MACC1 upregulation promotes gastric cancer tumor cell metastasis and predicts a poor prognosis

Xie et al. / J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2016 17(5):361-366 361 Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology) ISSN 1673-1581 (Print); ISSN 1862-1783 (Online) www.zju.edu.cn/jzus; www.springerlink.com E-mail: MACC1 upregulation promotes gastric cancer tumor cell metastasis and predicts a poor prognosis* Qiu-ping XIE§†1, Cheng XIANG§†1, Gang WANG2, Ke-feng LEI3, Yong WANG†‡1 (1Department of General Surgery, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China) (2Department of Colon Surgery, the Cancer Hospital of Zhejiang Province, Hangzhou 310022, China) (3Department of General Surgery, Zhejiang Provincial People’s Hospital, Hangzhou 310001, China) † E-mail: ; ; Received Sept. 27, 2015; Revision accepted Dec. 27, 2015; Crosschecked Apr. 15, 2016 Abstract: In various studies, metastasis associated with colon cancer 1 (MACC1) has been frequently reported to be abnormally highly expressed in human lung cancer, colon cancer, and hepatocellular carcinoma. Our study focuses on the association of MACC1 expression with gastric cancer (GC). During our experiment, the MACC1 expression was tested in 105 GC samples using an immunohistochemical (IHC) method. The clinical characteristics and prognosis of these patients were summarized. During analysis, MACC1 distribution in GC samples with distant metastasis was higher than that in normal samples and in tumors with no dissemination. Subsequently, a lower 5-year survival rate had a strong correlation with high MACC1 expression. As a consequence, the present results suggest that MACC1 is more frequently expressed in a poor prognosis phenotype of GC and acts as a promising prognostic prediction parameter for GC. Key words: Gastric cancer, MACC1, Prognosis, Metastasis http://dx.doi.org/10.1631/jzus.B1500236 CLC number: R735.2 1 Introduction Gastric cancer (GC) is the fourth most frequently diagnosed cancer and ranks as the third leading cause of death from cancer worldwide. In 2011, a global analysis showed that 989 600 new GC cases and as many as 738 000 patient deaths occurred in 2008. Among the population, 70% of cases occur in developing countries, especially in eastern Asia (Jemal et al., 2011). Over two-thirds of cases present with advanced or inoperable disease once diagnosed, and the prognosis is poor (MacDonald, 2006). Despite the ‡ Corresponding author The two authors contributed equally to this work * Project supported by the National Natural Science Foundation of China (No. 30901445) ORCID: Qiu-ping XIE, http://orcid.org/0000-0002-4658-5935; Yong WANG, http://orcid.org/0000-0002-2034-6709 © Zhejiang University and Springer-Verlag Berlin Heidelberg 2016 § wide usage of advanced surgical techniques (Lee et al., 2008) and chemotherapy (Sun et al., 2009), the overall recovery rate for patients continues to remain poor (Morabito et al., 2009). GC pathogenesis is a complex, multistage, and heritage-related process. So far, the researches associated with GC mechanisms are far from understood. Various pathological and epidemiological studies have provided evidence that genetic factors play a crucial role in gastric carcinogenesis (González et al., 2002; Hamajima et al., 2006). As a feature of malignant tumors, metastasis is mainly related to GC prognosis and the major factors of GC recurrence. Furthermore, lymph node and peritoneal metastasis are highly associated with a poor GC prognosis. We are very interested in the undiscovered molecular changes in distant metastasis tumors. In colon cancer, metastasis associated with colon cancer 1 (MACC1) gene was first identified to be differentially expressed by a gene expression contrast 362 Xie et al. / J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2016 17(5):361-366 search by comparing normal colon mucosa, primarystage tumors, and metastases lesions (Stein et al., 2009). It induces tumor proliferation, metastasis, and invasion in in vitro colon epithelium cancer cell culture, as well as in colon cancer patients (Arlt and Stein, 2009). Besides this, MACC1 acts as the main regulator factor of the HGF/Met pathway, and influences cell activity through downstream MAPK upregulation (Stein et al., 2009). In previous studies, high MACC1 expression was found to be present in various cancer tissues including colon cancer (Shirahata et al., 2010a), hepatocarcinoma (Shirahata et al., 2011), and lung adenocarcinoma (Chundong et al., 2011; Shimokawa et al., 2011). Accumulating evidence suggests that the high expression of MACC1 has been proved to be associated with increased metastasis risk and poorer patient survival in various types of cancers including bladder urothelial carcinoma (Xu et al., 2015), cervical cancer (Zhou et al., 2015), and colorectal cancer (Weidle et al., 2015). Recently, upregulation of MACC1 expression was reported in GC (Shirahata et al., 2010b). MACC1 was reported to play a key role in GC metastasis (Wang et al., 2013), epithelial-mesenchyal transition (EMT) phenomena (Huang et al., 2015), and angiogenesis (Sun et al., 2015). Until now, the role of MACC1 in GC has still remained controversial (Ge et al., 2011). During our work, we investigated MACC1 expression in surgical specimens from 105 GC patients, and we identified the correlation between MACC1 expression and various clinic pathological parameters. 2 Materials and methods 2.1 Patients and tissue samples GC tissue was obtained from 105 patients who were pathologically proven and underwent surgical operation between January 2004 and December 2006 in our hospital. None of the 105 patients had received neoadjuvant before operation. The tissue samples were formalin-fixed and paraffin-embedded following surgical removal, then were cut into thick sections (4 µm) and mounted onto glass slides for further protein expression analysis. The clinical characteristics of these patients, such as gender, age, tumor location, differentiation, and tumor-node-metastasis (TNM) stage, are summarized in Table 1. Follow-up by consultation of the case documents or by telephone was performed until death or June 2011, whichever came first. The clinical stage of the tumors was determined according to the TNM classification of the International Union Against Cancer (2009) (Edge and Compton, 2010). Thirty normal tissue specimens from gastritis patients without malignancy undergoing endoscopic biopsy were collected as blank controls. The present study was approved by the Ethical Committee of the Second Affiliated Hospital of Zhejiang University and the Zhejiang Provincial People’s Hospital, China. 2.2 Immunohistochemical staining of MACC1 Immunohistochemical (IHC) examination of MACC1 was performed using rabbit anti-MACC1 polyclonal antibody (Abcam, Hong Kong, China) on the specimens described previously. Tissue sections were first incubated at 60 °C for 2 h, followed by deparaffinizing them in xylene; then they were rehydrated in a list of graded alcoho (...truncated)