Erinacine Facilitates The Opening Of The Mitochondrial Permeability Transition Pore Through The Inhibition Of The PI3K/Akt/GSK-3? Signaling Pathway In Human Hepatocellular Carcinoma

Objectives Erinacine,which is extracted from the medicinal mushroom Hericium erinaceus,is known to play anticancer roles in human cancers.The following study aims to investigate that erinacine can induce the opening of the mitochondrial permeability transition pore(mitochondrial Permeability Transition Pore,m PTP)in hepatocellular carcinoma(Human Hepatocellular Carcinoma,HCC)Hep G-2 cells through inhibiting the PI3K/Akt/GSK-3? signaling pathway,leading to apoptosis of Hep G-2 cells,and highlights the applicability of erinacine in HCC treatments.Methods In this experiment,Hep G2 cells originated from Human Hepatocellular Carcinoma tissues and purchased from the American Type Culture Collection(ATCC)were selected as the research object.First of all,in order to explore whether the different concentrations of erinacine(100?M,300?M,500?M)and 1?M LY294002(PI3K/Akt/ GSK-3? signaling pathway inhibitor)and the combination of 500?M erinacine +1?M LY294002 can inhibit PI3 K / Akt / GSK-3? signaling pathway and the degree of inhibition,RT-q PCR and Western blot techniques were used to detect the relative m RNA and protein expression levels and phosphorylation levels of PI3 K,Akt,GSK-3?.Then,we discussed whether erinacine can induce the opening of mitochondrial Permeability Transition Pore(m PTP).The above five groups and the 500?M erinacine + Cs A(the inhibitor of m PTP opening)group were used JC-1 for fluorescent staining and the ratio of red/green fluorescence intensity is measured by using laser scanning confocal microscopy to judge the change of mitochondrial membrane potential(??m),and judge the degree of the opening of m PTP.Finally,in order to explore whether erinacine can inhibit the growth of Hep G-2 cells and the degree of inhibition of the above five experimental groups,MTT,wound healing assay,and flow cytometry,were used to detect cell viability,migration,cycle,and apoptosis respectively.Results 1 Different concentrations of erinacine or LY294002 reduced the m RNA and protein expression levels and phosphorylation levels of PI3 K,Akt,and GSK-3?(P<0.05),suggesting that erinacine can inactivate the PI3K/Akt/GSK-3? signaling pathway.2 Different concentrations of erinacine or LY294002 can reduce the red/green fluorescence intensity ratio of Hep G2 cells.Compared with 500?M erinacine,the red/green fluorescence intensity ratio of the 500?M erinacine + Cs A(the inhibitor of m PTP opening)group was increased significantly(P<0.05),indicating that Hericin can reduce the mitochondrial membrane potential,suggesting that erinacine can induce m PTP opening by inactivated the PI3K/Akt/GSK-3? signaling pathway.3 Erinacine or LY294002 increased the inhibition rate of Hep G2 cells(P<0.05),suggesting that erinacine can inhibit cell proliferation;erinacine or LY294002 reduced the migration rate of Hep G2 cells(P<0.05),suggesting that erinacine can inhibit cell migration;erinacine or LY294002 increased the number of Hep G2 cells in G1 phase and decreased the number of cells in S phase,and the apoptosis rate of Hep G2 cells wree increased(P<0.05),suggesting that erinacine can inhibit the cell cycle and promote apoptosis.4 Erinacine works in a concentrationdependent manner,the higher the concentration,the stronger the effect,and erinacine can enhance the effect of LY294002.Conclusions The results from the study demonstrated that erinacine can induce m PTP opening by inactivating the PI3K/Akt/GSK-3? signaling pathway,inhibit cell proliferation,migration,and invasion,promote cell apoptosis,and prevent the progression of HCC.Figure13;Table2;Reference 167...