Investigation Of The Mechanism Of Cardamonin On Antiproliferation And Migration Induced By Oxidative Stress On Ovarian Cancer Cells

Aim Investigating the role and mechanism of cardamonin(CAR)inhibiting the proliferation and migration of ovarian cancer cells by reducing the expression of Raptor protein inducing oxidative stress.Method1 Cell culture: The human ovarian cancer SKOV3 cell line,the Raptor knockout SKOV3 cell line and the human ovarian cancer A2780 cell line were prepared with Mc Coy’s 5A and 1640 containing 10% fetal bovine serum(FBS)and 1% penicillin-streptomycin,respectively.Culture medium,culture conditions were 37 ℃,5% CO2 constant temperature incubator.2 Indicators and Methods2.1 The levels of intracellular ROS was detected by Flow cytometry;2.2 The cell viability was evaluated by CCK8 assay;2.3 Wounding healing assay was applied to observe cell migration ability;2.4 The intracellular protein was quantified by Mass spectrometry;2.5 Western blotting was performed to detect the expression of S6K1,p-S6K1,mTOR,p-mTOR,MAPK(ERK1/2),p-ERK1/2,Raptor,β-Actin protein in cells.Results1.CAR induces the production of ROS and inhibit the proliferation and migration of ovarian cancer SKOV3 and A2780 cells;the active oxygen scavenger NAC can reduce the inhibitory effect of CAR on the proliferation and migration of ovarian cancer cells;2.CAR has a significant impact on the MAPK signaling pathway and inhibits the expression of p-ERK1/2,but does not affect its total protein expression;3.CAR inhibits the expression of p-mTOR and its downstream p-S6K1.The mTOR agonist MHY1485 pretreatment can antagonize the phosphorylation inhibitory effect of CAR on mTOR and its downstream substrate S6K1;4.Raptor knockout SKOV3 cells have significantly lower Raptor protein expression;compared with wild-type cells,Raptor knockout cells have increased ROS production,and cell viability and migration ability are significantly reduced;5.CAR can significantly reduce the protein expression of p-mTOR,Raptor,p-S6K1 and p-ERK1/2;after Raptor was knocked out,the inhibitory effect of CAR on p-ERK1/2,p-mTOR,and p-S6K1 is weakened.Conclusions1 CAR induces oxidative stress and inhibits the proliferation and migration in ovarian cancer cells;2 Raptor mediates CAR’s inhibition of ERK1/2 and mTORC1 to induce cell oxidative stress.