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Human papillomavirus (HPV) is an independent risk factor for development of oral squamous cell carcinoma (OSCC). This study aimed to investigate the role of HPV infection and the trend in percentage of HPV-associated OSCC over a 5-year period in northeastern Thailand. In this case-control study, 91 exfoliated oral cell samples and 80 lesion cell samples from OSCC cases and exfoliated oral cells from 100 age/gender-matched controls were collected. HPV infection was investigated by PCR using GP5+/GP6+ primers followed by HPV genotyping using reverse line blot hybridization. Quantitative RT-PCR was used to evaluate HPV oncogene transcription. Temporal trends of HPV infection were evaluated in archived formalin-fixed paraffin-embedded (FFPE) OSCC tissues using in situ hybridization. HPV DNA was found in 17.5% (14/80) of lesion samples from OSCC cases and 29.7% (27/91) of exfoliated oral cell samples from the same cases. These values were significantly higher than in exfoliated oral cell samples from controls (13%, 13/100). HPV-16 was the genotype most frequently found in OSCC cases (92.8%, 13/14 infected cases). Interestingly, HPV oncogene mRNA expression was detected and correlated with OSCC cases (P < 0.005). Of 146 archived FFPE OSCC samples, 82 (56.2%) were positive for high-risk HPV DNA and 64 (43.8%) cases were positive for HPV E6/E7 mRNA expression. There was a trend of increasing percentage of HPV-associated OSCC from 2005 to 2010. This was especially so for females with well-differentiated tumors in specific tongue sub-sites. We suggest that HPV infection plays an important role in oral carcinogenesis in northeastern Thailand., (© 2016 Wiley Periodicals, Inc.)

To understand the potential role in cervical cancer development of the three most common high-risk human papillomavirus (HR-HPVs) in Thai women, HPV genotypes and viral genome statuses in different cervical lesions were investigated. Cervical tissues consisting of no cervical intraepithelial neoplasia (84 cases), grade I cervical intraepithelial neoplasia (176 cases), grade II-III cervical intraepithelial neoplasia (91 cases), and squamous cell carcinoma (66 cases) were subjected for HPV genotyping by polymerase chain reaction (PCR) and reverse line blot hybridization assay and for HPV genome status determination by amplification of papillomavirus oncogene transcripts (APOT) assay. HPV prevalence was 28.6% in no cervical intraepithelial neoplasia, 40.3% in grade I cervical intraepithelial neoplasia, 70.3% in grade II-III cervical intraepithelial neoplasia and 86.4% in squamous cell carcinoma cases. The three most common HR-HPV types were HPV 16, 58, and 18 which were distributed in all cervical lesions. HPV physical statuses could be investigated in 4 no cervical intraepithelial neoplasias, 2 grade I cervical intraepithelial neoplasias, 28 grade II-III cervical intraepithelial neoplasias and 31 squamous cell carcinomas. The integrated-derived transcripts were found 3.6% in grade II-III cervical intraepithelial neoplasia and 48.4% in squamous cell carcinoma, whereas no viral genome integration was found in the group of no cervical intraepithelial neoplasia or grade I cervical intraepithelial neoplasia samples. The frequencies of HR-HPV integration in squamous cell carcinoma were found 40%, 100%, 20% of HPV 16, 18, and 58. This study indicates the oncogenic potential ability of the three most common HR-HPVs associated with cervical cancer progression., (© 2014 Wiley Periodicals, Inc.)

HPV 58 is detected commonly in cervical cancer in East Asian countries. To evaluate the HPV 58 physical state, the amplification of papillomavirus oncogene transcripts (APOT) and hybridisation assays were established. Episome- and integrate-derived transcripts were confirmed by direct sequencing. Twenty-nine HPV 58 positive samples from various cervical lesions were used. The results showed that the episome-derived transcripts were recognised as two major specific amplified products (1040 and 714 bp). Two splice donor sites were mapped to the 5' splice site of the E1 gene on SD898 and SD899 and spliced to the 3' acceptor site of the E4 gene on SA3353, SA3356 and SA3365. The episome-derived transcripts were found 100% in normal cervical epithelia and low-grade lesions (9/9 cases) while the integrate-derived transcripts were detected in 13.3% of high-grade lesions (2/15 cases) and in 20% of carcinomas (1/5 cases). HPV 58 integration sites were found on chromosomes 4q21, 12q24 and 18q12. Using the established APOT assay, the results revealed not only novel information on the HPV 58 transcription patterns of episomal transcripts, but also integration site. The APOT assay is a reliable and useful tool for the detection of the HPV 58 physical state and its oncogene expression., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Aim: This study investigated the utility of visual inspection with acetic acid (VIA) as a method for cervical cancer screening in Thailand and examined the relationship of VIA to high-risk human papillomavirus (HR-HPV) status., Methods: Cervical cells were collected from 160 patients receiving a Pap smear. VIA was performed on the cervix of the patients by application of 5% acetic acid. HPV screening of DNA extracted from cytology samples was performed by PCR using the GP5+/6+ primer system followed by reverse line blot hybridization genotyping., Results: The majority (96.9%) of the patients were diagnosed with normal or inflammatory cytologic changes. 32.8% of normal cytology and 42.0% of inflammation cases showed positive acetowhite staining. 3.1%, 38.1% and 42.5% of subjects were positive for an abnormal Pap test, VIA test, and HPV DNA, respectively. VIA demonstrated 50% sensitivity and 66.7% specificity for abnormal histology with PPV and NPV values of 50% and 66.7%, respectively, whereas HPV DNA test showed 100% sensitivity. HPV16 was the most common (54.4%) and HR-HPV was detected in 36.3% of all cases. 48.5% of HR-HPV positive and 36.8% of HR-HPV negative cervices stained with acetowhite following the VIA test., Conclusion: The VIA test is a simple method for cervical cancer screening; however, a significant proportion of patients with normal or inflammatory cytology were positive by this test. Further, HR-HPV in women without acetowhite staining was demonstrated. Therefore, some form of HR-HPV detection test may be required for combination with cervical cell screening even in low-resource nations., (© 2010 The Authors. Journal of Obstetrics and Gynaecology Research © 2010 Japan Society of Obstetrics and Gynecology.)

Objective: To evaluate the value of the combination of p16 and HPV detection in the screening for cervical cancer., Methods: 186 patients with previous abnormal cervical lesion were studied. After colposcopic examination, two conventional Pap slides were prepared: the first was Papanicolaou-stained and examined by cytologist; the second was immunocytochemically stained for p16. Cervical cells were collected by brush using for HPV detection by Hybrid Capture II. Biopsy of any colposcopically abnormal lesions was performed., Results: The 186 cervical samples were classified cytologically as normal (148), ASCUS (13), low-grade (11), high-grade (12) dysplasia and squamous cell carcinoma (2). P16 and HPV were found in all high-grade dysplasia and SCC, and in 64% and 27% of low-grade dysplasia, 62% and 0% of ASCUS and 7.4% and 3.4% of normal, respectively. 18 of p16-positive cases (11%) were HPV-negative, 14 of them in the ASCUS and normal group. Compared to histological results, all of the p16-positive cases of squamous metaplasia, CIN II/III and SCC were HR-HPV-positive. Therefore, the cases that were positive for both with normal cytology (5 cases) or low-grade dysplasia (3 cases) may comprise a high-risk group for neoplastic change., Conclusion: The combination of p16 and HPV detection may be useful in cervical cancer screening to identify high-risk patients requiring early and proper management.

The p16INK4a protein is immunocytochemically detected in liquid-based (LB) specimens as a diagnostic marker of cervical dysplasia and neoplasia. Its up-regulation is promoted by high-risk human papillomavirus (HR-HPV) infection. We aimed to detect p16INK4a on conventional Papanicolaou (Pap) test (CPT) slides and to determine the relationship between its overexpression and HR-HPV infection. CPT and LB Pap test (LBPT) slides (165 samples of each) were examined by immunocytochemical staining for p16INK4a. After polymerase chain reaction (PCR), HPV-DNA was genotyped by dot blot hybridization. The CPT slides displayed more numerous dispersed squamous cells and LBPT slides had a clearer background. Positive p16INK4a on CPT occurred in 0% (0/30), 52.5% (21/40), 54.3% (19/35), 100% (30/30), and 100% (30/30) in normal, atypical squamous cells of undetermined significance (ASCUS), low-grade squamous intraepithelial lesions (LSILs), high-grade SILs (HSILs), and squamous cell carcinomas (SCCs) cases, respectively. LBPT slides showed comparable results but were less sensitive. HPV-DNA was detected in 86.7, 70, 45, 57.14, and 10% in SCCs, HSILs, ASCUS, LSILs, and normal cervical cells, respectively. Because HR-HPV was identified in all HPV+ samples of high-grade dysplasia (HSILs and SCCs) and all positive p16INK4a samples infected with HR-HPV, the association of p16INK4a overexpression with HR-HPV infection was confirmed. This study suggests that immunocytochemical staining of p16INK4a on CPT slides is convenient and cost-effective for cervical cancer screening by the detection of dysplastic cells infected with HR-HPV., (2004 Wiley-Liss, Inc.)

Objective: To correlate the detection of human papillomavirus (HPV) with the Pap smear classification of cervical-scraped cells from asymptomatic women living in northeast Thailand., Methods: A total of 260 asymptomatic women attending the Obstetrics and Gynecology's Outpatient Clinic, Srinagarind Hospital, Thailand, were interviewed for risk factors and cervical scrapes were taken. The cells were examined by Pap smear for cytological changes and by PCR for HPV DNAs--nononcogenic (HPVs 6 and 11) and oncogenic (HPVs 16, 18 and 33) types. Cervical biopsies were taken from women with abnormal Pap smears for histological examination., Results: Of the 260 cervical smear samples, the cervical cells were classified as normal and abnormal in 174 and 86, respectively. Twenty-three percent of all samples were positive for overall HPV DNA. HPV DNAs (mostly HPVs 6 and 11) were detected in 21% of normal cells, and the higher detection rate (27%) for HPV DNA in abnormal cells gradually increased in severity from 16% in Class 3 to 35 and 60% in Classes 4 and 5, respectively. Histologically 46, 90 and 100% of HPV detection was associated with CIS, SCC and adenocarcinoma, respectively. Almost all of the HPV DNAs detected were types 16, 18 or 33. There was no significant association between HPV infection and reproduction history, sexual behavior and demographic variables., Conclusion: We speculated that an abnormal Pap smear and the detection of an oncogenic type HPV may indicate the presence of neoplastic cells in asymptomatic woman who might be at risk for the development of cervical cancer.