Study On The Effects Of Cardamonin Mediated By DAP1Regulating The Autophagy Of Ovarian Cancer Cells

Aim1To investigate the influence in proliferation and autophagy of ovarian cancer cells by DAP1-mediated cardamonin;2To study the mechanisms in treating ovarian cancer of cardamonin, which can provide theoretical and experimental basis for developing new mTOR targeted anticancer drugs.Methods1Cell cultureHuman ovarian cancer SKOV3cell lines and DAP1siRNA transfected SKOV3cells were cultured in McCoy’s5A medium with10%FBS,100U·mL-1penicillin and100μg·mL-1streptomycin at37℃with5%CO2. Cells were passaged when the adherent ones grew to70%-80%.2Groups and drugs interventionControl group, CAR group (10,20μmol·L-1), and RAP group (0.1μmol·L-1) were set in the two kinds of cells.3Measurements and methodology3.1Cells’ morphology and growth were observed by the inverted microscope;3.2Cells’ proliferation was measured by MTT method (λ=490nm);3.3The autophagy level of cells was detected by MDC staining method;3.4Examination of protein expression of DAP1and LC3were performed by Western blotting method.Results1siRNA of DAP1was transfected into SKOV3cells successfully. After transfection, both mRNA and protein expression of DAP1were decreased (P <0.01); 2DAP1siRNA transfected cells grew slowly compared with SKOV3cells;3Cells’ proliferation was significantly inhibited by CAR on SKOV3cells and DAP1siRNA transfected SKOV3cells (P <0.01);4The fluorescent spots with MDC staining and the expression of LC3Ⅱ protein significantly increased in DAP1siRNA transfected SKOV3cells compared to SKOV3cells (P <0.01);5After treated by CAR, the fluorescent spots with MDC staining and the expression of LC3Ⅱ protein significantly elevated in the two kinds of cells (P <0.01).ConclusionsInterference on DAP1can promote the occurrence of cells autophagy, which is positively correlated with the concentration of CAR.