Acetylation of MOB1 mediates polyphyllin II-reduced lysosome biogenesis in breast cancer by promoting the cytoplasmic retention of the YAP/TFEB coactivator complex

Background: Autophagy-lysosome abnormalities are associated with the malignant progression of cancer. Transcription factor EB (TFEB) is the master transcriptional regulator of the autophagy-lysosome machinery, and its abnormal activity is associated with autophagy-lysosome dysfunction. Polyphyllin II (PPII), an active steroidal saponin isolated from the rhizomes of Paris polyphylla, has been demonstrated to have antitumor activity. Purpose: Here, we explored the antitumor activity of PPII in breast cancer (BC) and further clarified its mechanism. Methods: Autophagosome was detected by transmission electron microscopy, an autophagy indicator system, and western blot. The effect of PPII on lysosomal activity was evaluated by flow cytometry, a lysosomal cathepsin activity assay, and acridine orange staining. The effect of PPII on the signaling pathway was evaluated by Western blot, gene expression measurement, gene alterations. The binding of PPII and MOB1 was examined through a drug affinity responsive target stability assay. The pharmacokinetic parameters of PPII were evaluated in Sprague-Dawley rats. Results: PPII exhibits therapeutic potential in BC by inducing the accumulation of autophagosome. PPII promotes the cytoplasmic retention of YAP/TFEB, which is responsible for the accumulation of autophagosome in BC. PPII activates Hippo signaling to promote cytoplasmic retention of YAP. PPII activates Hippo signaling by accelerating acetylation of MOB1 through a direct binding interaction. Conclusion: Taken together, these results confirm that acetylation of MOB1 mediates PPII-induced autophagosome accumulation in BC by promoting cytoplasmic retention of the YAP/TFEB coactivator complex. PPII is expected to be a drug candidate for the treatment of BC based on lysosomal biosynthesis.