Elevation of Plasminogen Activator Inhibitor-1 Promotes Differentiation of Cancer Stem-Like Cell State by Hepatitis C Virus Infection

Abstract

Plasminogen activator inhibitor-1 (PAI-1) is a critical factor that regulates protein synthesis and degradation. The increased PAI-1 levels are detectable in the serum of patients with chronic hepatitis C virus (HCV) liver disease. The differentiation state and motility of HCV-induced cancer stem-like cells (CSC) play a major role in severe liver disease progression. However, the role of PAI-1 in the pathological process of chronic liver diseases remains unknown. In this study, we determined how PAI-1 affects the differentiation of CSC state in hepatocytes upon HCV infection. We found that HCV infection induced the expression of PAI-1 while decreasing miR-30c expression in Huh7.5.1 cells. Similar results were obtained from isolated hepatocytes from humanized liver mice after HCV infection. Moreover, decreased miR-30c expression in HCV-infected hepatocytes was associated with the increased levels of PAI-1 mRNA and protein. Notably, the increased PAI-1 levels resulted in the activation of Protein Kinase B/AKT, a major mediator of cell proliferation, in HCV-infected hepatocytes along with the increased expression of CSC markers such as Human Differentiated Protein (CD) 133, Epithelial cell adhesion molecule (EpCAM), Octamer 4 (Oct4), Nanog, Cyclin D1, and MYC. Moreover, blockade of PAI-1 activity by miR-30c mimic and anti-PAI-1 mAb abrogated the AKT activation with decreased expression of CSC markers. Our findings suggest that HCV infection induces the CSC state via PAI-1-mediated AKT activation in hepatocytes. It implicates that the manipulation of PAI-1 activity could provide potential therapeutics to prevent the development of HCV-associated chronic liver diseases. IMPORTANCE The progression of chronic liver disease by HCV infection is considered a major risk factor for hepatocellular carcinoma (HCC), one of the major causes of death from cancer. Recent studies have demonstrated that increased CSC properties in HCV-infected hepatocytes are associated with the progression of HCC. Since proteins and miRNAs production by HCV-infected hepatocytes can play various roles in physiological processes, investigating these factors can potentially lead to new therapeutic targets. However, the mechanism of HCV associated progression of hepatocytes to CSC remains unclear. Here we identify the roles of PAI-1 and miR-30c in the progression of CSC during HCV infection in hepatocytes. Our data shows that increased secretion of PAI-1 following HCV infection promotes this CSC state and activation of AKT. We report that the inhibition of PAI-1 by miR-30c mimic reduces HCV associated CSC properties in hepatocytes. Taken together, targeting this interaction of secreted PAI-1 and miR-30c in HCV-infected hepatocytes may provide a potential therapeutic intervention against the progression to chronic liver diseases and HCC.

Epidemiological Characteristics of Primary Liver Cancer in Mainland China From 2003 to 2020: A Representative Multicenter Study

“…HCV itself might not be directly oncogenic. Chronic HCV infection causes hepatic inflammation, necrosis, metabolic disorders, steatosis, regeneration, and cirrhosis, thus facilitating the development of HCC by creating an immunosuppressive tumorigenic environment and activating cancer stem-like cells by proinflammatory factors like plasminogen activator inhibitor-1 ( 40 , 41 ). Other non-B and non-C risk factors including diabetes and NAFLD might also facilitate the development of HCC, mostly in elderly patients, and the overall survival rate is significantly better than that of HBV-HCC ( 42 ), possibly by inducing systemic and hepatic inflammation.…”

rs62139665 Polymorphism in the Promoter Region of EpCAM Is Associated With Hepatitis C Virus-Related Hepatocellular Carcinoma Risk in Egyptians

“…The higher expression of EpCAM observed in the HCV group compared to the control could be linked to the reported HCV-induced elevation of plasminogen activator inhibitor-1 (PAI-1). Increased expression of PAI-1 subsequent to HCV infection promoted the cancer stem-like cells (CSC) state in HCV-infected hepatocytes through the activation of the chief mediator of cell proliferation, protein kinase B, in consort with the increased expression of the CSC marker, EpCAM ( 31 ).…”

The role of oncolytic virotherapy and viral oncogenes in the cancer stem cells: a review of virus in cancer stem cells

“…In addition, plasminogen activator inhibitor-1 (PAI-1)-mediated AKT activation in hepatocytes caused by HCV infection promotes the CSC state. As a result, altering PAI-1 activity may provide new therapies to stop the onset of HCV-related chronic liver disorders [ 62 ]. In another study, HCV-infected hepatocytes show sphere formation on very low-adhesion plates with typical activation of the CSC signaling pathway in NOD-SCID IL2R gamma null (NSG) mice.…”