MACC1 as a Prognostic Biomarker for Early-Stage and AFP-Normal Hepatocellular Carcinoma

Citation: Xie C, Wu J, Yun J, Lai J, Yuan Y, et al. ( MACC1 as a Prognostic Biomarker for Early-Stage and AFP-Normal Hepatocellular Carcinoma Chan Xie 0 Jueheng Wu 0 Jingping Yun 0 Jiaming Lai 0 Yunfei Yuan 0 Zhiliang Gao 0 Mengfeng Li 0 Jun Li 0 Libing Song 0 Xin Wei Wang, National Cancer Institute, United States of America 0 1 Department of Infectious Diseases, the Third Affiliated Hospital of Sun Yat-sen University , Guangzhou, Guangdong Province , China , 2 State Key Laboratory of Oncology in Southern China, Department of Experimental Research, Cancer Center, Sun Yat-sen University , Guangzhou, Guangdong Province , China , 3 Department of Microbiology, Zhongshan School of Medicine, Sun Yat-sen University , Guangzhou, Guangdong Province , China , 4 Department of Pathology, Cancer Center, Sun Yat-sen University , Guangzhou, Guangdong Province , China , 5 Department of Hepatobiliary Surgery, the First Affiliated Hospital of Sun Yat-sen University , Guangzhou, Guangdong Province , China , 6 Department of Hepatobiliary Surgery, Cancer Center, Sun Yat-sen University , Guangzhou, Guangdong Province , China , 7 Key Laboratory of Tropical Disease Control, Ministry of Education, Sun Yat-sen University , Guangzhou, Guangdong Province , China , 8 Department of Biochemistry, Zhongshan School of Medicine, Sun Yat- sen University , Guangzhou, Guangdong Province , China Background: The metastasis-associated in colon cancer 1 gene (MACC1) has been found to be associated with cancer development and progression. The aim of this study was to investigate the prognostic value of MACC1 in early-stage and AFP-normal hepatocellular carcinoma (HCC). Methods: mRNA and protein levels of MACC1 expression in one normal liver epithelial cells THLE3 and 15 HCC cell lines were examined using reverse transcription-PCR and Western blot. MACC1 expression was also comparatively studied in 6 paired HCC lesions and the adjacent non-cancerous tissue samples. Immunohistochemistry was employed to analyze MACC1 expression in 308 clinicopathologically characterized HCC cases. Statistical analyses were applied to derive association between MACC1 expression scores and clinical staging as well as patient survival. Results: Levels of MACC1 mRNA and protein were higher in HCC cell lines and HCC lesions than in normal liver epithelial cells and the paired adjacent noncancerous tissues. Significant difference in MACC1 expression was found in patients of different TNM stages (P,0.001). Overall survival analysis showed that high MACC1 expression level correlated with lower survival rate (P = 0.001). Importantly, an inverse correlation between MACC1 level and patient survival remained significant in subjects with early-stage HCC or with normal serum AFP level. Conclusions: MACC1 protein may represent a promising biomarker for predicting the prognosis of HCC, including in earlystage and AFP-normal patients. - Funding: This work was supported by the National Program on Key Basic Research Project (973 Program, No.2012CB519003); The National Science and Technique Major Project (201005022-2, 2012ZX09102101-017, 2012ZX10004213, 311030); The Key Science and Technique Research Project of Guangdong Province (2010B030600003, 2009010058); Natural Science Foundation of China (No. 81071780, 81272198, U1201121, 30900569, 81101864); The Science and Technology Department of Guangdong Province (No. S2011020002757); and Ministry of Education of China (20100171110080). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. Primary hepatocellular carcinoma (HCC) is the fifth most commonly diagnosed cancer and the third most common cause of cancer mortality worldwide [1]. HCC carcinogenesis involves aberrant changes in multiple molecular pathways and genetic as well as epigenetic factors, which consequently results in malignant transformation and progression of HCC [2,3]. Unfortunately, the long-term prognosis of patients with HCC remains unsatisfactory in spite of recent advances in surgical techniques and medical management. No specific signature of liver cancer gene expression has been reported to allow for patient-tailored therapy strategies. Hence, it is of great clinical value to further identify effective early markers for the diagnosis and prognosis of the disease as well as novel therapeutic targets. Clinically, prediction of prognosis plays an essential role in the assessment of HCC patients and optimal therapeutic options. Currently, both tumor- (e.g., alpha fetal protein (AFP), tumor size and portal vein thrombosis) and cirrhosis-based (mainly, the ChildPugh class) parameters are commonly employed to predict patient survival [4]. While tremendous effort has been made in identifying effective indicators for the prognostic prediction of HCC, the availability of clinically applicable biomarkers remains limited. Prediction of clinical prognosis of HCC is still reliant on conventional pathologic variables such as tumor size, tumor grade, lymph node, and distal metastasis status [5]. Furthermore, as the No. patients (%) = 1 Vital status (at follow-up) Death due to liver cancer cause Expression of MACC1 number of HCC patients who are in early stages of the disease, bear small tumor nodules (,3 cm), or display normal serum level of AFP is increasing, finding new prognosis prediction biomarker for these patients represents a major challenge in the clinic. MACC1, a novel regulator of tumor growth and metastasis, has recently been indicated as a potential prognostic factor for metastatic disease in HCC [6,7], lung adenocarcinoma [8], gastric carcinoma [9] and colorectal cancer [10,11]. Expression of MACC1 was found to be significantly upregulated in malignant tissues compared to normal tissues or adenomas, and induction of MACC1 might occur at the crucial step of transition from the benign to the malignant phenotype and can be allocated to the adenomacarcinoma sequence for colon cancer [10]. It is of note that hepatocyte growth factor (HGF) receptor, MET, is a transcriptional target of MACC1. MACC1 enhances proliferation and invasion as well as HGF-induced scattering of colon cancer cells in vitro and promotes tumor growth and metastasis of xenografted tumors in mouse [12]. Transduction of MACC1 mutants with the SH3 domain or the proline-rich motif deleted in colon cancer cells abrogated the above function of MACC1 [13]. It has been reported that MACC1 mRNA expression in HCC tissue was significantly higher than that in nonmalignant tissues, and high MACC1 mRNA expression correlated with more aggressive behavior in terms of shorter overall survival (OS) and disease-free survival [7]. However, the protein expression level of MACC1 in HCC was not investigated in these studies. In our current report, immunohistochemical analysis was performed to investigate the potential progno (...truncated)