Author/Editor     Srebotnik-Kirbiš, Irena
Title     Imunokemično odkrivanje antigena PCNA in Ki-67 na celičnih vzorcih raka dojke
Type     monografija
Place     Ljubljana
Publisher     Medicinska fakulteta
Publication year     1998
Volume     str. 62
Language     slo
Abstract     Background: Assessment of breast cancer proliferative activity before surgery could help in planning effective individual treatment protocols, particulary in planning neo-adjuvant chemoterapy. In the present study, we are aiming to find out whether it is possible to immunochemically assess the proliferative activity of tumor on fine needle aspirates before operation. The preparation and fixation of the cell samples of immunochemical methods have not been standarized and, in addition to that, they differ from the preparation of tissue samples. We have therefore tested the effect of some fixatives on immunochemical detection of two most important proliferative antigens, PCNA and Ki-67, in breast cancer cell samples. Out particular attention has been paid to detection of these antigens in diagnostic Papanicolaou stained fine needle aspirates. Materials and Methods: The cytospins of human breast cancer cell line MCF-7 were prepared. Part of them were fixed in Delaunay's fixative or its modification and stained according to Papanicolaou. Others were fixed by Mercofix, methanol, acetone or glutaraldehyde and kept in methanol. Immunochemical detection of PCNA and Ki-67 antigens was carried out by avidin-biotin peroxidase method with PC10 and MIB-1 monoclonal antibodies, respectively. The percentage of PC10 and MIB-1 positive cells was evaluated semiquantitatively and was compared to the S phase fraction calculated from MCF-7 flow cytometric DNA histogram. Results: On the samples stained by Papanicolaou's procedure antigen Ki-67 could be immunochemically detected only after microwave pretreatment. Pretreatment with microwaves was not required for samples fixed in methanol. All other fixation methods had a negative effect on the Ki-67 antigen detection.(Abstract truncated at 2000 characteres.)
Descriptors     BREAST NEOPLASMS