Genome of Helicobacter pylori and Serotype of HPV Detected in Oropharyngeal and Laryngeal Cancer and Chronic Inflammation Patients
Abstract
:1. Introduction
- (1)
- To analyze of incidence of infection by HPV and HP in oropharyngeal and laryngeal cancer patients. Can we compare the incidence for cancer patients and chronic tonsillar inflammation to healthy persons?
- (2)
- Analysis of the HPV serotypes present in oropharyngeal and laryngeal cancer patients. Genomic analysis of Helicobacter pylori presents in oropharyngeal and laryngeal cancer patients. Determining the incidence of cytotoxin-associated protein (CagA), vacuolating cytotoxin (VacA), and the HPV subtypes in the cancer cohort and chronic inflammation group compared to healthy persons.
2. Materials and Methods
- Child and adolescent patients were excluded (age under 18).
- Pregnant women were excluded.
- The patients with any of the following: human immunodeficiency virus (HIV), Hepatitis B or C viruses.
- Any chronic Salmonella infections, tuberculosis, or zoonosis were excluded.
- The patients of any other cancer in history were excluded.
- The patients with any treatment of the immune system, such as HIV (incl. transplantation of any organs, among others), and patients on immunosuppression therapy (incl. steroids) were excluded.
- The patients suffering from chronic gastric inflammation and/or ulcers. The persons with a history of from reflux (gastro-esophageal reflux) were excluded.
- Antibiotic treatment administered within six months preceding the surgery or sample collecting.
- (a)
- Tissue of cancer by patients undergoing surgical treatment of oropharyngeal SCC.
- (b)
- Tissue of tonsils by patients undergoing surgical treatment of chronic tonsillitis, suffering one infection yearly for more than 5 years, or 2 infections in the last 3–5 years, or 4–5 infections in the last two years.
- (c)
- For the control group, we used the saliva samples from patients coming for a routine checkup appointment.
2.1. Collection and Handing of Samples
2.2. Detection Techniques
2.2.1. Real-Time PCR Amplification and Genotyping of Helicobacter pylori
2.2.2. Helicobacter pylori Confirmation by Commercial Real-Time PCR Assay
2.2.3. Human Papilloma Virus DNA Detection
2.2.4. Statistical Analyses
3. Results
4. Discussion
5. Conclusions
- The majority of HPV-infected patients has an HP infection at the same time in the same sample.
- The presence of HPV in oropharyngeal cancer patients alone is rare.
- Data in the study suggest that HP can influence local immune cells and may cause a decrease in immunity towards HPV and/or any other infectious agents.
- The CagA-positive HP was detected in less cases. This result of the study does not support the idea that CagA-positive HP is a primary carcinogen in the oropharyngeal area.
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Conflicts of Interest
References
- Genç, R.; Çağlı, S.; Yüce, I.; Vural, A.; Okuducu, H.; Patıroğlu, T.; Güney, E. The Role ofH. pyloriin the Development of Laryngeal Squamous Cell Carcinoma. Dis. Markers 2013, 35, 447–449. [Google Scholar] [CrossRef] [Green Version]
- Karczewska, E.; Konturek, J.E.; Konturek, P.C.; Cześnikiewicz, M.; Sito, E.; Bielański, W.; Kwiecień, N.; Obtułowicz, W.; Ziemniak, W.; Majka, J.; et al. Oral cavity as a potential source of gastric reinfection by Helicobacter pylori. Dig. Dis. Sci. 2002, 47, 978–986. [Google Scholar] [CrossRef]
- Burduk, P.K. Association between infection of virulence cagA gene Helicobacter pylori and laryngeal squamous cell carcinoma. Med. Sci. Monit. 2013, 19, 584–591. [Google Scholar] [CrossRef] [Green Version]
- Sivars, L.; Tani, E.; Näsman, A.; Ramqvist, T.; Munck-Wikland, E.; Dalianis, T. Human Papillomavirus as a Diagnostic and Prognostic Tool in Cancer of Unknown Primary in the Head and Neck Region. Anticancer. Res. 2016, 36, 487–493. [Google Scholar]
- Ramqvist, T.; Mints, M.; Tertipis, N.; Näsman, A.; Romanitan, M.; Dalianis, T. Studies on human papillomavirus (HPV) 16 E2, E5 and E7 mRNA in HPV-positive tonsillar and base of tongue cancer in relation to clinical outcome and immunological parameters. Oral. Oncol. 2015, 51, 1126–1131. [Google Scholar] [CrossRef] [Green Version]
- Lim, Y.; Totsika, M.; Morrison, M.; Punyadeera, C. Oral Microbiome: A New Biomarker Reservoir for Oral and Oropharyngeal Cancers. Theranostics 2017, 7, 4313–4321. [Google Scholar] [CrossRef]
- Bulut, Y.; Agacayak, A.; Karlidag, T.; Toraman, Z.A.; Yilmaz, M. Association of cagA+ Helicobacter pylori with Adenotonsillar Hypertrophy. Tohoku J. Exp. Med. 2006, 209, 229–233. [Google Scholar] [CrossRef] [Green Version]
- Pavlík, E.; Lukeš, P.; Potužníková, B.; Astl, J.; Hrdá, P.; Souček, A.; Matucha, P.; Doseděl, J.; Šterzl, I. Helicobacter pylori isolated from patients with tonsillar cancer or tonsillitis chronica could be of different genotype compared to isolates from gastrointestinal tract. Folia. Microbiol. 2007, 52, 91–94. [Google Scholar] [CrossRef]
- Abdel-Monem, M.H.; Magdy, E.; Nour, Y.A.; Harfoush, R.A.; Ibreak, A. Detection of Helicobacter pylori in adenotonsillar tissue of children with chronic adenotonsillitis using rapid urease test, PCR and blood serology: A prospective study. Int. J. Pediatric Otorhinolaryngol. 2011, 75, 568–572. [Google Scholar] [CrossRef]
- Lukes, P.; Astl, J.; Pavlík, E.; Potuzníková, B.; Sterzl, I.; Betka, J. Helicobacter pylori in tonsillar and adenoid tissue and its possible role in oropharyngeal carcinogenesis. Folia. Biol. 2008, 54, 33–39. [Google Scholar]
- Chen, M.; Fang, Y.; Cheng, L.; Wu, H. Helicobacter pylori is associated with poor prognosis of laryngeal precancerous lesion. Auris. Nasus. Larynx. 2020, 47, 268–275. [Google Scholar] [CrossRef]
- Pandey, S.; Follin-Arbelet, B.; Pun, C.B.; Gautam, D.K.; Johannessen, A.C.; Petersen, F.C.; Costea, D.E.; Sapkota, D. Helicobacter pylori was not detected in oral squamous cell carcinomas from cohorts of Norwegian and Nepalese patients. Sci. Rep. 2020, 10, 8737. [Google Scholar] [CrossRef]
- Fellmann, J.; Weisert, J.U.; Soltermann, A.; Morand, G.; Morra, L.; Moch, H.; Huber, G.F.; Probst, R. Helicobacter pylori detected in pharyngeal and laryngeal pathologies in patients with proven gastric colonization. Head Neck 2013, 36, 1562–1566. [Google Scholar] [CrossRef]
- Nártová, E.; Kraus, J.; Pavlik, E.; Lukeš, P.; Katra, R.; Plzák, J.; Kolářová, L.; Šterzl, I.; Betka, J.; Astl, J. Presence of different genotypes of Helicobacter pylori in patients with chronic tonsillitis and sleep apnoea syndrome. Eur. Arch. Oto-Rhino-Laryngol. 2014, 271, 607–613. [Google Scholar] [CrossRef]
- Rotnáglová, E.; Tachezy, R.; Salakova, M.; Procházka, B.; Košl’abová, E.; Vesela, E.; Ludvíková, V.; Hamšíková, E.; Klozar, J. HPV involvement in tonsillar cancer: Prognostic significance and clinically relevant markers. Int. J. Cancer 2011, 129, 101–110. [Google Scholar] [CrossRef]
- Berman, T.A.; Schiller, J.T. Human papillomavirus in cervical cancer and oropharyngeal cancer: One cause, two diseases. Cancer 2017, 123, 2219–2229. [Google Scholar] [CrossRef] [PubMed]
- Simonidesova, S.; Hamsikova, E.; Ludvikova, V.; Klozar, J.; Vencalek, O.; Tachezy, R. Prognostic value of posttreatment HPV-specific antibodies in patients with oropharyngeal tumors. J. Surg. Oncol. 2019, 120, 117–124. [Google Scholar] [CrossRef]
- Timbang, M.R.; Sim, M.W.; Bewley, A.F.; Farwell, D.G.; Mantravadi, A.; Moore, M.G. HPV-related oropharyngeal cancer: A review on burden of the disease and opportunities for prevention and early detection. Hum. Vaccines Immunother. 2019, 15, 1920–1928. [Google Scholar] [CrossRef] [PubMed]
- D’Souza, G.; McNeel, T.; Fakhry, C. Understanding personal risk of oropharyngeal cancer: Risk-groups for oncogenic oral HPV infection and oropharyngeal cancer. Ann. Oncol. 2017, 28, 3065–3069. [Google Scholar] [CrossRef] [PubMed]
- Wittekindt, C.; Klussmann, J.P. Tumor Staging and HPV-Related Oropharyngeal Cancer. Adv. Struct. Saf. Stud. 2016, 206, 123–133. [Google Scholar] [CrossRef]
- Lukeš, P.; Pavlik, E.; Potužníková, B.; Plzák, J.; Nártová, E.; Doseděl, J.; Katra, R.; Šterzl, I.; Betka, J.; Astl, J. Comparison of Helicobacter Pylori Genotypes Obtained from the Oropharynx and Stomach of the Same Individuals–A Pilot Study. Prague Med. Rep. 2012, 113, 231–239. [Google Scholar] [CrossRef]
- Lukeš, P.; Pavlik, E.; Potuznikova, B.; Nartova, E.; Foltynova, E.; Plzák, J.; Katra, R.; Sterzl, I.; Bartunkova, J.; Betka, J.; et al. Detection of Helicobacter pylori in oropharyngeal lymphatic tissue with real-time PCR and assessment of its carcinogenic potential. Eur. Arch. Oto-Rhino-Laryngol. 2014, 271, 399–405. [Google Scholar] [CrossRef]
- Katra, R.; Kabelka, Z.; Jurovcik, M.; Hradsky, O.; Kraus, J.; Pavlik, E.; Nartova, E.; Lukeš, P.; Astl, J. Pilot study: Association between Helicobacter pylori in adenoid hyperplasia and reflux episodes detected by multiple intraluminal impedance in children. Int. J. Pediatric Otorhinolaryngol. 2014, 78, 1243–1249. [Google Scholar] [CrossRef] [PubMed]
- Chen, J.; Domingue, J.C.; Sears, C.L. Microbiota dysbiosis in select human cancers: Evidence of association and causality. Semin. Immunol. 2017, 32, 25–34. [Google Scholar] [CrossRef]
- Lopez, E.M.; Tanner, A.M.; Du, E.; Patel, S.N.; Weiss, J.; Weissler, M.C.; Hackman, T.; Gupta, G.P.; Zevallos, J.; Elmore, S.; et al. Decline in circulating viral and human tumor markers after resection of head and neck carcinoma. Head Neck 2021, 43, 27–34. [Google Scholar] [CrossRef]
- De Martel, C.; Ferlay, J.; Franceschi, S.; Vignat, J.; Bray, F.; Forman, D.; Plummer, M. Global burden of cancers attributable to infections in 2008: A review and synthetic analysis. Lancet Oncol. 2012, 13, 607–615. [Google Scholar] [CrossRef]
- Ang, K.K.; Harris, J.; Wheeler, R.; Weber, R.; Rosenthal, D.I.; Nguyen-Tân, P.F.; Westra, W.H.; Chung, C.H.; Jordan, R.C.; Lu, C.; et al. Human Papillomavirus and Survival of Patients with Oropharyngeal Cancer. N. Engl. J. Med. 2010, 363, 24–35. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Payão, L.T.R.S.L.M. Helicobacter pyloriand its reservoirs: A correlation with the gastric infection. World J. Gastrointest. Pharmacol. Ther. 2016, 7, 126–132. [Google Scholar] [CrossRef]
- Hwang, M.S.; Forman, S.N.; Kanter, J.A.; Friedman, M. Tonsillar Helicobacter pylori Colonization in Chronic Tonsillitis. JAMA Otolaryngol. Neck Surg. 2015, 141, 245. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Siupsinskiene, N.; Katutiene, I.; Jonikiene, V.; Janciauskas, D.; Vaitkus, S. Helicobacter pylori in the tonsillar tissue: A possible association with chronic tonsillitis and laryngopharyngeal reflux. J. Laryngol. Otol. 2017, 131, 549–556. [Google Scholar] [CrossRef]
- Najafipour, R.; Ahmadpour, F.; Farivar, T.N.; Pahlevan, A.; Johari, P.; Safdarian, F.; Mehr, M.A. Assessment of Helicobacter pylori prevalence by scorpion real-time PCR in chronic tonsillitis patients. J. Glob. Infect. Dis. 2012, 4, 38–42. [Google Scholar] [CrossRef]
- Song, Q.; Lange, T.; Spahr, A.; Adler, G.; Bode, G. Characteristic distribution pattern of Helicobacter pylori in dental plaque and saliva detected with nested PCR. J. Med. Microbiol. 2000, 49, 349–353. [Google Scholar] [CrossRef] [PubMed]
- Tang, K.D.; Vasani, S.; Menezes, L.; Taheri, T.; Walsh, L.J.; Hughes, B.G.; Frazer, I.H.; Kenny, L.; Scheper, G.C.; Punyadeera, C. Oral HPV16 DNA as a screening tool to detect early oropharyngeal squamous cell carcinoma. Cancer Sci. 2020, 111, 3854–3861. [Google Scholar] [CrossRef] [PubMed]
- Hatakeyama, M. Structure and function of Helicobacter pylori CagA, the first-identified bacterial protein involved in human cancer. Proc. Jpn. Acad. Ser. B 2017, 93, 196–219. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Basso, D.; Navaglia, F.; Brigato, L.; Piva, M.G.; Toma, A.; Greco, E.; Di Mario, F.; Galeotti, F.; Roveroni, G.; Corsini, A.; et al. Analysis of Helicobacter pylori vacA andcagA genotypes and serum antibody profile in benign and malignant gastroduodenal diseases. Gut 1998, 43, 182–186. [Google Scholar] [CrossRef] [Green Version]
- Miehlke, S.; Kirsch, C.; Agha-Amiri, K.; Günther, T.; Lehn, N.; Malfertheiner, P.; Stolte, M.; Ehninger, G.; Bayerdörffer, E. The Helicobacter pylori vacA s1, m1 genotype and cagA is associated with gastric carcinoma in Germany. Int. J. Cancer 2000, 87, 322–327. [Google Scholar] [CrossRef]
- Fahimi, F.; Tohidkia, M.R.; Fouladi, M.; Aghabeygi, R.; Samadi, N.; Omidi, Y. Pleiotropic cytotoxicity of VacA toxin in host cells and its impact on immunotherapy. BioImpacts 2017, 7, 59–71. [Google Scholar] [CrossRef] [PubMed]
- Chauhan, N.; Tay, A.C.Y.; Marshall, B.J.; Jain, U. Helicobacter pyloriVacA, a distinct toxin exerts diverse functionalities in numerous cells: An overview. Helicobacter 2019, 24, e12544. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Caston, R.R.; Sierra, J.C.; Foegeding, N.J.; Truelock, M.D.; Campbell, A.M.; Frick-Cheng, A.E.; Bimczok, D.; Wilson, K.T.; McClain, M.S.; Cover, T.L. Functional Properties of Helicobacter pylori VacA Toxin m1 and m2 Variants. Infect. Immun. 2020, 88, e00032-20. [Google Scholar] [CrossRef]
Total | HP+ | % | p-Value | HPV+ | % | p-Value | |
---|---|---|---|---|---|---|---|
Malignancy Group A | 103 | 86 | 83.5 | <0.001 | 62 | 60.2 | 0.006 |
Inflammation Group B | 85 | 65 | 76.5 | 38 | 44.7 | ||
Control Group C | 50 | 6 | 12.0 | 17 | 34.0 |
Total | HP+ | CagA+ | % of Total | CagA− | % of Total | |
---|---|---|---|---|---|---|
Malignancy Group A | 103 | 86 | 24 | 23.3 | 62 | 60.2 |
Inflammation Group B | 85 | 65 | 13 | 15.3 | 52 | 61.2 |
Control Group C | 50 | 6 | 2 | 4.0 | 4 | 8.0 |
Total HP+ | CagA+ | % | CagA− | % | p-Value | |
---|---|---|---|---|---|---|
Malignancy | 86 | 24 | 27.9 | 62 | 72.1 | 0.482 |
Group A | ||||||
Inflammation | 65 | 13 | 20.0 | 42 | 80.0 | |
Group B | ||||||
Control | 6 | 2 | 33.3 | 4 | 66.7 | |
Group C |
Malignancy (n =103) | Total | % |
HP−/HPV− | 10 | 9.7 |
HP−/HPV+ | 7 | 6.8 |
HP-CagA+/HPV- | 16 | 15.5 |
HP-CagA−/HPV− | 17 | 16.5 |
HP-CagA+/HPV+ | 8 | 7.8 |
HP-CagA−/HPV+ | 45 | 43.7 |
Chronic Inflammation (n = 85) | Total | % |
HP−/HPV− | 13 | 15.3 |
HP−/HPV+ | 7 | 8.2 |
HP-CagA+/HPV− | 8 | 9.4 |
HP-CagA−/HPV− | 23 | 27.1 |
HP-CagA+/HPV+ | 6 | 7.1 |
HP-CagA−/HPV+ | 28 | 32.9 |
Control Group (n = 50) | Total | % |
HP−/HPV− | 30 | 60.0 |
HP−/HPV+ | 14 | 28.0 |
HP-CagA+/HPV− | 1 | 2.0 |
HP-CagA−/HPV− | 2 | 4.0 |
HP-CagA+/HPV+ | 1 | 2.0 |
HP-CagA−/HPV+ | 2 | 4.0 |
CagA Protein Gene Detection | Genotype of VacA Cytotoxin | DETECTED (N) | % |
---|---|---|---|
POSITIVE | VacA S1a M1 | 4 | 4.7 |
POSITIVE | VacA S1a M2 | 5 | 5.8 |
POSITIVE | VacA S1b M1 | 10 | 11.6 |
POSITIVE | VacA S1b M2 | 4 | 4.7 |
POSITIVE | VacA S2 M1 | 1 | 1.2 |
POSITIVE | VacA S2 M2 | 0 | 0.0 |
NEGATIVE | VacA S1a M1 | 17 | 19.8 |
NEGATIVE | VacA S1a M2 | 11 | 12.8 |
NEGATIVE | VacA S1b M1 | 19 | 22.1 |
NEGATIVE | VacA S1b M2 | 10 | 11.6 |
NEGATIVE | VacA S2 M1 | 5 | 5.8 |
NEGATIVE | Vac A S2 M2 | 0 | 0.0 |
CagA Protein Gene Detection | Genotype of VacA Cytotoxin | DETECTED (N) | % |
---|---|---|---|
POSITIVE | VacA S1a M1 | 8 | 12.3 |
POSITIVE | VacA S1a M2 | 1 | 1.5 |
POSITIVE | VacA S1b M1 | 7 | 10.8 |
POSITIVE | VacA S1b M2 | 1 | 1.5 |
POSITIVE | VacA S2 M1 | 0 | 0.0 |
POSITIVE | VacA S2 M2 | 0 | 0.0 |
NEGATIVE | VacA S1a M1 | 9 | 13.9 |
NEGATIVE | VacA S1a M2 | 10 | 15.4 |
NEGATIVE | VacA S1b M1 | 11 | 16.9 |
NEGATIVE | VacA S1b M2 | 12 | 18.5 |
NEGATIVE | VacA S2 M1 | 6 | 9.2 |
NEGATIVE | VacA S2 M2 | 0 | 0.0 |
CagA Protein Gene Detection | Genotype of VacA Cytotoxin | DETECTED (N) |
---|---|---|
POSITIVE | VacA S1a M1 | 1 |
POSITIVE | VacA S1a M2 | |
POSITIVE | VacA S1b M1 | |
POSITIVE | VacA S1b M2 | |
POSITIVE | VacA S2 M1 | 1 |
POSITIVE | VacA S2 M2 | |
NEGATIVE | VacA S1a M1 | 1 |
NEGATIVE | VacA S1a M2 | 1 |
NEGATIVE | VacA S1b M1 | 1 |
NEGATIVE | VacA S1b M2 | 1 |
NEGATIVE | VacA S2 M1 | |
NEGATIVE | VacA S2 M2 |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Astl, J.; Holy, R.; Maute, E.; Rotnágl, J.; Kalfeřt, D.; Drnková, B.; Younus, T.; Pavlík, E. Genome of Helicobacter pylori and Serotype of HPV Detected in Oropharyngeal and Laryngeal Cancer and Chronic Inflammation Patients. Int. J. Environ. Res. Public Health 2021, 18, 9545. https://doi.org/10.3390/ijerph18189545
Astl J, Holy R, Maute E, Rotnágl J, Kalfeřt D, Drnková B, Younus T, Pavlík E. Genome of Helicobacter pylori and Serotype of HPV Detected in Oropharyngeal and Laryngeal Cancer and Chronic Inflammation Patients. International Journal of Environmental Research and Public Health. 2021; 18(18):9545. https://doi.org/10.3390/ijerph18189545
Chicago/Turabian StyleAstl, Jaromír, Richard Holy, Eva Maute, Jan Rotnágl, David Kalfeřt, Barbora Drnková, Temoore Younus, and Emil Pavlík. 2021. "Genome of Helicobacter pylori and Serotype of HPV Detected in Oropharyngeal and Laryngeal Cancer and Chronic Inflammation Patients" International Journal of Environmental Research and Public Health 18, no. 18: 9545. https://doi.org/10.3390/ijerph18189545
APA StyleAstl, J., Holy, R., Maute, E., Rotnágl, J., Kalfeřt, D., Drnková, B., Younus, T., & Pavlík, E. (2021). Genome of Helicobacter pylori and Serotype of HPV Detected in Oropharyngeal and Laryngeal Cancer and Chronic Inflammation Patients. International Journal of Environmental Research and Public Health, 18(18), 9545. https://doi.org/10.3390/ijerph18189545