This study aimed to evaluate the anti-pancreatic cancer effects of flavonoids in Plumula Nelumbinis. High-performance liquid chromatography/quadrupole time-of-flight mass spectrometry showed that apiin, rhoifolin, and vitexin were three principal components in total flavonoids derived from Plumula Nelumbinis, with vitexin being the most abundant component. Cell viability assay revealed that apiin, rhoifolin, and vitexin could inhibit proliferation of PANC-1 and ASPC-1, with rhoifolin showing the maximum inhibitory effect. Rhoifolin inhibited cell proliferation and promoted apoptosis of pancreatic cancer cells, which was associated with up-regulated JNK and p-JNK as well as down-regulated p-AKT. Rhoifolin also inhibited cell migration and invasion, and increased the antioxidant capacity in PANC-1 and ASPC-1. Besides, AKT activator (SC79) or JNK inhibitor (SP600125) effectively reversed the anticancer effects of rhoifolin in pancreatic cancer. Quantitative proteomics analysis showed that rhoifolin altered proteomic profiles in pancreatic cancer cells. Western blot analysis showed that rhoifolin down-regulated transforming growth factor beta 2 (TGF-β2), the regulator of proteoglycan synthesis, with the concomitant down-regulation of phosphorylated SMAD family member 2 (SMAD2), the downstream effector of TGF-β2. In conclusion, rhoifolin regulates the AKT/JNK/caspase-3 and TGF-β2/SMAD2 signaling pathways, which may contribute to its anti-pancreatic cancer effects.

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