Study on the mechanism of cinobufotalin in inducing ferroptosis of hepatoma cells by inhibiting the expression
Objective To investigate the mechanism of cinobufotalin in inducing ferroptosis of hepatoma cells by inhib-iting the expression of glutathione synthetase(GSS).Methods Hepatoma cells were treated with different concentrations of cinobufotalin(0,2,4,6,8 μg·mL-1)or in combination with deferoxamine(DFO)50 μmol·L-1,glutathione(GSH)5 mmol·L-1 for 48 h,respectively.CCK-8 was used to detect cell viability.Reactive oxygen species(ROS),malondialdehyde(MDA)and GSH assay kit were used to detect ROS,MDA,and GSH levels in tumor cells after treatment of cinobufotalin(8 µg·mL-1).Quantitative real-time PCR(QRT-PCR)and Western blotting were used to detect the effect of cinobufotalin on GSS expression in hepatoma cells.RNA interference technology was used to explore the effect of GSS on GSH level and proliferation potential of hepatoma cells.Results Compared with cinobufotalin treatment alone,the DFO could partially reverse the inhibitory effect of cinobufotalin on hepatoma cells proliferation.Compared with the control group,the ROS and MDA levels were increased and the GSH level was decreased in cinobufotalin treatment group.At the same time,the study showed that GSH could partially reverse the inhibitory effect of cinobufotalin on the proliferation of hepatoma cells.Further mechanism study showed that cinobufotalin could inhibit the expression of GSS protein.The results of RNA in-terference experiment showed that depletion of GSS could reduce GSH level,knockdown of GSS could impair the reverse effect of DFO on the proliferation inhibition of cinobufotalin,and knockdown of GSS could inhibit the growth of hepatoma cells,and enhance the blocking effects of cinobufotalin on cell proliferation of liver cancer cells.Conclusion Cinobufotalin can induce ferroptosis in hepatoma cells,and the mechanism is related to GSS intervention.This study provides a theoretical basis for the clinical application of cinobufotalin,and plays an active role in promoting the development of anti-tumor drugs in traditional Chinese medicine.
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维普
