Study on the effects of Polyphyllin I and Curcumin on liver cancer based on the cross-action of ferroptosis and energy metabolism

Abstract

Background: To investigate the effect and mechanism of Polyphyllin I(PPI) and Curcumin(Cur) on human liver cancer HepG2 and HepG2.2.15 cells. Methods: Download the hepatocellular carcinoma specimens and normal control specimens from the TCGA database, take the intersection with ferroptosis-related genes, and screen the differentially expressed ferroptosis genes; again, make the intersection with the selected differential genes related to energy metabolism; conduct survival analysis; construct prognosis Risk scoring model and evaluation of model performance; through molecular docking to verify the binding effect of PPI, Cur and Ribonucleoside-diphosphate reductase subunit M2(RRM2), SRC(SRC), Acetyl-CoA carboxylase alpha(ACACA) and other genes. Human hepatocellular carcinoma cells were cultured in vitro, PPI and Cur intervened, and Cell Counting Kit-8(CCK-8) was used to detect cell inhibition rate; FeRhoNox-1 fluorescent probe staining to observe the intracellular Fe 2+ status; lactate dehydrogenase (LDH) release Experiment to detect cell LDH release rate; JC-1 staining to detect mitochondrial membrane potential; reactive oxygen species(ROS) kit to detect ROS level;Western blotting (WB) to detect RRM2 and SRC , ACACA and other genes protein expression levels. Results: Through screening, 25 differential genes related to ferroptosis and energy metabolism in liver cancer were obtained;; through survival analysis, three ferroptosis-related genes, such as RRM2, SRC, and ACACA, were obtained，the results showed that these three genes showed high expression and predicted poor overall survival(OS) and disease-free survival(DFS); molecular docking results showed that PPI, Cur It has good affinity with RRM2, SRC and ACACA. The results of in vitro experiments show that PPI and Cur inhibit cell proliferation in a concentration-dependent manner ( P <0.01). PPI and Cur can significantly increase the intracellular Fe2+ concentration, LDH release rate and intracellular ROS levels of HepG2, HepG2.2.15 ( P <0.01), and the effect on mitochondrial membrane potential was significantly lower than that of the blank group ( P <0.01), and significantly down-regulated the protein expression levels of RRM2, SRC, and ACACA ( P <0.01). Conclusion: The high expression of RRM2, SRC, ACACA and other three genes related to ferroptosis and energy metabolism in liver cancer indicate poor OS and DFS; PPI and Cur can up-regulate the LDH release rate, ROS and Fe 2+ levels of liver cancer cells, and down-regulate the cell mitochondrial membrane potential and other methods to inhibit the proliferation of liver cancer cells; and down-regulate the expression of RRM2, SRC, ACACA and other proteins to affect the prognosis of liver cancer.