Nuclear DNA content analysis by static cytometry in cervical intraepithelial lesions using retrospective series of previously stained PAP smears.
OBJECTIVE
To evaluate the nuclear DNA content in cytological smears of uterine cervix previously stained with Papanicolaou method, focusing the potential of the method in retrospective series.


METHODS
Consecutive cases of Pap smears examined at the Adolfo Lutz Institute, a Public Health Laboratory of São Paulo State were selected. The diagnosis were: CIN 1 (n=20), CIN 2 (n=24), CIN 3 (n=20). Slides were previously stained with Papanicolaou method. The stain was removed with 5% hydrochloric alcohol-acid solution and the slides were stained with Thionin-Feulgen using a Becton & Dickinson kit. Ploidy evaluation was performed using the DNA Quantitative Measurement software 3.0 (version 8.1) from Becton & Dickinson and the CAS 200 system of image analysis. Cell ploidy was evaluated after analysis of atypical nuclei found in the selected cases. The DNA index was obtained using histograms for interpretation.


RESULTS
CIN 1 cases showed the following DNA profile: 55% of diploid, 5% of tetraploid and 40% of aneuploid. CIN 2 cases showed 45.8% of diploid, 8.3% of tetraploid and 45.8% of aneuploid cells. CIN 3 cases showed 10% diploid, 15% of tetraploid and 75% of aneuploid cells.


CONCLUSIONS
DNA analysis was useful to distinguish CIN 3 to CIN 1 and 2, but did not discriminate CIN 1 and 2 in these series. Aneuploidy was strongly associated to the CIN 3 cases.
Shirata
N K
NK
Istituto Adolfo Lutz, Divisione di Patologia, Avenida Dr. Arnaldo, 355 - Cerqueira Cezar. 01246-902, San Paulo, SP, Brasile.
Gomes
N S
NS
Garcia
E A
EA
Longatto Filho
A
A
D002467:Cell Nucleus / Q000235:genetics* / Q000473:pathology; D004396:Coloring Agents; D004273:DNA, Neoplasm / Q000032:analysis*; D005260:Female; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D007091:Image Processing, Computer-Assisted; D065006:Papanicolaou Test; D011003:Ploidies; D012189:Retrospective Studies; D012394:Rosaniline Dyes; D013194:Staining and Labeling; D002583:Uterine Cervical Neoplasms / Q000235:genetics* / Q000473:pathology; D014626:Vaginal Smears; D002578:Uterine Cervical Dysplasia / Q000235:genetics* / Q000473:pathology
D004396:Coloring Agents; D004273:DNA, Neoplasm; C007722:Feulgen stain; D012394:Rosaniline Dyes