Effect Of Celastrol On Proliferation And Pyroptosis Of Ovarian Cancer Cells And Its Mechanism

ObjectiveTo investigate the effect of celastrol on the proliferation and pyroptosis of human ovarian cancer cell line heya8 and its mechanism.MethodsMTT assay was used to determine the effects of Celastrol on HEYa8 cells at different concentrations(0,0.05,0.1,0.2,0.4,0.6,1.2 and 2.4 mg/L)and at different times(12h,24 h,36h,48 h and 72h),and the optimal concentration and time were calculated and analyzed.The experimental group(0.4 mg/L for 24hours)and the control group(untreated).Transwell method was used to detect the growth and migration of two groups of cells,colchicine treatment was used to detect the cell index of two groups of cells,cell counting instrument was used to detect the number of two groups of cells;Western blot method was used to detect protein: GSDMS-N,Caspase-1,NLRP3,ASC,IL-1beta,IL-18,PI3 K and AKT,and immunofluorescence double-labeling method was used to detect protein.The ratio of CD44/GSDMS-N double positive cells was detected.Finally,quantitative real-time polymerase chain reaction(qrt-pcr)was used to detect the m RNA levels of ASC and Caspase 1 of NLRP3 il-1 in HEYa8 cells.ResultsAt the concentration of 0.4mg/l and culture for 24 h,triptolide had the best effect;compared with the control group,the migration number,division index and cell count of heya8 cells in triptolide group were significantly reduced(P<0.05);the expression of apoptosis related proteins in triptolide group was increased(P<0.05);meanwhile,PI3 K / Akt signaling pathway was inhibited(P<0.05);IGF-1(PI3K specific agonist)was used to activate this pathway.They found that the level of charring decreased(P<0.05).ConclusionsTripterygium wilfordii can inhibit the growth of heya8 cells and induce cell scorching.The molecular mechanism may be related to the inhibition of PI3 K /Akt signaling pathway.