TNFAIP1 suppresses hepatocellular carcinoma progression via the PXR/CYP3A4 signaling axis

Hepatocellular carcinoma (HCC) is characterized by poor prognosis and limited effective treatment options, necessitating a deeper understanding of its pathogenesis. This study focuses on tumor necrosis factor-α-induced protein 1 (TNFAIP1) and cytochrome P450 3A4 (CYP3A4) in HCC, aiming to investigate their association and functional roles in tumor progression. Bioinformatics analyses and experimental validation revealed that both TNFAIP1 and CYP3A4 are downregulated in HCC, and that TNFAIP1 positively regulates CYP3A4 expression. TNFAIP1 knockout not only decreased CYP3A4 expression but also significantly impaired the ability of rifampicin (RIF), an upstream nuclear receptor-pregnane X receptor (PXR) agonist, to induce CYP3A4, indicating that TNFAIP1 is an essential regulator of PXR/CYP3A4 pathway. Co-immunoprecipitation (Co-IP) experiment further confirmed the direct interaction between TNFAIP1 and PXR. TNFAIP1 knockout promoted HCC cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT), while suppressing apoptosis; these effects were partially attenuated by pharmacological activation of PXR or genetic overexpression of CYP3A4. In vivo experiments demonstrated that overexpression of Tnfaip1 upregulated the Pxr/Cyp3a11 pathway and inhibited tumor growth, whereas Tnfaip1 knockout suppressed this pathway. This study identified TNFAIP1-PXR-CYP3A4 as a novel tumor-suppressive axis in HCC, providing potential molecular targets for HCC diagnosis and treatment.