Novel strategy to diagnose and grade hepatocellular carcinoma

Shahida Parveen, Riffat Mehboob, Asad Aslam Khan, Mulazim Hussain Bukhari, Fridoon Jawad Ahmad

Center for Research in Molecular Medicine, University of Lahore, Lahore, Pakistan.
Department of Pathology, King Edward Medical University, Lahore, Pakistan.
Department of Pathology, King Edward Medical University, Lahore, Pakistan;.

DOI: 10.4236/ns.2013.51004                

Abstract

Hepatocellular carcinoma (HCC) is among the most common malignancies worldwide, particularly in South and South East Asia. Unfortunately due to lack of appropriate facilities and awareness only limited information is available about its early diagnosis. Aim of the present study was to determine the efficacy of p53 by immunohistochemistry and Argyrophilicnucleolar organizer regions (AgNORs) in diagnosis of HCC and cirrhosis of liver. A total of 100 liver biopsies were studied, it included 20 cases of HCC, 60 cases of cirrhosis of the liver and 20 cases of normal liver from autopsy specimens as a control. Out of 20 cases of HCC, 15 were positive for p53 stain and 5 were negative. None of the 60 cases of cirrhosis or 20 with normal histology revealed p53 expression. A statistically significant (p < 0.001) difference was observed between mean AgNOR counts of normal (1.57 +/- 0.13), cirrhotic (4.70 +/- 0.66) and HCC tissues (14.96 +/- 1.18). In contrast the mean AgNOR count of biopsies with alcoholic cirrhosis (1.57 +/- 1.62) was significantly less (p < 0.001) than post-hepatitic cirrhosis and was similar to that of normal liver tissue. AgNORs differentiates post-hepatitic and alcoholic cirrhosis. HCV and HBV were found to be the main causative agents in HCC and Cirrhosis of liver. Mean age of HCC patients was slightly higher than liver cirrhosis patients. It is concluded that p53 and AgNORs can act as a good adjuvant to histology in diagnosing liver diseases. It helps in differentiation from well differentiated to moderately and to poorly differentiated HCC.

Keywords

P53; AgNOR; Hepatocellular Carcinoma; Cirrhosis; Immunohistochemistry

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Brugieres, L., Branchereau, S. and Laithier, V. (2012) Paediatric malignant liver tumours. Bulletin du Cancer, 99, 219-228.
[2]	Salhab, M. and Canelo, R. (2011) An overview of evidence-based management of hepatocellular carcinoma: A meta-analysis. Journal of Cancer Research and Therapeutics, 7, 463-475. doi:10.4103/0973-1482.92023
[3]	Michielsen, P.P., Francque, S.M. and van Dongen, J.L. (2005) Viral hepatitis and hepatocellular carcinoma. World Journal of Surgical Oncology, 3, 27. doi:10.1186/1477-7819-3-27
[4]	Parkin, D.M., et al. (2005) Global cancer statistics, 2002. CA: A Cancer Journal for Clinicians, 55, 74-108. doi:10.3322/canjclin.55.2.74
[5]	Farrell, G.C., Yuen, M.F., Amarapurkar, D.N., Chutaputti, A., Fan, J.G., Hou, J.L., Han, K.H., Kao, J.H., Lim, S.G., Mohamed, R., Sollano, J. and Ueno, Y. (2010) Prevention of hepatocellular carcinoma in the Asia-Pacific region: Consensus statements. Journal of Gastroenterology and Hepatology, 25, 657-663.
[6]	Altekruse, S.F., McGlynn, K.A. and Reichman, M.E. (2009) Hepatocellular carcinoma incidence, mortality, and survival trends in the United States from 1975 to 2005. Journal of Clinical Oncology, 27, 1485-1491. doi:10.1200/JCO.2008.20.7753
[7]	Capocaccia, R., et al. (2007) Hepatocellular carcinoma: Trends of incidence and survival in Europe and the United States at the end of the 20th century. American Journal of Gastroenterology, 102, 1661-1670.
[8]	Baatarkhuu, O., et al. (2011) Current situation of hepatocellular carcinoma in Mongolia. Oncology, 81, 148-151. doi:10.1159/000333278
[9]	Oyunsuren, T., et al. (2006) Hepatocellular carcinoma and its early detection by AFP testing in Mongolia. Asian Pacific Journal of Cancer Prevention, 7, 460-462.
[10]	Yuen, M.F., Hou, J.L. and Chutaputti, A. (2009) Hepatocellular carcinoma in the Asia pacific region. Journal of Gastroenterology and Hepatology, 24, 346-353. doi:10.1111/j.1440-1746.2009.05784.x
[11]	Marrero, C.R. and Marrero, J.A. (2007) Viral hepatitis and hepatocellular carcinoma. Archives of Medical Research, 38, 612-620. doi:10.1016/j.arcmed.2006.09.004
[12]	Nguyen, V.T., Law, M.G. and Dore, G.J. (2009) Hepatitis B-related hepatocellular carcinoma: Epidemiological characteristics and disease burden. Journal of Viral Hepatitis, 16, 453-463. doi:10.1111/j.1365-2893.2009.01117.x
[13]	Tarantino, G. (2007) Utility of sonography in diagnosing hepatocellular carcinoma. American Journal of Roentgenology, 188, 494-495.
[14]	Soresi, M., et al. (2003) Usefulness of alpha-fetoprotein in the diagnosis of hepatocellular carcinoma. Anticancer Research, 23, 1747-1753.
[15]	Giardina, M.G., et al. (1998) Serum alpha-L-fucosidase activity and early detection of hepatocellular carcinoma: A prospective study of patients with cirrhosis. Cancer, 83, 2468-2474. doi:10.1002/(SICI)1097-0142(19981215)83:12<2468::AID-CNCR9>3.0.CO;2-Y
[16]	Ansari, N.A. and Derias, N.W. (1997) Fine needle aspiration cytology. Journal of Clinical Pathology, 50, 541-543. doi:10.1136/jcp.50.7.541
[17]	Wee, A. (2005) Fine needle aspiration biopsy of the liver: Algorithmic approach and current issues in the diagnosis of hepatocellular carcinoma. Cytojournal, 2, 7. doi:10.1186/1742-6413-2-7
[18]	Livni, N., et al. (1995) p53 expression in patients with cirrhosis with and without hepatocellular carcinoma. Cancer, 75, 2420-2426. doi:10.1002/1097-0142(19950515)75:10<2420::AID-CNCR2820751006>3.0.CO;2-6
[19]	Oiwa, H., et al. (1995) Growth pattern and p53 overexpression in patients with early gastric cancer. Cancer, 75, 1454-1459. doi:10.1002/1097-0142(19950315)75:6+<1454::AID-CNCR2820751511>3.0.CO;2-G
[20]	Crocker, J. (1990) AgNORs and follicular lymphomas. Journal of Clinical Pathology, 43, 964-965. doi:10.1136/jcp.43.11.964
[21]	Nayak, S.K., Panesar, P.S. and Kumar, H. (2011) Nongenotoxic p53-activators and their significance as antitumor therapy of future. Current Medicinal Chemistry, 18, 1038-1049. doi:10.2174/092986711794940833
[22]	Scheistroen, M., et al. (1999) p53 protein expression in squamous cell carcinoma of the vulva. Cancer, 85, 1133-1138. doi:10.1002/(SICI)1097-0142(19990301)85:5<1133::AID-CNCR17>3.0.CO;2-T
[23]	Cheng, L., et al. (1999) p53 protein overexpression is associated with increased cell proliferation in patients with locally recurrent prostate carcinoma after radiation therapy. Cancer, 85, 1293-1299. doi:10.1002/(SICI)1097-0142(19990315)85:6<1293::AID-CNCR11>3.0.CO;2-O
[24]	Chen, F., Wang, W. and El-Deiry, W.S. (2010) Current strategies to target p53 in cancer. Biochemical Pharmacology, 80, 724-730. doi:10.1016/j.bcp.2010.04.031
[25]	Garcia, J.F., et al. (1997) p53 expression in CMV-infected cells: Association with the alternative expression of the p53 transactivated genes p21/WAF1 and MDM2. Histopathology, 30, 120-125. doi:10.1046/j.1365-2559.1997.d01-577.x
[26]	Sivridis, E. and Sims, B. (1990) Nucleolar organiser regions: New prognostic variable in breast carcinomas. Journal of Clinical Pathology, 43, 390-392. doi:10.1136/jcp.43.5.390
[27]	Smith, P.J., et al. (1988) The effect of a series of fixatives on the AgNOR technique. Journal of Pathology, 155, 109-112. doi:10.1002/path.1711550206
[28]	Beresford, M.J., Wilson, G.D. and Makris, A. (2006) Measuring proliferation in breast cancer: Practicalities and applications. Breast Cancer Research, 8, 216. doi:10.1186/bcr1618
[29]	Karki, S., Jha, A. and Sayami, G. (2012) The role of argyrophilic nucleolar organizer region (AgNoR) study in cytological evalution of fluids, especially for detection of malignancy. Kathmandu University Medical Journal (KUMJ), 10, 44-47.
[30]	Siddiqui, M.S., et al. (1999) Assessment of nucleolar organizer regions (NORs) in proliferative conditions of the liver. Pathology Research and Practice, 195, 421-426. doi:10.1016/S0344-0338(99)80016-8
[31]	WMA, (2010) World Medical Association Declaration of Helsinki: Ethical principles for medical research involveing human subjects. http://www.wma.net/e/policy/b3.htm.
[32]	Trere, D. (2000) AgNOR staining and quantification. Micron, 31, 127-131. doi:10.1016/S0968-4328(99)00069-4
[33]	Crocker, J. (1996) Molecular and biochemical aspects of interphase nucleolar organiser regions. Journal of Clinical Pathology, 49, M8-M11. doi:10.1136/mp.49.1.M8
[34]	Chiu, K.Y., Loke, S.L. and Wong, K.K. (1989) Improved silver technique for showing nucleolar organiser regions in paraffin wax sections. Journal of Clinical Pathology, 42, 992-994. doi:10.1136/jcp.42.9.992
[35]	Chiaramonte, M., et al. (1999) Rate of incidence of heaptocellular carcinoma in patients with compensated viral cirrhosis. Cancer, 85, 2132-2137. doi:10.1002/(SICI)1097-0142(19990515)85:10<2132::AID-CNCR6>3.0.CO;2-H
[36]	Roa, I., et al. (1997) p53 tumour suppressor gene protein expression in early and advanced gallbladder carcinoma. Histopathology, 31, 226-230. doi:10.1046/j.1365-2559.1997.2420850.x
[37]	Hogmo, A., et al. (1998) Nuclear DNA content and p53 overexpression in stage I squamous cell carcinoma of the tongue compared with advanced tongue carcinomas. Molecular Pathology, 51, 268-272. doi:10.1136/mp.51.5.268
[38]	Yaziji, H., et al. (1996) Role of p53 immunohistochemistry in differentiating reactive gliosis from malignant astrocytic lesions. American Journal of Surgical Pathology, 20, 1086-1090. doi:10.1097/00000478-199609000-00006
[39]	Ojanguren, I., et al. (1995) p53 immunoreactivity in heaptocellular adenoma, focal nodular hyperplasia, cirrhosis and hepatocellular carcinoma. Histopathology, 26, 63-68. doi:10.1111/j.1365-2559.1995.tb00622.x
[40]	Raedle, J., et al. (1995) Alpha-fetoprotein and p53 autoantibodies in patients with chronic hepatitis C. Digestive Diseases and Sciences, 40, 2587-2594. doi:10.1007/BF02220446
[41]	Ojanguren, I., et al. (1996) p53 immunoreaction in heaptocellular carcinoma and its relationship to etiologic factors. A fine needle aspiration study. Acta Cytologica, 40, 1148-1153. doi:10.1159/000333973
[42]	Wee, A., et al. (1994) Fine needle aspiration biopsy of hepatocellular carcinoma. Diagnostic dilemma at the ends of the spectrum. Acta Cytologica, 38, 347-354.
[43]	Darnton, S.J. and Jenner, K. (1998) Microwave irradiation lowers immunohistological detection thresholds for p53 protein in squamous epithelium from non-neoplastic oesophagus. The Journal of Pathology, 185, 334-335. doi:10.1002/(SICI)1096-9896(199807)185:3<334::AID-PATH36>3.0.CO;2-0
[44]	Iakova, P., Timchenko, L. and Timchenko, N.A. (2011) Intracellular signaling and hepatocellular carcinoma. Seminars in Cancer Biology, 21, 28-34. doi:10.1016/j.semcancer.2010.09.001
[45]	Guan, Y.S., et al. (2007) p53 gene in treatment of hepatic carcinoma: status quo. World Journal of Gastroenterology, 13, 985-992.
[46]	Hsia, C.C., et al. (2000) Correlation of immunohistochemical staining and mutations of p53 in human hepatocellular carcinoma. Oncology Reports, 7, 353-356.
[47]	Derenzini, M., et al. (1993) Is high AgNOR quantity in hepatocytes associated with increased risk of hepatocellular carcinoma in chronic liver disease? Journal of Clinical Pathology, 46, 727-729. doi:10.1136/jcp.46.8.727
[48]	Aubele, M., Auer, G. and Jutting, U. (1994) Prognostic value of AgNORs in breast cancer. Zentralblatt für Pathologie, 140, 55-60.
[49]	Crocker, J. and McGovern, J. (1988) Nucleolar organiser regions in normal, cirrhotic, and carcinomatous livers. Journal of Clinical Pathology, 41, 1044-1048. doi:10.1136/jcp.41.10.1044
[50]	Dervan, P.A., et al. (1989) Breast carcinoma kinetics. Argyrophilic nucleolar organizer region counts correlate with Ki67 scores. American Journal of Clinical Pathology, 92, 401-407.
[51]	Raymond, W.A. and Leong, A.S. (1989) Nucleolar organizer regions relate to growth fractions in human breast carcinoma. Human Pathology, 20, 741-746. doi:10.1016/0046-8177(89)90066-X
[52]	Ascoli, V., et al. (1990) Nucleolar organizer regions in the fine needle aspirates of lung tumours. Cytopathology, 1, 277-286. doi:10.1111/j.1365-2303.1990.tb00361.x
[53]	Allen, J.P. and Gallimore, A.P. (1992) Nucleolar organizer regions in benign and malignant glandular lesions of the cervix. The Journal of Pathology, 166, 153-156. doi:10.1002/path.1711660211
[54]	Pich, A., Margaria, E. and Chiusa, L. (2002) Significance of the AgNOR in tumor pathology. Pathologica, 94, 2-9.
[55]	Yu, C.C., et al. (1992) A comparison of proliferating cell nuclear antigen (PCNA) immunostaining, nucleolar organizer region (AgNOR) staining, and histological grading in gastrointestinal stromal tumours. The Journal of Pathology, 166, 147-152. doi:10.1002/path.1711660210
[56]	Shiro, T., et al. (1993) A correlation of argyrophilic nucleolar organizer regions with stages of hepatocellular carcinoma. Cancer, 71, 44-49. doi:10.1002/1097-0142(19930101)71:1<44::AID-CNCR2820710108>3.0.CO;2-L