Investigation On The Antiproliferation Of Cardamonin Through Inhibiting The Protein Expression Of GLS Via Raptor On Ovarian Cancer Cells

AimsTo clarify the effects and mechanism of cardamonin（CAR）on antiproliferationand apoptosis induction through decreasing the protein expression of glutaminase（GLS）via Raptor on ovarian cancer cells.Methods1 Cell cultureCells including SKOV3,Raptor knockout SKOV3（constructed through the CRISPR-Cas9 technology）and A2780 cells were cultured in Mc Coy’s 5A and RPMI-1640 medium.All medium were supplemented with 10%fetal bovine serum（FBS）,100 mg·m L^-1of streptomycin and 100 U·m L^-1of penicillin.Cells were cultured at 37℃with 5%CO₂.2 Detection indexes and methodology2.1 Cell viability was measured by CCK8;2.2 Proteins expression were detected by Western blotting;2.3 Cell apoptosis and Reactive oxygen species（ROS）level were detected by flow cytometry.Results1 Cells proliferation of SKOV3 and A2780 were inhibited by CAR in a concentration-dependent manner（P<0.05）.CAR induced apoptosis of SKOV3 cells and increased the expression of Cleaved caspase-3 and-9 in a concentration-dependent manner（P<0.05）.CAR induced ROS production in ovarian cancer cells（P<0.05）.NAC reversed the inhibitory effect of CAR on proliferation of ovarian cancer cells（P<0.05）.2 The proliferation of SKOV3 and A2780 cells were glutamine dependent.SKOV3 cells rely more on glutamine（P<0.05）.The proliferation of SKOV3 and A2780 cells induced by glutamine was inhibited when treated with CAR（P<0.05）.3 The protein expression of GLS was inhibited by CAR in a concentration-dependent manner in SKOV3 cells（P<0.05）.4 The protein expression of Raptor and phosphorylation of mTOR and S6K1were decreased by CAR in a concentration-dependent manner（P<0.05）.5 The protein expression of Raptor,p-S6K1 and GLS were decreased in Raptor knockout SKOV3 cells.Nevertheless,CAR had no obvious effects on the protein expression of p-S6K1 and GLS in Raptor knockout SKOV3 cells which were decreased in the normal SKOV3 cells（P<0.05）.ConclusionCAR inhibits proliferation,induces apoptosis and promotes ROS production in ovarian cancer cells.The underlying mechanism is related to the inhibitory effcet on Raptor expression by CAR,which leads to the decrease of GLS expression.