Hepatocellular carcinoma: microstructure and expression features of hepatocyte marker, alphafetoprotein, cytokeratins 7 and 20
DOI:
https://doi.org/10.14739/2310-1237.2014.1.25929Keywords:
hepatocellular carcinoma, HepPar-1, AFP, СК7, СК20Abstract
Introduction.
Hepatocellular carcinoma (HCC) is the fifth most common malignant disease in men and the eighth most common in women worldwide [1]. HCC is one of the most aggressive cancers. Timely diagnostics of primary liver cancer allows to prolong and improve the quality of the patient's life. Immunohistochemical and pathological analysis of liver biopsy plays a crucial role in the differentiation of HCC from cholangiocarcinoma (CC).
HepPar-1 (Hepatocyte Specific Antigen), alpha-fetoprotein (AFP), polyclonal CEA (pCEA), monoclonal carcinoembryonic antigen (mCEA), CD10, glypican-3, factor XIIIa, alpha-1-antitrypsin and also cytokeratins (CK) 7, 8, 19, 20 are commonly used to differentiate HCC from cholangiocarcinoma (CC) [2, 3, 4]. However, expression of these markers by the cells of HCC and CC can be inhomogeneous. Furthermore the utility of each of these markers is limited by sensitivity or difficulty in interpretation.
Aim of investigation: to determine the microscopic features of HCC and study the expression of Hepatocyte Specific Antigen (HepPar-1), alpha-fetoprotein (AFP), cytokeratin 7, 20 (CK7,CK20) and area of immunopositive cells using liver biopsies from patients with hepatocellular carcinoma
Materials and methods. The complex histopathological, histochemical and immunohistochemical research was performed using liver biopsy of 53 patients with hepatocellular carcinoma. The age of patients was from 26 up to 73 years (59,6±11,32). Five liver biopsies of patients with hemangioma without any liver disease were used as samples for test-control.
Expression levels of HepPar-1, AFP, CK7 and CK20 were quantitated by photo digital morphometry at which the slides of liver cancer with appropriate immunopositive reaction were photographed by digital camera «Olympus 3040» (Japan) in microscope Axioplan 2 («Carl Zeiss», Germany) with magnification x200 in 5 fields of view and subsequently analyzed using a medical digital imaging program Image J [Rasband WS (1997-2012)]. The data was analyzed and displayed using «STATISTICA® for Windows 6.0» (StatSoft Inc., license № AXXR712D833214FAN5). To calculate the expression level of HepPar-1, alphafetoprotein (AFP), cytokeratin 7, 20 and area of immunopositive cells Image J program was used. To compare the results Pearson’s correlation was applied. P values of less than 0.05 were considered significant.
Results.
1. Expression of HepPar-1 by the cells of HCC cancer was positive in 92.45 % cases. The area of immunopositive cells was 49,35 ± 25,45%. The high level of expression of HepPar-1 was detected in 54,72% patients. The moderate level of expression of this marker was defined in 22,64% patients and low expression of HepPar-1 by the cells of hepatocellular carcinoma was detected in 1,51% patients.
2. 81,13% of patients with hepatocellular carcinoma had the cytoplasmic and nuclear expression of α-fetoprotein. The area of immunopositive cells was 49,35 ± 25,45% in average. The high level of expression of α- fetoprotein by the cells of hepatocellular carcinoma was detected in 37,74% patients. The moderate level of expression of this marker was defined in 26,42% patients and low expression of HepPar-1 was detected in 16,97% patients.
3. Cytokeratins 7 expression was noted in 37,74 % cases of HCC. Immunopositive cells were distributed in the tumor as focal accumulations and the area of immunopositive cells was 21,08±5,19%. The low expression of СК7 by the cells of hepatocellular carcinoma was detected in 22,64% patients. The moderate level of expression of this marker was defined in 10,37% patients and high level expression of СК7 was detected in 4,73% patients.
4. Cytokeratins 20 expression by the cells of hepatocellular carcinoma was positive in 30,13% cases. The area of immunopositive cells was 29,35±17,31%. The high level of expression of СК20 was detected in 19,74% patients. The moderate level of expression of this marker was defined in 4,93% patients and low expression of HepPar-1 by the cells of hepatocellular carcinoma was detected in 5,46% patients.
5. Patients with hepatocellular carcinoma have a direct weak connection between the expression level of HepPar-1 and AFP (Pearson’s coefficient is r=+0,25). There was direct medium strength connection between the expression level of AFP and СК7, AFP and by СК20 by the tumor cells (Pearson’s coefficient is r=+0,5). There was strong direct connection between the level of expression of HepPar-1 and CК7, HepPar-1 and CК7 (Pearson’s coefficient is r=+1).
Conclusion. The results from our study should be considered in the field of the differential immunohistochemical diagnostic of hepatocellular carcinoma.
References
Jelic S., Sotiropoulos G.C.( 2010) Hepatotselliuliarnyi rak: klynycheskye rekomendatsyy ESMO po dyahnostyke, lechenyiu y nabliudenyiu. S.Jelic, G.C.Sotiropoulos. V kn.: Mynymalnыe klynycheskye rekomendatsyy Evropeiskoho Obshchestva Medytsynskoi Onkolohyy (ESMO) M.: Yzd. hr. RONTs ym. N. N. Blokhyna RAMN. 436 s., 92-102.
Wang L., Vuolo M., Suhrland M.J., Schlesinger K. (2006). HepPar1, MOC, pCEA, mCEA and CD10 for distinguishing hepatocellular carcinoma vs. metastatic adenocarcinoma in liver fine needle aspirates. Acta Cytol. V 50, 257–262.
Basturk О., Farris III А.В., Adsay N. V. (2010). Immunohistology of the Pancreas, Biliary Tract and Liver. In: Diagnostic immunohistochemistry: theranostic and genomic applications. Еd. by David J. Dabbs. 3rd ed. Philadelphia: Saunders/Elsevie, 541-592.
Durnez A., Verslype C., Nevens F. ( 2006). The clinicopathological and prognostic relevance of cytokeratin 7 and 19 expression in hepatocellular carcinoma. A possible progenitor cell origin. Histopathology.Vol. 49,138-151.
Hamilton S.R., Aaltonen L.A. (2000). Pathology and genetics of tumours of the digestive system . In: World Health Organization Classification of Tumours. Lyon, France: International Agency for Research on Cancer Press, 163–166.
Hirohashi S., Ishak K.G., Kojiro M. (2000). Hepatocellular carcinoma. In: Hamilton S.R., Aaltonen L.A., Tumours of the Digestive System. Lyon, France: International Agency for Research on Cancer (IARC), 159-172.
McKenna B., Bihlmeyer Sh.( 2010). Pathology of hepatocellular carcinoma, cholangiocarcinoma and combined hepatocellular-cholangiocarcinoma. In: Primary Carcinomas of the Liver. Ed. Adviser J.E. Husband: Cambridge University Press,16-32.
Suriawinata A.A., Thung S.N. (2011).Liver pathology: an atlas and concise guide . Arief A. Suriawinata, Swan N. Thung. New York: demosMEDICAL, 260 p.
Butler S.L., Dong H., Cardona D. et al. (2008).The antigen for Hep Par 1 antibody is the urea cycle enzyme carbamoyl phosphate synthetase 1. Lab Invest. Vol. 88,78–88.
Lugli A., Tornillo L., Mirlacher M. (2004).Hepatocyte paraffin 1 expression in human normal and neoplastic tissues: tissue microarray analysis on 3,940 tissue samples. Am J Clin Pathol. vol.122, 721–727.
Shiran M.S., Isa M.R., Sherina M.S., Rampal L., Hairuszah I., Sabariah A.R. (2006). The utility of Hepatocyte Paraffin 1 antibody in the immunohistological distinction of hepatocellular carcinoma from cholangiocarcinoma and metastatic carcinoma. Malaysian J Pathol. Vol. 28 (2), 87–92.
Tuffakha M.S.A., Hychka S.H., Husky H.L. (2013) Ymmunohystokhymycheskaia dyahnostyka opukholei. –Kyev: OOO «Yntermed». 223s.
Rodyonov S. Yu., Cherkasov V. A., Maliutyna N. N., Orlov O. A. (2004) Alfa-fetoproteyn. Ekaterynburh: UrO RAN. 376s.
Kakar S., Gown A.M., Goodman Z.D., Ferrell L.D. (2007). Best practices in diagnostic immunohistochemistry: hepatocellular carcinoma versus metastatic neoplasms. Arch Pathol Lab Med. Vol. 131,1648-1654.
Burt A.D., Portmann B.C., Ferrell L.D. (2012). MacSween's Pathology of the Liver. 6th edn. Edinburgh: Churchill Livingstone/Elsevier, 1032р.
Fanni D., Nemolato S., Ganga R., Senes G., Gerosa C., Van Eyken P., Geboes K., Faa G.( 2009). Cytokeratin 20-positive hepatocellular carcinoma. European J of Histochemistry, Vol. 53(issue 4), 269-274.
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