No abstract

Integration of the human papillomavirus (HPV) genome is thought to be one of the causes of cancer progression. However, there is controversy concerning the physical status of HPV 16 in premalignant cervical lesions, and there have been no reports on the concordance between detection of the integrated form of HPV16 by real-time PCR and by in situ hybridization. We investigated specimens of cervical intraepithelial neoplasia (CIN) and invasive carcinomas for the physical status of HPV 16 by real-time PCR and in situ hybridization. The presence of the integrated form was detected by both real-time PCR and in situ hybridization in zero of four cases of CIN1, three of six cases of CIN2, nine of 27 cases of CIN3, and two of six cases of invasive carcinomas. Integrated HPV 16 was present in some premalignant lesions but was not always present in carcinomas. The concordance rate between the two methods for the detection of the presence of the integrated form was 37 of 43 (86%) cases. Real-time PCR and in situ hybridization were found to be complementary and convenient techniques for determining the physical status of the HPV genome. We conclude that a combination of both methods is a more reliable means of assessing the physical status of the HPV genome in cervical neoplasia.

As human papillomavirus (HPV) infection is the main risk factor for squamous cell carcinoma of the cervix and overexpression of p16INK4a occurs when retinoblastoma protein is inactivated by high-risk HPV, the authors studied the association of HPV infection and expression of p16INK4a in cervical adenocarcinomas. Specimens of cervical glandular neoplasias were immunostained with a p16INK4a-specific monoclonal antibody (clone E6H4). Approximately 80% of glandular neoplasms showed overexpression of p16INK4a. Exfoliated cells from 14 adenocarcinomas were further examined by p16INK4a-specific immunocytochemistry, and 12 cases showed overexpression of p16INK4a, suggesting that immunostaining for p16INK4a may be a useful diagnostic tool for cervical adenocarcinomas. The authors further examined HPV DNA in cervical adenocarcinomas with the polymerase chain reaction method. Overexpression of p16INK4a was positive in 94% of cases in which HPV16 or 18DNA was positive, a finding suggesting that HPV16 or 18 may play an important role in cervical adenocarcinomas. Overexpression of p16INK4a may be an indicator of pathogenic activity of high-risk HPVs.

A fluid-based Papanicolaou test has been established to improve sample collection and preparation. This study was the first largescale investigation in Japan to examine the feasibility of using fluid-based Papanicolaou specimens to detect human papillomavirus (HPV) using Hybrid Capture II and polymerase chain reaction (PCR). Three thousand patients who visited Keio University Hospital between October 2000 and February 2001 were enrolled in the study. The results of the fluid-based Papanicolaou tests corresponded well with those of conventional Papanicolaou smears (96.8% concordance). The sensitivities of cervical neoplasia detection using the fluid-based Papanicolaou test (73.9%) and Hybrid Capture II (76.3%, P ¼ 0.55) were not significantly different. Among the cervical intraepithelial neoplasia 3 and squamous cell carcinoma specimens, HPV 16 and HPV 52 were predominantly detected using the PCR method. Although some DNA samples extracted from the fluid-based specimens were degradaded, PCR and direct sequencing could be performed without difficulty even after 1 year of specimen storage. We conclude that fluid-based Papanicolaou specimens can be applied to investigate HPV infection. The fluid-based Papanicolaou specimen collection method is widely used for primary cervical cancer screening (Awen et al, 1994;Roberts et al, 1997;Bolick and Hellman, 1998), largely because of the improvement in specimen quality, the advantage of increased sensitivity, and a reduction in the false negative rate for squamous intraepithelial lesions (Linder and Zahniser, 1997;Park et al, 2001), compared to the conventional Papanicolaou smear method. Fluid-based specimens can be stored at ambient temperature for a longer period; therefore, this collection system offers the advantage that once the specimen has been collected, there is no need to collect additional specimens from the patient to perform a second Papanicolaou test or conduct further investigations. Studies evaluating the clinical utility of Hybrid Capture II (HC II) human papillomavirus (HPV)-DNA testing have also been performed (Clavel et al, 1998;Lin CT et al, 2000a;Yamazaki et al, 2001;Castle et al, 2002). Genital HPV has been reported to be related to cervical cancer carcinogenesis and some types of HPV; HPV 16, for example, is associated with a high risk of cervical neoplasia (Josefsson et al, 2000;Woodman et al, 2001). HC II is a HPV detection test designed to detect 18 types of HPV using microtitre plates and is an appropriate method for HPV screening. We performed the HC II test using fluid-based specimens from patients in whom biopsy studies were also performed to determine whether this method is appropriate for detecting cervical neoplasia in Japan. We also performed HPV typing using fluid-based specimens. We previously reported an HPV-DNA transcript detection method using cytologic specimens and reverse transcriptase-nested polymerase chain reactions (PCR) (Fujii et al, 1995), and a method for detecting multiplex HPV infection using PCR single-stranded DNA-conform...