Activation of TGF-ß1 promoter by hepatitis C virus-induced AP-1 and Sp1: role of TGF-ß1 in hepatic stellate cell activation and invasion.

Our previous studies have shown the induction and maturation of transforming growth factor-beta 1 (TGF-ß1) in HCV-infected human hepatoma cells. In this study, we have investigated the molecular mechanism of TGF-ß1 gene expression in response to HCV infection. We demonstrate that HCV-induced transcription factors AP-1, Sp1, NF-κB and STAT-3 are involved in TGF-ß1 gene expression. Using chromatin immunoprecipitation (ChIP) assay, we further show that AP-1 and Sp1 interact with TGF-b1 promoter in vivo in HCV-infected cells. In addition, we demonstrate that HCV-induced TGF-ß1 gene expression is mediated by the activation of cellular kinases such as p38 MAPK, Src, JNK, and MEK1/2. Next, we determined the role of secreted bioactive TGF-ß1 in human hepatic stellate cells (HSCs) activation and invasion. Using siRNA approach, we show that HCV-induced bioactive TGF-ß1 is critical for the induction of alpha smooth muscle actin (α-SMA) and type 1 collagen, the markers of HSCs activation and proliferation. We further demonstrate the potential role of HCV-induced bioactive TGF-ß1 in HSCs invasion/cell migration using a transwell Boyden chamber. Our results also suggest the role of HCV-induced TGF-ß1 in HCV replication and release. Collectively, these observations provide insight into the mechanism of TGF-ß1 promoter activation, as well as HSCs activation and invasion, which likely manifests in liver fibrosis associated with HCV infection.