Original ArticlesHepatitis C virus NS3 protein enhances cancer cell invasion by activating matrix metalloproteinase-9 and cyclooxygenase-2 through ERK/p38/NF-κB signal cascade
Introduction
Hepatitis C virus (HCV) infection is a global health problem that causes acute and chronic hepatitis, liver fibrosis, cirrhosis, and hepatocellular carcinoma (HCC) [1], [2]. The viral genome is a positive-strand RNA containing an open reading frame encoding for a precursor polyprotein that is cleaved by proteases into structural and nonstructural proteins [3], [4]. HCV replication is regulated by cellular factors, signal cascades, and viral proteins [5], [6], [7], [8], [9]. The non-structural protein 3 (NS3) of HCV possesses serine protease activity essential for viral protein processing and helicase activity required for virus replication [10], [11]. NS3 also plays a role in regulating cell proliferation, implicating its involvement in carcinogenesis [12], [13].
Cancer metastasis involves complex events, including proteolytic degradation of the components of extracellular matrix (ECM) [14], [15]. The well known ECM-degrading enzymes are matrix metalloproteinases (MMPs), a family of endoproteinases contributing to tumor invasion, metastasis, and angiogenesis [16], [17], [18], [19], [20]. Among them, gelatinase-B (MMP-9) is the key enzymes degrading type IV collagen, a major component of the basement membrane, and is implicated in macrophage differentiation, inflammation, and tumor invasion [21], [22], [23], [24]. Cyclooxygenase-2 (COX-2) is an enzyme that catalyzes the conversion of arachidonic acid to prostaglandin H2 [25]. COX-2 is expressed normally in the brain and spinal cord where it determines the inflammatory response mediated by interleukin-1β [26], induced in various tissues in response to inflammatory and tumorigenic signals [27], [28], and during viral infection [29], [30], [31], [32]. COX-2 regulates the process of fever development and controls the apoptotic pathway to help tumor cells evading apoptosis [33], [34]. The intrahepatic level of COX-2 was enhanced by HCV core and NS5A proteins, indicating an association with progressive hepatic fibrosis [35], and COX-2 expression and PGE2 production were regulated by HCV NS3, suggesting a role in inflammatory response [36].
Here, we initially showed that MMP-9 was activated in the sera of HCV-infected patients and hepatoma cells, and demonstrated that HCV NS3 protein activated MMP-9 and COX-2 expression. The mechanism by which NS3 activates MMP-9 expression was revealed, and the roles of NS3 in regulating hepatoma cell invasion and migration were determined. This study provides new insight into the roles of NS3, MMP-9 and COX-2 in regulating cancer cell invasion, and perhaps into the development of therapeutics against the viral infection.
Section snippets
Clinical samples
Blood samples were obtained from 14 patients with HCV infection who were admitted to Rennin Hospital of Wuhan University, China. All patients were confirmed HCV positive, but negative for other pathogens. A control group consisting of 14 healthy individuals with no history of liver disease was randomly selected from a Wuhan blood donation center, China. Written informed consent was obtained from each patient. The study was conducted according to the principles of the Declaration of Helsinki and
MMP-9 was up-regulated in HCV-infected patients and hepatoma cells
To investigate the correlation between HCV infection and MMP-9 expression, we evaluated MMP-9 level in HCV-infected patients and HCV-negative individuals. ELISA results indicated that MMP-9 protein were significantly higher (P < 0.05) in the sera of HCV-positive group (n = 14, mean ± 595.3 ng/ml) than HCV-negative group (n = 14, mean ± 200.1 ng/ml) (Fig. 1A). The effect of HCV infection on the expression of MMP-9 and COX-2 was also evaluated at the cellular level. Western blots showed that
Discussion
HCV infection is one of the leading causes of liver fibrosis and cirrhosis and is an increasingly important factor in the etiology of HCC [41]. Fibrotic liver disease is characterized by changes in tissue architecture and extracellular matrix composition that ultimately compromise organ function. Results from several recent studies suggested that epithelial–mesenchymal transition (EMT) may be an important mechanism for HCC metastasis [42], [43]. While, carcinogenesis is a complex process
Conflict of interest
None.
Acknowledgments
This work was supported by research grants from the Major State Basic Research Development Program (973 Program) (2012CB518900), the National Natural Science Foundation of China (31230005 and 81171525), the National Mega Project on Major Infectious Disease Prevention (2012ZX10002006-003 and 2012ZX10004-207), the National Mega Project on Major Drug Development (2015ZX09102-007) to JW; and the Natural Science Foundation Hubei Province of China (2012FFB04903), the Open Research Fund Program of the
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Lili Lu and Qi Zhang contributed equally to this work.