The non-structural protein 5A (NS5A), which is unique among hepatitis C virus (HCV) non-structural proteins, is a proline-rich hydrophilic phosphoprotein that is essential for viral RNA replication and play a role in disrupting host intracellular signaling pathways. Although no intrinsic enzymatic activity has been ascribed to NS5A, this protein likely exerts its functions through interaction with viral and host cellular factors, as numerous NS5A-interacting cellular proteins have been identified. Recent studies using an HCV-infected cell culture system to model HCV RNA replication have suggested   that NS5A inhibition is a promising therapeutic strategy for the treatment of HCV. Most selected inhibitors against HCV replication derived from genotype 1b were targeted to NS5A. This review summarizes the progress made towards under-standing the role of NS5A in viral replication and HCV-mediated hepatocarcinoma for the development of possible inhibitors for HCV therapy.