Case Control Study
Copyright ©The Author(s) 2016. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jul 21, 2016; 22(27): 6246-6256
Published online Jul 21, 2016. doi: 10.3748/wjg.v22.i27.6246
Immunohistochemistry panel segregates molecular types of hepatocellular carcinoma in Brazilian autopsy cases
Aloísio Felipe-Silva, Alda Wakamatsu, Cinthya dos Santos Cirqueira, Venâncio Avancini Ferreira Alves
Aloísio Felipe-Silva, LIM 14-Patologia Hepática, Faculdade de Medicina da USP, 01246-903 São Paulo SP, Brazil
Aloísio Felipe-Silva, Alda Wakamatsu, Cinthya dos Santos Cirqueira, Venâncio Avancini Ferreira Alves, Department of Pathology, Medical Research Laboratory LIM14, University of Sao Paulo School Of Medicine, 01246-903 São Paulo SP, Brazil
Author contributions: Felipe-Silva A designed and performed the research, analyzed the data, and wrote the manuscript; Wakamatsu A and dos Santos Cirqueira C contributed optimal technical support on construction of the tissue microarray and performance of the histological and immunohistochemistry reactions; and Alves VAF analyzed the data, wrote the manuscript, and supervised all the research steps.
Supported by Fundação de Amparo a Pesquisa do Estado de São Paulo, No. 08/58855-3 to Alves VAF.
Institutional review board statement: This research was approved by the Ethics Committee for Research Project Analysis (CAPPesq) of the Hospital das Clínicas of the University of Sao Paulo School Of Medicine (Protocol No. 2346-0484/08 on 06/24/08).
Conflict-of-interest statement: None of the authors has any potential or real conflict of interest related to this study or the publication of its findings. This study reports major findings of a PhD thesis by the first author performed at the University of Sao Paulo and was partially presented at the 2014 United States and Canadian Academy of Pathology (USCAP) Annual Meeting in San Diego, USA.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
Correspondence to: Aloísio Felipe-Silva, MD, PhD, LIM 14-Patologia Hepática, Faculdade de Medicina da USP, Avenida Dr. Arnaldo 455 1o. Andar Sala 1157, 01246-903 São Paulo SP, Brazil.
Telephone: +55-11-30617433 Fax: +55-11-30617234
Received: March 22, 2016
Peer-review started: March 23, 2016
First decision: April 14, 2016
Revised: May 19, 2016
Accepted: June 15, 2016
Article in press: June 15, 2016
Published online: July 21, 2016

AIM: To assess the distribution of proteins coded by genes reported as relevant for the molecular classification of hepatocellular carcinoma (HCC).

METHODS: In this retrospective cross-sectional study, the following clinicopathological data were analyzed in 80 autopsied HCC patients: sex, age, ethnicity, alcohol intake, infection with hepatitis B and/or C virus, infection with human immunodeficiency virus, prior treatment, basic and immediate causes of death, liver weight, presence of cirrhosis, number and size of nodules, gross pattern, histological grade and variants, architectural pattern, invasion of large veins, and presence and location of extrahepatic metastases. The protein products of genes known to be involved in molecular pathogenesis of HCC, including epidermal growth factor receptor (EGFR), MET, keratin 19 (K19), vimentin, beta-catenin, mechanistic target of rapamycin (mTOR), extracellular signaling-related kinase (ERK)1, ERK2, Ki67, cyclin D1, caspase 3 and p53, were detected by immunohistochemistry on tissue microarrays. The expression levels were scored and statistically assessed for correlation with HCC parameters.

RESULTS: Infection with hepatitis C virus was identified in 49% of the 80 autopsy patients, cirrhosis in 90%, advanced tumors in 95%, and extrahepatic metastases in 38%. Expression of K19, p53 and ERK1 correlated to high-grade lesions. Expression of ERK1, nuclear beta-catenin, cyclin D1 and ERK2 correlated to higher rates of cell proliferation as determined by Ki67. Expression of MET, EGFR (> 0) and caspase 3 correlated with lower histological grades. Expression of EGFR correlated to that of caspase 3, and overexpression of EGFR (≥ 200/300) was observed in low-grade tumors more frequently (grades 1 and 2: 67% vs grade 3: 27% and grade 4: 30%). Expression of ERK1 was associated with that of K19 and vimentin, whereas expression of ERK2 was associated with that of cyclin D1, MET and membrane beta-catenin. Expression of vimentin was strongly correlated with that of K19.

CONCLUSION: Expression of K19, p53, ERK1, ERK2, vimentin and nuclear beta-catenin was related to higher-grade markers, as opposed to expression/overexpression of EGFR, MET and caspase 3.

Keywords: Hepatocellular carcinoma, Epidermal growth factor receptor, Autopsy, Immunohistochemistry, Liver, Classification

Core tip: This study assessed the immunohistochemistry-detected expression of several protein products of genes known to be involved in the molecular pathogenesis of hepatocellular carcinoma (HCC) in a retrospective autopsy cohort of patients with HCC. The data showed that expression profiles of these markers may be related to different pathways underlying HCC progression and metastasis, and that the Edmondson-Steiner’s tumor grade may reflect currently recognized molecular subclasses of HCC. This cross-sectional analysis supports the strategy of translating genomic data into panels of immunohistochemical markers for risk evaluation in HCC and also reinforces the paramount importance of histological grade in this context.