Abstract
Incidence of human papillomavirus (HPV)-associated oral cancers is on the rise. However, epidemiological data of this subset of cancers are limited. Dental hospital poses a unique advantage in detection of HPV-positive oral malignancies. We assessed the utility of formalin-fixed paraffin-embedded (FFPE) tissues, which are readily available, for evaluation of high-risk HPV infection in oral cancer. For protocol standardization, we used 20 prospectively collected paired FFPE and fresh tissues of histopathologically confirmed oral cancer cases reported in Oral Medicine department of a dental hospital for comparative study. Only short PCRs (~ 200bp) of DNA isolated using a modified xylene-free method displayed a concordant HPV result. For HPV analysis, we used additional 30 retrospectively collected FFPE tissues. DNA isolated from these specimens showed an overall 23.4% (11/47) HPV positivity with detection of HPV18. Comparison of HPV positivity from dental hospital FFPE specimens with overall HPV positivity of freshly collected oral cancer specimens (n = 55) from three cancer care hospitals of the same region showed notable difference (12.7%; 7/55). Further, cancer hospital specimens showed HPV16 positivity and displayed a characteristic difference in reported sub-sites and patient spectrum. Overall, using a xylene-free FFPE DNA isolation method clubbed with short amplicon PCR, we showed detection of HPV-positive oral cancer in dental hospitals.
Highlights
India harbors the world’s largest number of oral cancer cases [1, 2]
Prior to detection of Human Papillomavirus (HPV) infection in oral cancer by PCR using consensus degenerate sequence-specific primers PGMY09/PGMY11 and GP5+/GP6+ and its typing the adequacy of DNA was assessed by subjecting the genomic DNA (100 ng/reaction) isolated from above-mentioned methods for β-globin or p53 Exon 5 PCR which were used as internal controls targeted to host genomic DNA
All samples were further processed for HPV genotyping by HPV type-specific PCR for HPV16 and HPV18 with short amplicon sizes as described previously [36, 37]
Summary
India harbors the world’s largest number of oral cancer cases [1, 2]. Oral cancers are among the largest group of preventable cancers. Regular screening and prevention of high-risk habits like smoking, tobacco chewing, and consumption of alcohol and areca nut could effectively control them [3] These cancers are biologically and psychologically the worst when diagnosed at an advanced stage. Infection of highrisk HPVs is becoming a significant contributor, a decisive risk factor, and likely an etiological agent for these subsets of oral cancers [5, 6], S.19 [7]. These cancers occur early in age and have no prior history of tobacco and/or high alcohol consumption [4]. More than 90% of HPV-positive oral cancer are infected predominantly with the HPV16 genotype [17], whereas the presence of HPV18 has been reported only sporadically in a few specific studies [8, 18,19,20,21,22]
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