by dot blot hybridization (16). In the Linxian district of China, the high-incidence area for esophageal carcinoma, 71 (19.6%) of 363 patients with esophageal SCC were shown to have HPV DNA in their cancer cells by in situ hybridization (17). In Japan, 24 (33.8%) of 71 patients with esophageal SCC were positive for HPV DNA by in situ hybridization (13). Of the 24 cases, 10 were HPV-16 and the others were HPV-18.


Figure 3. Absence of EBV genome in esophageal cancer. All the DNA samples were subjected to PCR for amplification of 80 bp, corresponding to a portion of EBNA 1 gene of EBV, as described in the text. Lane 1: pDR 2 DNA, which is the plasmid containing the EBV EBNA 1 sequence, as a positive control. Lane 2: no template DNA, as a negative control. Lane 3-6: Absence of EBV EBNA 1 specific DNA in esophageal cancers. All the other DNA samples tested were also negative for EBV DNA. Lane M: DNA size marker, [Phi]x174 digested by HaeIII.

Among the PCR-based studies for the HPV-DNA detection, 25 out of 51 (49.0%) biopsy specimens in esophageal precancerous lesions and squamous cell carcinomas have been reported as being positive in the Linxian district of China (33). The specific HPV DNA sequences amplified by PCR were subsequently confirmed by Southern blot hybridization with specific HPV probes. Togawa et al. (18) identified HPV DNA in 14% (10/72) of esophageal squamous cell carcinomas from different regions of the world by a radioactive nested PCR /restriction fragment length polymorphism analysis. By contrast, Kiyabu et al. (19) detected neither HPV-16 nor HPV-18 DNA in 13 esophageal carcinomas in North American patients, using PCR with primers which amplify the E6 of both HPV-16 and HPV-18. Smits et al. (20) detected HPV DNA in none of the 61 esophageal squamous cell carcinomas by multiple broad spectrum PCR. Among 14 patients with esophageal SCC in South Africa, 6 (42.9%) were positive for HPV DNA in tumor biopsies by PCR with primers which amplify the L1 gene of a wide range of HPVs (21). Toh et al. (14) in Japan showed by PCR that three (6.7%) of 45 biopsy samples of esophageal SCC contained HPV-16 or HPV-18 DNA. In this study, primers for PCR were firstly consensus primers for the simultaneous amplification of the E6-E7 regions for cancer-associated HPV types and secondarily type-specific primers for the E7 regions. In the latter three reports, confirmation by Southern blot hybridization of PCR products has not been performed.

Marked variations in the prevalence of esophageal HPV infections have been reported by different authors. This difference might be due to the difference in the methods used. We have used PCR analysis coupled with Southern blot hybridization to detect the E6 region of the HPV-16 or HPV-18 genome, with adequate positive and negative control. The sensitivity of our PCR detection system of the HPV genome was 0.1 copies per haploid genome by ethidium bromide staining and 10-3 copies per haploid genome by Southern blot hybridization of PCR products.

In the present series, three out of 41 surgical specimens were positive for HPV-18 DNA by Southern blot hybridization of PCR products. However, the intensity of hybridization signals in these cases was weaker than that of 1/50 000 diluted HeLa cell DNA, which carries 10-50 integrated copies of HPV-18 DNA per haploid genome (2), indicating that the number of viral genomes in those samples is probably lower than 1*10-3 copies per haploid genome. The possibility of the specific presence of the HPV genome in a clonal population of the tumor cells is quite unlikely. Moreover, HPV-18 was also positive in DNA extracted from their normal esophageal epithelium and the intensity of the hybridization signals was almost the same as that of the tumor cell. Therefore we presume that HPV is present in fewer than one in 103 cells in the cancerous as well as non-cancerous portion of the esophagus of those patients as a fortuitous parasite of the organ.

While there is a report indicating 33.8% positive for HPV DNA by in situ hybridization in Japan (13), this has not been confirmed in other studies. The reason for the discrepancy remains to be elucidated, but it could be due to a difference in the methods employed. It is also possible that there may be a geographic variation in the prevalence of HPV infection in Japan.

Recently, an association of EBV with epithelial malignancies without prominent lymphoid infiltration has been demonstrated, especially in gastric adenocarcinoma (26). Very little information is available yet in the literature with respect to the possible association of EBV with another upper digestive tract cancer, SCC of the esophagus. Niedobitek et al. (25) have demonstrated the absence of detectable EBV-DNA in any of the eight squamous cell NPCs, 26 SCCs of the tonsils and 14 cervical cancers examined. We have demonstrated the absence of detectable EBV-DNA in all of the 41 surgical specimens and 12 cell lines of the SCCs of the esophagus.

Our PCR analyses detected no specific association of HPV or EBV genome with esophageal SCCs. The present results strongly suggest that HPV-16, HPV-18 and EBV do not play any significant role in the development of esophageal carcinomas.

Acknowledgements

We thank Dr. T. Nishihira (Second Department of Surgery, Faculty of Medicine, Tohoku University) for the gifts of the cell lines, Drs. Y. Matsusima, T. Fujii (Department of Obstetrics and Gynecology, School of Medicine, Keio University) and H. Tsuruta (First Department of Surgery, Nippon Medical School) for helpful discussions. This study was supported in part by a Grant-in-Aid for the Second Term Comprehensive 10-year Strategy for Cancer Control from the Ministry of Health and Welfare and by Grants-in-Aid for Cancer Research from the Ministry of Health and Welfare in Japan.

References

1. Dürst M, Gissmann L, Ikenberg H, zur Hausen H. A papillomavirus DNA from a cervical carcinoma and its prevalence in cancer biopsy specimens from different geographic regions. Proc Natl Acad Sci USA 1983; 80:3812-5. MEDLINE Abstract

2. Tsunokawa Y, Takebe N, Nozawa S, Kasamatsu T, Gissmann L, zur Hausen H et al. Presence of human papaillomavirus type-16 and type-18 DNA sequences and their expression in cervical cancers and cell lines from Japanese patients. Int J Cancer 1986;37:499-503. MEDLINE Abstract

3. zur Hausen H, de Villiers EM. Human papillomaviruses. Ann Rev Microbiol 1994;48:427-447. MEDLINE Abstract

4. Palefsky JM, Holly E.A. Molecular virology and epidemiology of human papillomavirus and cervical cancer. Cancer Epidemiology, Biomarkers & Prevention 1995;4:415-28. MEDLINE Abstract

5. Grimmel M, de Villiers EM, Pawlita M, Neumann C, zur Hausen H. Characterization of a new human papillomavirus type (HPV 41) isolated from disseminated warts and the detection of closely related sequences in some squamous cell carcinomas. Int J Cancer 1988;45:5-9.

6. Maitland NJ, Bromidge T, Cox MF, Crane IJ, Prime SS, Scully C. Detection of human papillomavirus genes in human oral tissue biopsies and cultures by polymerase chain reaction. Br J Cancer 1989;59:698-703. MEDLINE Abstract

7. de Villiers EM, Weidauer H, Otto H, zur Hausen H. Papillomavirus DNA in human tongue carcinomas. Int J Cancer 1985;36:575-8. MEDLINE Abstract

8. Scheurlen W, Stremlau A, Gissmann L, Horn D, Zenner HP, zur Hausen H. Rearranged HPV-16 molecules in an anal carcinoma and in a laryngeal carcinoma. Int J Cancer 1986;38:671-6. MEDLINE Abstract

9. Syrjänen K, Syrjänen S, Kellokoski J, Kärjä J, Mäntyjärvi R. Human papillomavirus (19HPV) type 6 and 16 DNA sequences in bronchial squamous cell carcinomas demonstrated by in situ DNA hybridization. Lung 1989;167:33-42. MEDLINE Abstract

10. Syrjänen KJ. Histological changes identical to those of condylomatous lesions found in esophageal squamous-cell carcinomas. Arch Geschwulstforsch 1982;52:283-92. MEDLINE Abstract

11. Winkler B, Capo V, Reumann W, Ma A, La Porta R, Reilly S et al. Human papillomavirus infection of the esophagus. Cancer 1985;55:149-55. MEDLINE Abstract

12. Mori M, Shimono R, Inoue T, Kuwano H, Sugimachi K, Zhang RG. Papillomavirus and esophageal cancer in the Japanese and Chinese. Am J Gastroenterol 1989;84:1126-7. MEDLINE Abstract

13. Furihata M, Ohtsuki Y, Ogoshi S, Takahashi A, Tamiya T, Ogata T. Prognostic significance of human papillomavirus genomes (19type-16, -18) and aberrant expression of p53 protein in human esophageal cancer. Int J Cancer 1993;54:226-30. MEDLINE Abstract

14. Toh Y, Kuwano H, Tanaka S, Baba K, Matsuda H, Sugimachi K et al. Detection of human papillomavirus DNA in esophageal carcinoma in Japan by polymerase chain reaction. Cancer 1992;70:2234-8. MEDLINE Abstract

15. Loke SL, Ma L, Wong M, Srivastava G, Lo I, Bird CC. Human papillomavirus in oesophageal squamous cell carcinoma. J Clin Pathol 1990;43:909-12. MEDLINE Abstract

16. Benamouzig R, Pigot F, Quiroga G, Validire P, Chaussade S, Catalan F et al. Human papillomavirus infection in esophageal squamous-cell carcinoma in Western countries. Int J Cancer 1992;50:549-52. MEDLINE Abstract

17. Chang F, Syrjänen S, Shen Q, Wang L, Syrjänen K. Screening for human papillomavirus infections in esophageal squamous cell carcinomas by in situ hybridization. Cancer 1993;72:2525-30. MEDLINE Abstract

18. Togawa K, Jaskiewicz K, Takahasi H, Meltzer SJ, Rustgi AK. Human papillomavirus DNA sequences in esophagus squamous cell carcinoma. Gastroenterology 1994;107:128-36.

19. Kiyabu MT, Shibata D, Arnheim N, Martin WJ, Fitzgibbons PL. Detection of human papillomavirus in formalin-fixed, invasive squamous carcinomas using the polymerase chain reaction. Am J Surg Pathol 1989;13:221-4. MEDLINE Abstract

20. Smits HL, Tjong-A-Hung SP, ter Schegget J, Nooter K, Kok T. Absence of human papillomavirus DNA from esophageal carcinoma as determined by multiple broad spectrum polymerase chain reactions. J Med Virol 1995;46: 213-15. MEDLINE Abstract

21. Williamson AL, Jaskiesicz K, Gunning A. The detection of human papillomavirus in oesophageal lesions. Anticancer Res 1991;11:263-6. MEDLINE Abstract

22. Kulski J, Demter T, Sterrett, GF, Shilkin KB. Human papilloma virus DNA in oesophageal carcinoma. Lancet 1986;II:683-4. MEDLINE Abstract

23. Epstein MA, Achong BG, Barr YM. Virus particles in cultured lymphoblasts from Burkitt's lymphoma. Lancet 1964;I:702-3.

24. Wolf H, zur Hausen H, Becker V. EB viral genomes in epithelial nasopharyngeal carcinoma cells. Nature 1973;44:245-7.

25. Niedobitek G, Hansmann ML, Herbst H, Young LS, Dienemann D, Hartmann CA et al. Epstein-Barr virus and carcinomas: Undifferrentiated carcinomas but not squamous cell carcinomas of the nasopharynx are regularly associated with the virus. J Pathol 1991;165:17-24. MEDLINE Abstract

26. Shibata D, Weiss LM. Epstein-Barr virus-associated gastric adenocarcinoma. Am J Pathol 1992;140:769-74. MEDLINE Abstract

27. Tokunaga M, Uemura Y, TokudomeT, Ishidate T, Masuda H, Okazaki E et al. Epstein-Barr virus related gastric cancer in Japan: A molecular patho-epidemiological study. Acta Pathol Jpn 1993;43:574-81. MEDLINE Abstract

28. Sakamoto H, Mori M, Taira M, Yoshida T, Matsukawa S, Shimizu K et al. Transforming gene from human stomach cancers and a noncancerous portion of stomach mucosa. Proc Natl Acad Sci USA 1986;83:3997-4001. MEDLINE Abstract

29. Akiyama N, Tsuruta H, Sasaki H, Sakamoto H, Hamaguchi M, Ohmura Y et al. Messenger RNA levels of five genes located at chromosome 11q13 in B-cell tumors with chromosome translocation t(11;14)(19q13;q32). Cancer Res 1994;54:377-9. MEDLINE Abstract

30. Shimada M, Fukushima M, Mukai H, Kato I, Nishikawa A, Fujinaga K. Amplification and specific detection of transforming gene region of human papillomavirus 16, 18 and 33 in cervical carcinoma by means of the polymerase chain reaction. Jpn J Cancer Res 1990;81:1-5. MEDLINE Abstract

31. Shibata D, Weiss LM, Nathwani BN, Brynes RK, Levine AM. Epstein-Barr virus in benign lymph node biopsies from individuals infected with the human immunodeficiency virus is associated with concurrent or subsequent development of non-Hodgkin's lymphoma. Blood 1991;77:1527-33. MEDLINE Abstract

32. Kulski J, Demter T, Sterrett, GF, Shilkin KB. Human papilloma virus DNA in oesophageal carcinoma. Lancet 1986;II:683-684. MEDLINE Abstract

33. Chang F, Syrjänen S, Shen Q, Wang L, Wang D, Syrjänen K. Human papillomavirus involvement in esophageal precanerous lesions and squamous cell carcinoma as evidenced by microscopy and different DNA techniques. Scand J Gastroenterol 1992;27:553-63. MEDLINE Abstract

Received April 22, 1996; accepted July 15, 1996
For reprints and all correspondence: Masaaki Terada, National Cancer Center Research Institute, 1-1, Tsukiji 5-chome, Chuo-ku, Tokyo, 104, Japan
Abbreviations: HPV, human papillomavirus; EBV, Epstein-Barr virus; PCR, polymerase chain reaction; SCC, squamous cell carcinoma

This page is run by Oxford University Press, Great Clarendon Street, Oxford OX2 6DP, as part of the OUP Journals
Comments and feedback: www-admin{at}oup.co.uk
Last modification: 19 May 1998
Copyright© Japanese Journal of Clinical Oncology, 1997.
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 J. Clin. Pathol.Home page
K J Syrjanen
HPV infections and oesophageal cancer
J. Clin. Pathol., October 1, 2002; 55(10): 721 - 728.
[Abstract] [Full Text] [PDF]

Home page
 CarcinogenesisHome page
T. Li, Z.-M. Lu, K.-N. Chen, M. Guo, H.-P. Xing, Q. Mei, H.-H. Yang, J. F. Lechner, and Y. Ke
Human papillomavirus type 16 is an important infectious factor in the high incidence of esophageal cancer in Anyang area of China
Carcinogenesis, June 1, 2001; 22(6): 929 - 934.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Epidemiol. Biomarkers Prev.Home page
J. Wang, A. Noffsinger, G. Stemmermann, and C. Fenoglio-Preiser
Esophageal Squamous Cell Carcinomas Arising in Patients from a High-Risk Area of North China Lack An Association with Epstein-Barr Virus
Cancer Epidemiol. Biomarkers Prev., December 1, 1999; 8(12): 1111 - 1114.
[Abstract] [Full Text]

This Article
Right arrow Abstract Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (15)
Right arrow Request Permissions
Google Scholar
Right arrow Articles by Mizobuchi, S
Right arrow Articles by Terada, M
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mizobuchi, S
Right arrow Articles by Terada, M
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?