Diagnostic Cytopathology

We have assessed the immunoc).tocliei::ic;II staining of Ki-67 nuclcar antigcn in 5 1 specimens o l uterine cervix using tlie nionocloiial antibody MIB-1 which is suitable for paraflin sections. In norm! cervical epitlielia Ki-67 inlmunostaining was conlincd to parabasal nuclei. I n biopsies showing chronic inflarmnation but no dysplasia Ki-67 imniunoslaining estended to the basal one-third of the epithelium will1 occasional positive nuclei in Uie middle one-third. A siriiilar appearance to ccrvicitis was found in mild dysplasia (CIN I); in 8 out of 11 biopsies Ki-67 inlrntlrlostairiing was seen in the basal one-third only, while in 3 cases some positive nuclei occurred in the iiiiddle one-lhird. By contrast, in all 10 biopsies judged to be dispIaying moderate dysplasia (CN 11) many Ki-67 positive%u& were found throughout the middle onetliird of the epilheliuni and in 8 out of 10 positivie nuclei occurred in tlie superficial one-third. All 10 biopsics showing severe dysplasia (CIN 111) were Ki-67 positive throughout. These findings suggest that in tlie contest of cell proliferation iiiost cases of cewical intraepilhelial neoplasia fall into a low or a high grade and that intermediate grades are uncommon. p6 Ki-67mtjnen eXp ression in evfolhted mtklial Cells in patimts 00 bladder cancer follcu UQ F.I. Y a m s , T. U r n : G.M. Reynolds, R. %Asby,* D.M.A. Wallace: J.A. Y q . D e p r m t of Patblcgy, UniversiCy of B i r m i r @ m k km-tnmt of U r o l o g ~ ' , Cuep-q Elizacetk Hosoiml Antibody t o Ki-67 a n t i g e n r e a c t w i t h a n u c l e a r a n t i g e n e x p r e s s e d d u r i n g a l l phases of t h e c e l l c y c l e a p p a r t f rom G O . The c y t o l o g i c a l assessmenL o f u r o t h e l i a l ce l l s is n o t o r i o u s l y d i f f i c u l t a s e x i o l i a t e d low g r a d e t r a n s i t i o n a l carcinoma ce l l s show l i t t l e c y t o l o g i c a l d i f f e r e n c e from e x f o l i a t e d no rma l o r r e a c t i v e i i r o t h e l i a l c e l l s . We o b t a i n e d 9 0 r e s i d u a l u r i n e specimens d u r i n g c y s t o s c o p y a t a b l a d d e r c a n c e r f o l l o w up c l i n i c . C y t o l o g i c a l smears were p r e p a r e d w i t h i n a few h o u r s , f i x e d i n a c e t o n e / m e t h a n o l and s t o r e d a t -70 C . Immunocytochemical s t a i n i n g v i t h MIB 1 (0505 immunotech, M a r s e i l l e , France) an a n t i b o d y t o Ki-67 a n c i g e n was c a r r i e d o u t on t h e f o l l o w up spec imens a long w i t h u r i n e and t i s s u e c o n t r o l s . The c y t o l o g y s l i d e s were g r a d e d by t h e number of c e l l s p o s i t i v e f o r Ki-67 a n t i g e n and g r a d e d as less t h a n 1%. 1-4J and more t h a n 5%. Comparison w i t h c y t o s c o p i c and c y t o l o g i c a l f i n d i n g s show t h a t a l l 10 cases w i t h 5% o r more of t h e u r o t h e l i a l c e l l s p o s i t i v e f o r Ki-67 a n t i g e n were p o s i t i v e o r s u s p i c i o u s on c y s t o s c o p y o r u r i n e cy to logy . E i g h t of t h e 26 c a s e s w i t h 1-4% of u r o t h e l i a l p o s i t i v e were s u s p i c i o u s o r p o s i t i v e on cys toscopy and c y t o l o g y . T h i r t e e n of 5 4 c a s e s w i t h less t h a n 1% p o s i t i v e were s u s p i c i o u s o r p o s i t i v e on c y s t o s c o p y o r c y t o l o g y . P r e v i o u s h i s t o l o g i c a l s t u d i e s on t r a n s i t i o n a l c e l l carc inoma (TCC) h a v e shown s i g n i f i c a n t o v e r l a p between normal uro the l ium and low g r a d e TCC b u t a s i g n i f i c a n t i n c r e a s e i n t h e Ki-67 i n d e x i n g r a d e 3 t u n o u r s and w i t h i n v a s i o n . Ki-67 E x p r e s s i o n i n e x f o l i a t e d u r o t h e l i a l neoplasms i s c o m p l i c a t e d by t h e p r e s e n c e of non m a l i g n a n t u r o t h e l i a l c e l l s bu t i n o u r s t u d y of TCC f o l l o w up c a s e s a Ki-67 i n d e x of 5% o r more c o r r e l a t e s well w i t h r e c u r r e n t TCC. 14 British Society for Clinical Cytology Resected + tumour block p7 Comparison of the proliferation indices, Ki-67 and S-phase fraction in fine needle aspirates from breast carcinomas. S.A. Bunon[, J.C. Titley2, P.A. Trott3 & M. Dowsettl [Academic Biochemistry, CRC Centre for Cancer Chernotherapeutics, Cytology, Royal Marsden Hospital & Institute of Cancer Research, Pathological investigation of proliferation is important for prognostic and biological studies. Measurements of mitotic index, S-phase fraction (SPF), PCNA (cyclin) and Ki-67 staining are established markers of proliferation. Measurement by (SPF) however, is generally regarded as the 'gold standard' and as such has been widely used in prognostic evaluation in the United Statcs. This technique is usually performed on digests of tissue biopsies. The first aim of our studies was to evaluate the reliability and reproducibility of SPF measurements on tine needle aspirates (FNA's) from breast carcinomas. In a group of 17 primary breast cancer patients, repeat cytological samples were taken from the m e turnour with a I-week interval without intervening treatment. In 9 I 17 cases a value for SPF was obtained from both samples. On linear regression analysis the correlation coefficient was 0.95 between recorded results, indicating a high reproducibility for this technique. between this measurement and the more widely available technique of Ki-67 staining. SPF measurements from FNA's were compared with Ki67 staining, measured by bmmunohistochernistry (using the MIB-1 antibody) from paraffin sections from the same lumour. A group of 19 patients with primary breast cancer were studied. FNA's were obtained at the time of surgical excision.The correlation coefficient was 0.80 (F 0.001) .Using the same cohort of patients, Ki-67 staining measured irnmunocytochemically from FNA cytospins was compared with SPF measurements from the same FNA. The correlation coefficient was 0.77 We conclude that S-phase and Ki-67 measurements (either by immunohistochernistry or /mmunocytochemistry) appear to be closely related markers of proliferation which are readily measurable in cytological samples. We are now taking advantage of this by making repeated measurements in patients with breast cancer during their treatment with a number of endocrine agents to compare their effects on prolifeeration.(Supported by the BSCC). Fulharn Rd, London? Downs Rd, Sutton. S-phase measurements require very expensive equipment. We therefore assessed the relationship