Abstract:

Autophagy is a highly conserved catabolic process that delivers cytoplasmic constituents to lysosomes for degradation. Despite its importance, the mechanisms underlying matrine-induced autophagy in hepatoma cells remain largely elusive. This study aimed to elucidate the role of the p53/AMPK signaling pathway in mediating matrine-induced autophagy in human hepatocellular carcinoma cells. Initially, inhibition of p53 using the chemical inactivator PFT-α or RNA interference (RNAi) in combination with matrine treatment led to a pronounced increase in LC3Ⅱ levels, as evidenced by immunofluorescence and immunoblot analyses. Concomitantly, p53 silencing markedly enhanced AMPK phosphorylation, whereas expression of ULK1 and WIPI-1 showed a modest decline. In contrast, pharmacological modulation of AMPK using specific agonists and antagonists positively regulated ULK1 and WIPI-1 expressions in matrine-treated cells. Notably, WIPI-1 knockdown attenuated matrine-induced apoptosis and facilitated cell proliferation. Furthermore, comprehensive cDNA array profiling explored the molecular mechanisms of matrine in vivo, revealing significant regulation of SRGN and BCL11A, which was subsequently validated by quantitative real-time PCR (qPCR). Collectively, these findings uncovered intricate mechanisms underlying matrine-induced autophagy, highlighting the pivotal cross-talk between p53 and AMPK and identifying key downstream effectors, including ULK1 and WIPI-1. Moreover, the identification of SRGN and BCL11A as matrine-responsive genes suggested additional pathways through which matrine exerted its anti-tumor activity. By concurrently modulating autophagy-related signaling and specific oncogenic targets, matrine emerged as a promising multi-target therapeutic agent for hepatocellular carcinoma, warranting further exploration of its clinical potential.

Key words: Matrine, AMPK, ULK1, WIPI-1, Autophagy