Summary Recent preclinical and clinical data suggest that TP53 status and TP53 mutations may be important in determining tumour aggressiveness and therapy response. In this study we investigate the feasibility of a structural and quantitative analysis of TP53 on fineneedle aspiration (FNA) material obtained from 31 consecutive female patients with breast carcinoma, enrolled in a primary chemotherapy protocol. Tumours were screened for p53 protein overexpression and TP53 mutations (exons 5-8) using immunocytochemistry, polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and DNA sequencing analyses, and finally using fluorescence in situ hybridization (FISH) analysis. Positive nuclear staining was identified in six cases whereas mutations were detected in nine. Although the immunoreactive pattern fitted fully with the characterized TP53 mutation type, the considerable number of null p53 mutations (i.e. four) coupled with the lack of information regarding the localization of TP53 mutations make immunocytochemistry an inadequate indicator of TP53 function deregulation. Combining molecular and FISH analyses, we detected three cases with TP53 deletion and one case with deletion and mutation. Finally, DNA static-image analysis performed on 29 cases showed aneuploidy in 26 cases, which included all TP53-mutated cases. The present results show that FNA may assist clinical decisions by allowing the evaluation of a variety of biological parameters relevant for prognosis and treatment planning.Keywords: fine-needle aspiration; breast carcinoma; TP53 analysis; fluorescence in situ hybridization analysis; DNA content analysisThe fine-needle aspiration (FNA) technique represents one of the most effective and versatile methodologies that contribute to the diagnosis and the management of breast cancer. Over recent years, the field of FNA application has expanded, and currently FNA spans the diagnostic assessment of palpable and non-palpable nodules and contributes to management decisions at surgical and medical levels (Masood et al, 1990;Layfield, 1992). In fact, FNA has largely replaced frozen sections and, in cases candidated to primary chemotherapy, provides hormonal receptor assessment as well as other useful pathobiological parameters with an efficiency that equals that of histology (Thomas et al, 1990;Dowell et al, 1994;Leong et al, 1996).Recent clinical evidence supports a critical role of TP53 status in providing prognostic information (Kovach et al, 1996;Sjogren et al, 1996). Preclinical (Lowe et al, 1993 Wahl et al, 1996) and clinical data (Bergh et al, 1995; Aas et al, 1996;Fricker, 1996;Sjogren et al, 1996) suggest that TP53 status and TP53 mutation types may be important not only in determining tumour aggressiveness but also in the response to therapy. In fact, there is increasing evidence that tumours lacking normal TP53 function are clinically more aggressive as they acquire a selective growth advantage becoming more resistant to ionizing radiations and some widely used anti-cancer drugs...