Aberrant DNA methylation of DLX4 and SIM1 is a predictive marker for disease progression of uterine cervical low-grade squamous intraepithelial lesion.

BACKGROUND: A few uterine cervical low-grade squamous intraepithelial lesions (LSILs) are known to progress with high-risk human papillomavirus (hrHPV). METHODS: One hundred and thirteen patients were classified into four groups according to their cervical cytology, hrHPV infection, and follow up. Cytology samples were examined for aberrant DNA methylation of DLX4 and SIM1 genes and protein expressions. CaSki cells were treated with 5-Aza-2'-deoxycytidine (5-aza-dC). RESULTS: Group 1 was negative for intraepithelial lesions or malignancies. LSIL in group 2 showed a continuance of LSIL for longer than 365 days and LSIL in group 3 showed an upgrading to high-grade (H) SIL or higher (HSIL+) within 365 days of LSIL diagnosis. Group 4 was squamous cell carcinoma. All but group 1 were infected with hrHPV. Significant differences existed in the frequency of DNA methylation between groups 2 and 3 (p = 0.044), between groups 3 and 4 (p = 0.020) for DLX4, and between groups 1 and 3 (p = 0.0003), and groups 2 and 3 (p = 0.005) for the SIM1 gene. DLX4 protein expression was significantly reduced in the DLX4 methylation positive tissues (p = 0.008). The 5-aza-dC treatment restored DLX4 mRNA expressions of CaSki cells (p < 0.005). The LSIL cases with DNA methylation of the SIM1 gene, or both genes, progressed faster to HSIL+ than did the others (p = 0.033 and p = 0.045, respectively). CONCLUSION: Aberrant DNA methylation of the DLX4 and SIM1 genes should be a novel progression marker for uterine cervical LSIL with hrHPV infection.

[Auto-analysis of immunohistochemical findings for breast cancer using specified software and virtual microscopy].

Currently, the therapeutic strategy for a breast cancer patient is designed according to their histopathological, immunohistochemical and molecular findings. These findings are obtained through the collected efforts of many individual pathologists or medical technologists (MTs) and are, thus, limited by intra-observer error and potentially subjective decision making. Twenty five breast cancer specimens collected between November 2009 and February 2010 were examined for immunohistochemical expressions of estrogen receptor (ER), progesterone receptor (PgR), HER2, Ki-67, Topoisomerase II alpha (Topo IIalpha). Fifty one cancer specimens collected November 2009 and June 2010 were examined for human epidermal growth factor 2 (HER2). Immunohistochemical staining was performed using auto-stainers (Ventana) and the results were stored digitally after examination by virtual microscopy (Hamamatsu Photonics). Data analysis was performed with the Genie/Aperio software package on a desk-top computer. For all the antibodies used expect for HER2, concordant results were obtained in 100% of 24 ER positive cases. Ki-67 index (r=0.96) and Topo IIalpha index (r=0.95) also showed a significant correlation (p<0.001). For HER2, all four specimens with Hercep-score 2 by ocular observation but auto-analysis score 1 revealed no HER2 gene amplification. Well-organized auto-analysis is more likely to result in an objective observation and to provide a means by which to standardize the methods for immunohistochemical detection of breast cancer.

Histological correlation of glandular abnormalities in cervical liquid-based cytology.

Conventional Papanicolaou smear method is still commonly used for cervical cancer screening in Japan, despite the liquid-based cytology (LBC) that has become a global tendency in the world recently. One of the obstacles in the way of popularization of this method seems to be the confusion as to diagnosis upon cervical glandular lesions. We performed comparison study between LBC and conventional Papanicolaou smear about cytological diagnosis using split-sample method in 4522 patients. In 13 cases analyses, which were reported with either AGC or adenocarcinoma by either method, LBC tends to be milder than that by conventional smear, however, the credibility of LBC is considered to be near to that of conventional smear with regard to screening for glandular abnormalities. These results indicate that cervical cancer screening should shift to LBC under the enough experience and appropriate dealing with the cytological diagnosis.

Human papillomavirus types 52 and 58 are prevalent in uterine cervical squamous lesions from Japanese women.

Objective. To estimate the prevalence and genotypes of high-risk human papillomavirus (HPV) focusing HPV 16, 18, 52, and 58 in Japan. Methods. Liquid-base cytology specimens were collected from Japanese women (n = 11022), aged 14-98. After classifying cytodiagnosis, specimens were analyzed for HPV DNA by the multiplex polymerase chain reaction method, where 1195 specimens were positive for cervical smear, except adenomatous lesions. Result. HPV genotypes were detected in 9.5% of NILM and 72.2% of ASC-US or more cervical lesions. In positive cervical smears, HPV genotypes were HPV 52 at 26.6%, HPV 16 at 25.2%, HPV 58 at 21.8%, and HPV 18 at 7.1%. Most patients infected with HPV 16 were between 20-29 years old, decreasing with age thereafter. As for HPV 52 and 58, although the detection rate was high in 30- to 39-year-olds, it also was significant in the 50s and 60s age groups. Conclusion. In Japan, as a cause of abnormal cervical cytology, HPV52 and 58 are detected frequently in addition to HPV 16. In older age groups, HPV 52 and 58 detection rates were higher than that observed for HPV 16. After widespread current HPV vaccination, we still must be aware of HPV 52 and 58 infections.