DNA quantification is technically feasible and of value in cervical smear samples: possible applications for determination of progression in low grade dyskaryosis

Abstract

This study evaluates the feasibility of DNA analysis of cervical intraepithelial neoplasia III (CIN III) lesions on cervical smear and formalin-fixed paraffin-embedded tissue (FFPET) blocks with a view to extending this type of analysis to milder grades of dyskaryosis. DNA ploidy was determined by image analysis using a CAS 200 Image Analyser. Seventeen patients with a diagnosis of CIN III were studied. Results show that all smear and tissue samples were non-diploid with nine aneuploid and eight tetraploid lesions. In 6/7 patients whose smears and corresponding biopsies were examined there was complete agreement as to the DNA profile. We conclude that DNA quantification is technically feasible in archival, routinely prepared cervical smears. This technique should now be applied to CINI and CINII cervical smears to determine if it is of value in identifying those lesions that will progress to CIN III. This study is particularly timely with the possibility in the near future of estimation of ploidy by image analysis using instruments such as the Highly Optimized Microscope Environment (HOME) system.

Diagnostic and prognostic significance of image cytometric DNA ploidy measurement in cytological samples of cervical squamous intraepithelial lesions

“…All 17 cases of CIN3 reported by Walsh et al. 21 had non‐diploid DNA content; nine were aneuploid and eight were tetraploid. Multhaupt et al.…”

“…Our results did not show any diploid DNA histogram in cases with LSIL or HSIL. However, varying percentages of diploidy and/or aneuploidy have been reported in cervical preneoplastic lesions 6–8,10,21–28 . All 17 cases of CIN3 reported by Walsh et al.…”

“…Image cytometric DNA analysis, by contrast, is an objective method with high interobserver reproducibility 4,5,33 . Based on our findings and the previous studies, a great majority of cervical preneoplastic cells (koilocytes or dysplastic cells) are non‐diploid, 21–23 whereas normal cervical mucosa, including areas adjacent to CIN3, is uniformly diploid 36,37 . Therefore, it is reasonable to consider DNA image cytometry as an appropriate technique for population‐based screening.…”

Ploidy profile of morphologically normal squamous epithelium adjacent to high grade cervical intraepithelial neoplasia

“…In addition, they have also demonstrated the presence of an abnormal ploidy profile in morphologically normal mucosa adjacent to high grade, low stage bladder tumours and found it to have independent prognostic significance 4 . In the uterine cervix the presence of aneuploidy in CIN correlates extremely well with grade, being present in up to 75% to 100% of patients with high grade lesions 5 . If ectocervical squamous epithelial aneuploidy could be shown to precede the development of a histomorpho‐ logical abnormality in CIN 2 and 3 lesions, this feature might be useful in identifying individuals who require close follow up or additional treatment of their high grade CIN.…”

Utility of Residual AutoCyte Cervical Cytology Samples for Image Analysis

“…9 A particularly useful ancillary study in cervical cytology would be DNA profiling, as performed earlier. 13,18,20,26 The process of destaining these conventional smears and restaining with the Feulgenthionin procedure, however, leads to unpredictable losses in stainable content according to some investigators. 1 Other investigators have resorted to disaggregation of nuclei from subsequent cervical biopsies in order to investigate DNA ploidy of cervical lesions.…”