海南地不容氧化克班宁调控PDK1/AKT通路诱导HepG-2细胞自噬机制OA

Based on previous findings that oxocrebanine from Stephania hainanensis H.S.Lo et Y.Tsoong(a Li ethnic medicine)induces autophagy in human hepatocellular carcinoma HepG-2 cells,this study further investigated the molecular mechanism,with a focus on verifying the regulatory role of phosphoinositide-dependent kinase-1(PDK1)-protein kinase B(Akt)signaling pathway in oxocrebanine-induced autophagy.The PDK1 gene was overexpressed in HepG-2 cells using liposome transfection to reversely validate the autophagic mechanism induced by oxocrebanine.Following treatment with oxocrebanine,the protein expression levels of the autophagy marker LC3-Ⅱ/LC3-Ⅰ ratio,total AKT,and phosphorylated AKT at Ser473(p-AKT)were detected by Western blotting.The results showed that oxocrebanine significantly increased the LC3-Ⅱ/LC3-Ⅰ ratio as well as the protein expression levels of AKT and p-AKT in PDK1-overexpressing cells,with extremely significant differences compared to the control group(P<0.01).In conclusion,oxocrebanine may activate the PDK1 signaling axis,promote AKT phosphorylation,and subsequently accelerate the conversion of LC3-Ⅰ to LC3-Ⅱ,ultimately inducing autophagy in HepG-2 cells.These findings provide new experimental evidence for the anti-hepatocellular carcinoma activity of active components derived from the Li ethnic medicine Stephania hainanensis.