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Archive > Volume 31 Issue 5 > 2024,31(5):445-451. DOI:10.3872/j.issn.1007-385X.2024.05.003 Prev Next
Basic Research
Ginsenoside Rb1 promotes oxeiptosis in hepatocellular carcinoma HepG2 cells through KEAP1/PGAM5/AIFM1 pathway
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Abstract:
Objective: To investigate the effect of ginsenoside Rb1 (Gn-Rb1) on oxeiptosis in hepatocellular carcinoma (HCC) HepG2 cells and its possible molecular mechanism. Methods: Bioinformatic was used to analyse the effect of the expression of PGAM5, a key gene of oxeiptosis, on the survival time of HCC patients. HCC tissues and paracancerous tissues of 8 HCC patients admitted into Liaoning Provincial Tumor Hospital were selected, and the expressions of oxeiptosis-related gene proteins and mRNAs were detected by WB assay and qPCR. HepG2 cells were randomly divided into the control group and the Gn-Rb1 group (intervened with 200 μmol/L Gn-Rb1). The effects of Gn-Rb1 on the colony formation ability and the migration ability of HepG2 cells were detected by the cell clone formation assay and the cell scratch assay, respectively. ELISA was used to detect the effect on the level of ROS. Microplate was used to detect the effect on the level of LDH; the effects of Gn-Rb1 on the expressions of key gene proteins and mRNAs were detected by WB assay and qPCR. Results: Bioinformatic analysis revealed that, compared with low-expression patients, patients with PGAM5 high-expression hepatocellular carcinoma had longer overall survival time (P<0.05). In clinical hepatocellular carcinoma and paracancerous tissue samples, it was found that compared with paracancerous tissues, at protein and mRNA levels, the expressions of KEAP1 and PGAM5 in tumor tissues were significantly lower; the expression of NRF2 was significantly higher (all P<0.01); and the expression of p-AIFM1 was significantly higher (P<0.05). After HepG2 cells were intervened with 200 μmol/L Gn-Rb1, compared with those in the control group, the migratory and colony-forming abilities of HepG2 cells in the Gn-Rb1 group reduced significantly (all P<0.01), and the level of LDH increased significantly (P<0.05). Compared with the control group, at mRNA and protein levels, the expressions of KEAP1 and PGAM5 in the Gn-Rb1 group were significantly higher (all P<0.05); the expression of NRF2 was significantly lower (all P<0.05); and the expression of p-AIFM1 protein was significantly lower (P<0.01). Conclusion: Oxeiptosis is inhibited in hepatocellular carcinoma tissues, and Gn-Rb1 promotes oxeiptosis in HCC HepG2 cells by regulating the KEAP1/PGAM5/AIFM1 pathway, thus suppressing the proliferation and migration abilities of HepG2 cells.
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