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. 1998 Apr;51(2):96–101. doi: 10.1136/mp.51.2.96

Hepatocyte proliferation rate is a powerful parameter for predicting hepatocellular carcinoma development in liver cirrhosis.

M Borzio 1, D Trerè 1, F Borzio 1, A R Ferrari 1, S Bruno 1, M Roncalli 1, G Colloredo 1, G Leandro 1, F Oliveri 1, M Derenzini 1
PMCID: PMC395617  PMID: 9713593

Abstract

AIMS: A sound predictive test is lacking for the identification of cirrhotic patients at high risk of developing hepatocellular carcinoma. The present study evaluates the measurement of hepatocyte expression of silver stained nucleolar organiser region (AgNOR) proteins as a risk factor for the development of hepatocellular carcinoma in cirrhosis. METHODS: Liver biopsies from 176 cirrhotic patients included in a follow up surveillance programme for hepatocellular carcinoma development were evaluated prospectively for hepatocyte AgNOR protein quantity. The follow up programme consisted of clinical and biochemical assessment every three months, and ultrasound scanning and serum alpha-fetoprotein (alpha FP) assessment every six months. Histological sections from the needle biopsies performed at enrollment were stained selectively for AgNOR proteins and the percentage of hepatocytes with an AgNOR protein area > or = 7 micron 2, indicative of a proliferative state (AgNOR proliferation index (AgNOR-PI)), was measured. RESULTS: During the mean (SD) follow up time of 65.5 (36.29) months (range, 12-143; median, 67), hepatocellular carcinoma was diagnosed in 48 of 176 patients (27.3%). The AgNOR-PI of the whole series ranged from 0% to 5% (median, 0.9%), and was significantly higher in patients with liver cell dysplasia and hepatitis B surface antigen (HBsAg) positivity (p < 0.0001 and p = 0.0002, respectively). The 176 patients were divided into two groups according to their AgNOR-PI scores; a cut off value of 2.5% defined by the receiver operating characteristic curve and the Youden index was used. Forty two patients were included in the high AgNOR-PI (< 2.5%) group, and 134 patients the low AgNOR-PI (< 2.5%) group. In the high AgNOR-PI group, 25 of 42 patients developed hepatocellular carcinoma, in contrast to only 23 of 134 patients (17.2%) in the group with a low AgNOR-PI (p < 0.0001). Hepatocellular carcinoma development was also significantly more frequent in patients with liver cell dysplasia and HBsAg positivity. Multivariate analysis using AgNOR-PI, liver cell dysplasia, HBsAg positivity, and hepatitis C virus (HCV) infection as covariates demonstrated that the AgNOR-PI parameter was the only significant predictor of hepatocellular carcinoma development. CONCLUSIONS: These results demonstrate that a high hepatocyte proliferation rate is a major risk factor for hepatocellular carcinoma development in the cirrhotic liver. Therefore, the evaluation of the hepatocyte proliferation rate is very important to identify patients requiring a more strict follow up programme for early diagnosis of hepatocellular carcinoma.

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Selected References

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  1. Anthony P. P., Ishak K. G., Nayak N. C., Poulsen H. E., Scheuer P. J., Sobin L. H. The morphology of cirrhosis. Recommendations on definition, nomenclature, and classification by a working group sponsored by the World Health Organization. J Clin Pathol. 1978 May;31(5):395–414. doi: 10.1136/jcp.31.5.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anthony P. P., Vogel C. L., Barker L. F. Liver cell dysplasia: a premalignant condition. J Clin Pathol. 1973 Mar;26(3):217–223. [PMC free article] [PubMed] [Google Scholar]
  3. Ballardini G., Groff P., Zoli M., Bianchi G., Giostra F., Francesconi R., Lenzi M., Zauli D., Cassani F., Bianchi F. Increased risk of hepatocellular carcinoma development in patients with cirrhosis and with high hepatocellular proliferation. J Hepatol. 1994 Feb;20(2):218–222. doi: 10.1016/s0168-8278(05)80061-3. [DOI] [PubMed] [Google Scholar]
  4. Borzio M., Bruno S., Roncalli M., Mels G. C., Ramella G., Borzio F., Leandro G., Servida E., Podda M. Liver cell dysplasia is a major risk factor for hepatocellular carcinoma in cirrhosis: a prospective study. Gastroenterology. 1995 Mar;108(3):812–817. doi: 10.1016/0016-5085(95)90455-7. [DOI] [PubMed] [Google Scholar]
  5. Colombo M., de Franchis R., Del Ninno E., Sangiovanni A., De Fazio C., Tommasini M., Donato M. F., Piva A., Di Carlo V., Dioguardi N. Hepatocellular carcinoma in Italian patients with cirrhosis. N Engl J Med. 1991 Sep 5;325(10):675–680. doi: 10.1056/NEJM199109053251002. [DOI] [PubMed] [Google Scholar]
  6. Crocker J., McGovern J. Nucleolar organiser regions in normal, cirrhotic, and carcinomatous livers. J Clin Pathol. 1988 Oct;41(10):1044–1048. doi: 10.1136/jcp.41.10.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Derenzini M., Ploton D. Interphase nucleolar organizer regions in cancer cells. Int Rev Exp Pathol. 1991;32:149–192. doi: 10.1016/b978-0-12-364932-4.50008-3. [DOI] [PubMed] [Google Scholar]
  8. Derenzini M., Trerè D., Oliveri F., David E., Colombatto P., Bonino F., Brunetto M. R. Is high AgNOR quantity in hepatocytes associated with increased risk of hepatocellular carcinoma in chronic liver disease? J Clin Pathol. 1993 Aug;46(8):727–729. doi: 10.1136/jcp.46.8.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ganne-Carrié N., Chastang C., Chapel F., Munz C., Pateron D., Sibony M., Dény P., Trinchet J. C., Callard P., Guettier C. Predictive score for the development of hepatocellular carcinoma and additional value of liver large cell dysplasia in Western patients with cirrhosis. Hepatology. 1996 May;23(5):1112–1118. doi: 10.1002/hep.510230527. [DOI] [PubMed] [Google Scholar]
  10. Huang S. N., Chisari F. V. Strong, sustained hepatocellular proliferation precedes hepatocarcinogenesis in hepatitis B surface antigen transgenic mice. Hepatology. 1995 Mar;21(3):620–626. [PubMed] [Google Scholar]
  11. Kuo G., Choo Q. L., Alter H. J., Gitnick G. L., Redeker A. G., Purcell R. H., Miyamura T., Dienstag J. L., Alter M. J., Stevens C. E. An assay for circulating antibodies to a major etiologic virus of human non-A, non-B hepatitis. Science. 1989 Apr 21;244(4902):362–364. doi: 10.1126/science.2496467. [DOI] [PubMed] [Google Scholar]
  12. Mazzella G., Accogli E., Sottili S., Festi D., Orsini M., Salzetta A., Novelli V., Cipolla A., Fabbri C., Pezzoli A. Alpha interferon treatment may prevent hepatocellular carcinoma in HCV-related liver cirrhosis. J Hepatol. 1996 Feb;24(2):141–147. doi: 10.1016/s0168-8278(96)80022-5. [DOI] [PubMed] [Google Scholar]
  13. Mourad W. A., Connelly J. H., Sembera D. L., Atkinson E. N., Bruner J. M. The correlation of two argyrophilic nucleolar organizer region counting methods with bromodeoxyuridine-labeling index: a study of metastatic tumors of the brain. Hum Pathol. 1993 Feb;24(2):206–210. doi: 10.1016/0046-8177(93)90302-w. [DOI] [PubMed] [Google Scholar]
  14. Mourad W. A., Erkman-Balis B., Livingston S., Shoukri M., Cox C. E., Nicosia S. V., Rowlands D. T., Jr Argyrophilic nucleolar organizer regions in breast carcinoma. Correlation with DNA flow cytometry, histopathology, and lymph node status. Cancer. 1992 Apr 1;69(7):1739–1744. doi: 10.1002/1097-0142(19920401)69:7<1739::aid-cncr2820690715>3.0.co;2-9. [DOI] [PubMed] [Google Scholar]
  15. Munakata S., Hendricks J. B. Effect of fixation time and microwave oven heating time on retrieval of the Ki-67 antigen from paraffin-embedded tissue. J Histochem Cytochem. 1993 Aug;41(8):1241–1246. doi: 10.1177/41.8.8331288. [DOI] [PubMed] [Google Scholar]
  16. Ofner D., Bankfalvi A., Riehemann K., Bier B., Böcker W., Schmid K. W. Wet autoclave pretreatment improves the visualization of silver-stained nucleolar organizer-region-associated proteins in routinely formalin-fixed and paraffin-embedded tissues. Mod Pathol. 1994 Dec;7(9):946–950. [PubMed] [Google Scholar]
  17. Pession A., Farabegoli F., Treré D., Novello F., Montanaro L., Sperti S., Rambelli F., Derenzini M. The Ag-NOR proteins and transcription and duplication of ribosomal genes in mammalian cell nucleoli. Chromosoma. 1991 May;100(4):242–250. doi: 10.1007/BF00344158. [DOI] [PubMed] [Google Scholar]
  18. Ploton D., Menager M., Jeannesson P., Himber G., Pigeon F., Adnet J. J. Improvement in the staining and in the visualization of the argyrophilic proteins of the nucleolar organizer region at the optical level. Histochem J. 1986 Jan;18(1):5–14. doi: 10.1007/BF01676192. [DOI] [PubMed] [Google Scholar]
  19. Sirri V., Roussel P., Trerè D., Derenzini M., Hernandez-Verdun D. Amount variability of total and individual Ag-NOR proteins in cells stimulated to proliferate. J Histochem Cytochem. 1995 Sep;43(9):887–893. doi: 10.1177/43.9.7642962. [DOI] [PubMed] [Google Scholar]
  20. Tarao K., Ohkawa S., Shimizu A., Harada M., Nakamura Y., Ito Y., Tamai S., Hoshino H., Inoue T., Kanisawa M. Significance of hepatocellular proliferation in the development of hepatocellular carcinoma from anti-hepatitis C virus-positive cirrhotic patients. Cancer. 1994 Feb 15;73(4):1149–1154. doi: 10.1002/1097-0142(19940215)73:4<1149::aid-cncr2820730405>3.0.co;2-9. [DOI] [PubMed] [Google Scholar]
  21. Tarao K., Ohkawa S., Shimizu A., Harada M., Nakamura Y., Ito Y., Tamai S., Hoshino H., Okamoto N., Iimori K. The male preponderance in incidence of hepatocellular carcinoma in cirrhotic patients may depend on the higher DNA synthetic activity of cirrhotic tissue in men. Cancer. 1993 Jul 15;72(2):369–374. doi: 10.1002/1097-0142(19930715)72:2<369::aid-cncr2820720210>3.0.co;2-5. [DOI] [PubMed] [Google Scholar]
  22. Tarao K., Shimizu A., Ohkawa S., Harada M., Ito Y., Tamai S., Kuni Y., Okamoto N., Inoue T., Kanisawa M. Development of hepatocellular carcinoma associated with increases in DNA synthesis in the surrounding cirrhosis. Gastroenterology. 1992 Aug;103(2):595–600. doi: 10.1016/0016-5085(92)90852-p. [DOI] [PubMed] [Google Scholar]
  23. Torp S. H. Proliferative activity in human glioblastomas: evaluation of different Ki67 equivalent antibodies. Mol Pathol. 1997 Aug;50(4):198–200. doi: 10.1136/mp.50.4.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Trerè D., Derenzini M., Sirri V., Montanaro L., Grigioni W., Faa G., Columbano G. M., Columbano A. Qualitative and quantitative analysis of AgNOR proteins in chemically induced rat liver carcinogenesis. Hepatology. 1996 Nov;24(5):1269–1273. doi: 10.1002/hep.510240547. [DOI] [PubMed] [Google Scholar]

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