Cyclins - a New Marker to Assess Severity of Cervical Lesions

R. Stănculescu, V. Bauşic, L. Cruceru, A. Popa, M. Russu, Z. Ceauşu, M. Ceauşu, A. Cucu, J. Marin, T. Vlădescu

Abstract: The purpose of this study is to highlight the presence of cyclins in cell cultures (CulCel) from cervical biopsies with varying degrees of lesions, using immunofluorescence with anticycline monoclonal antibodies (AcMcAC). The study included 68 samples from cases with cervical lesions (cervicitis, ASC-H, H-SIL, cervical cancer). Cultivation of explants and use of keratinocyte restraining media led to epithelial cell cultures free of contamination with fibroblasts. The obtained material was incubated with anticycline monoclonal antibodies (A, B, D1, D2, D3, E). The technique applied was indirect immunofluorescence using secondary antibodies. The interpretation of analyzed issues was based on observational criteria and was performed by three specialists in molecular and cellular biology. To provide a correlation between parameters, statistical analysis was performed using t-Student test. Cyclin A was identically expressed in the cytoplasm of the four categories of lesions, but presented significant differences in the nuclei: HPV negative cervicitis in 29% of cells, HPV positive HSIL in 42% of cells, HPV negative carcinomas in 62% of cells, HPV positive carcinomas in 71% of cells. Cyclin B in non-invasive lesions shows a moderate reaction in the dividing cells; in the presence of HPV, the response occurs in 6% of the cells in interphase. In HPV negative carcinoma, intense reaction occurs in the dividing cells and moderate reaction occurs in 21% of the interphase cells; in HPV positive carcinoma, the reaction is intense in the dividing cells and in 28% of the interphase cells. Cyclins D and E showed no significant differences between the categories of cases examined. Positive statistical correlations were observed between cyclin level in nuclei and cytoplasm, in cells with HPV negative ASC-H lesions (r = 0.75, p = 0.05), HPV positive HSIL (r = 0.77, p = 0.05), HPV negative CC - (r = 0.78, p = 0.04) and HPV positive CC (r = 0.42, p = 0.01). The presence of cyclin A in over 42% of CulCel and of cyclin B in over 21% of CulCel demonstrates their involvement as cell cycle regulatory markers in supporting the diagnosis of high grade cervical lesions/cancer.
Keywords: culture cells, cyclins, immunoflourescence, anticycline-antibodies