Initial liver biopsies from 28 patients with cirrhosis, included in a follow-up program for hepatocellular carcinoma (HCC) surveillance, were investigated to verify whether cell proliferative activity was associated with the development of HCC. Cirrhosis was related to HBV in 15 cases, HCV in 11 and HBV/HCV in 2. On admission, no liver dysplasia was evident. During the follow-up period, 9 patients (32.1%) developed HCC. Sections, pretreated for 20 min at 120 °C in pH 6 citrate buffer, were stained with 50% silver nitrate at 37 °C for 14 min. AgNOR protein area from 200 cells in each case was measured with an automated image analyser, using the program AgNOR (Immagini & Computer, Rho, Italy). Cell proliferative activity was defined as the percentage of hepatocytes with AgNOR area greater than 7m2 (AgNOR-PI, proliferation index ) (Trerè et al. Eur J Histochem 40, 351, 1996). The mean AgNOR protein area for the whole series was 3.42m2. It was greater in patients who developed HCC (3.68m2) than in those who did not undergo neoplastic transformation (3.29m2), and in HBV/HCV related cirrhosis (4.58) than in cirrhosis related to HBV (3.38m2) or HCV (3.25m2) alone. No association was found with flogosis, fibrosis or steatosis. Neoplastic transformation occurred in four of 5 patients (80%) with AgNOR-PI greater than 5, but only in 5 out 23 (21.7%) patients with a lower AgNOR-PI (Fisher's exact test: 0.025). With the limitation due to the small number of cases, our results indicate that AgNOR-PI, could be regarded as a reproducible predictor of HCC development in patients with liver cirrhosis.
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