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Nuclear DNA content analysis by static cytometry in cervical intraepithelial lesions using retrospective series of
N K Shirata1, N S Gomes, E A Garcia
1Istituto Adolfo Lutz, Divisione di Patologia, Avenida Dr. Arnaldo, 355 - Cerqueira Cezar. 01246-902, San Paulo, SP, Brasile.
Aims:
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.
Main 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.
Conclusion:
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.