Osthole Inhibits Cervical Cancer Proliferation Through NQO1/ROS-mediated Cell Death

Objective: To explore the effect of osthole on cervical cancer He La cells,and clarify the mechanism of osthole inducing He La cell apoptosis,secondary necrosis and mitochondrial autophagy in vivo and in vitro.Lay theoretical foundation and scientific basis for the development of new cervical cancer treatment drugs.Methods: The effect of osthole on the proliferation of He La cells was detected by MTT method.The modes of cell death were detected by flow cytometry and Hoechst 33342/PI staining.The production of cellular ROS was detected by DCFH-DA staining,and mitochondrial membrane potential was detected by JC-1 staining.Differentially expressed proteins and related pathways were analyzed by label free proteomics.The expression of NQO1,apoptosis-,secondary necrosis-and mitophagy-related proteins was detected by western blotting.Finally,He La cells were used to establish a transplanted tumor model and set cisplatin as a positive control to test the inhibitory effect of osthole on tumors.Liver H&E staining was used to detect the toxic effects of osthole on the body.Western blotting was used to detect the expression of NQO1,apoptosis-,secondary necrosis-and mitophagy-related proteins in transplanted tumors.Results:(1)Osthole could significantly inhibit the proliferation of He La cells.Osthole induced apoptosis and up-regulates the expression of c-caspase 3 and c-PARP-1.(4)Osthole increased the expression of mitophagy-related factors PINK1,Parkin and LC3 Ⅱ/Ⅰ.(5)Osthole down-regulated the expression of NQO1 and induced the production of ROS.(6)NAC,DIC and Z-DEVD-FMK could reverse the apoptosis,secondary necrosis,mitochondrial autophagy and the expression of related proteins induced by osthole.(7)Osthole up-regulated the expression of apoptosis-related factors c-caspase 3 and c-PARP-1,secondary necrosis-related factor GSDME-N,and mitophagy-related factors PINK1,Parkin and LC3 Ⅱ/I.Conclusions: Osthole inhibited the proliferation of cervical cancer He La cells through NQO1/ROS-mediated apoptosis,secondary necrosis,and induced mitophagy...