Dihydroartemisinin Attenuates HBV-Induced Proliferation and Migration in LO2 cells via EMT Pathway Modulation

Abstract

Dihydroartemisinin (DHA), a derivative of artemisinin, possesses well-documented anti-inflammatory properties. This study investigated the inhibitory effects of DHA on hepatitis B virus (HBV)-induced cellular alterations in LO2 cells and elucidated underlying molecular mechanisms. Serum interleukin-6 (IL-6) levels were measured in HBV patients and analyzed for correlations with HBsAg, anti-HBs, and HBeAg. In vitro, LO2 cells were infected with clinical HBV serum samples (1 PEIU/ml, 72 hours) and treated with DHA (10 μM). Cell phenotypes were assessed using viability (MTT assay), proliferation (colony formation assay), and migration (scratch wound and Matrigel invasion assays). Protein expression related to proliferation (IL-6, tumor necrosis factor-α [TNF-α], programmed death ligand-1 [PD-L1]), apoptosis (Bcl-2 [B-cell lymphoma-2], Bax [Bcl-2-associated X protein]), cell cycle (p21 [cyclin-dependent kinase inhibitor 1A], cyclin D1), and epithelial-mesenchymal transition (E-cadherin, α-catenin, N-cadherin) were quantified by Western blotting. Serum IL-6 positively correlated with HBsAg/HBeAg and negatively correlated with anti-HBs. HBV infection increased cell viability, proliferation, and migration capacity, and upregulated IL-6, TNF-α, PD-L1, Bcl-2, cyclin D1, and N-cadherin protein expression, while downregulating Bax, p21, E-cadherin, and α-catenin. DHA treatment suppressed HBV-induced malignant phenotypes and reversed these protein expression changes. These findings demonstrate that HBV activates an IL-6/TNF-α/PD-L1 signaling axis to drive malignant transformation, while DHA attenuates this process through multi-target inhibition, supporting its potential therapeutic application in HBV-associated liver disease.