Inhibitory Effect Of LY294002 On Biological Behavior Of Ovarian Cancer Cells Through PI3K/AKT/mTOR Signal Pathway

Objective: This study observed the effect of LY294002(PI3K inhibitor)on the biological behavior of ovarian cancer cell SKOV3 by culturing cells in vitro,and explored the possible mechanism of PI3K/AKT/m TOR signaling pathway in ovarian cancer cells.Methods: Ovarian cancer cell line SKOV3 was cultured in vitro and divided into control group and LY294002 treatment group(concentrations of 5umol/L,10umol/L,20umol/L and 40umol/L,respectively).After treatment for 0h,24 h,48h and 72 h,The proliferation of SKOV3 cells was measured by CCK-8 cell proliferation assay,and the IC50 value was calculated to screen the appropriate concentration for subsequent experiments.The effect of LY294002 on the adhesion ability of SKOV3 cells was tested by cell adhesion experiment.Cell scratch assay and Transwell migration assay were used to detect the effect of SKOV3 cell migration ability in control group and LY294002 treatment group after 48 h treatment.Transwell invasion assay was used to compare the effects of the control group and LY294002 treatment group on the invasion ability of SKOV3 cells.The effects of LY294002 on the protein expression of PI3 K,p-PI3 K,AKT,p-AKT,m TOR and p-m TOR in SKOV3 cells were detected by Western blot assay.Results:1.The results of CCK-8 experiment showed that LY294002 could inhibit the proliferation of SKOV3 ovarian cancer cells,and the inhibitory effect was gradually enhanced with the increase of LY294002 concentration and the extension of the action time,in a time-and dose-dependent manner.The absorbance of LY294002 treatment group was lower than that of control group,and the differences were statistically significant(P<0.05).The inhibition rate of 5umol/L LY294002 treatment group was(31.03±12.99)% for 24 h,(33.81±8.2)% for 48 h and(42.29±4.65)% for 72 h.The inhibition rate of 10umol/L LY294002 treatment group was(57.72±5.66)% for 24 h,(66.72±4.28)% for 48 h and(77.79±10.74)% for 72 h.The inhibition rate of 20umol/L LY294002 treatment group was(68.72±1.3)% for 24 h,(79.92±1.3)% for 48 h and(87.81±1.05)% for 72 h.The inhibition rate of 40umol/L LY294002 treatment group was(71.24±1.63)% for 24 h,(81.05±0,99)% for 48 h and(88.39±0.33)% for 72 h.According to the inhibition rate,IC50 values of LY294002 on SKOV3 cells at different time periods were calculated as follows: IC50 values of 24,48 and 72 were 9.591,7.141 and 5.209umol/L,respectively.IC50(7umol/L)at 48 h was selected for subsequent experiments.2.The results of the cell adhesion experiment showed that: LY294002 can significantly inhibit the adhesion ability of SKOV3 cells,and the cell adhesion rates of the control group and the LY294002 treatment group were(80.00±8.92)% and(61.23±14.18)% respectively,and the difference was statistically significant(P<0.05).3.The cell scratch test results showed that LY294002 could significantly inhibit the migration of SKOV3 cells.After 48 hours of SKOV3 cells were treated with LY294002,the mobility of Skov3 cells was(60.43±1.34)% and(30.46±2.39)%,respectively,and the difference was statistically significant(P < 0.001).4.The results of the Transwell migration experiment showed that the number of migrated cells were(910.67±34.12)and(727.67±27.06)after cultured for 48 hours in the SKOV3 cells of the control group and the SKOV3 cells of the LY294002-treated group,and the difference was statistically significant(P<0.01).Transwell invasion test results: After cultured for 48 hours,the numbers of SKOV3 cells in the control group and SKOV3 cells in the LY294002-treated group were(908.33±6.03)and(518±13.23),respectively,and the difference was statistically significant(P<0.001).5.Western blot results showed that there were no significant differences in the relative expression levels of PI3 K,AKT and m TOR protein in 7umol/L LY294002 treatment group compared with the control group(P>0.05).The relative expression levels of p-PI3 K,p-AKT and p-m TOR in 7umol/L LY294002 treatment group were0.3300±0.0046,0.2220±0.0090 and 0.0490±0.0053,respectively.Compared with the control group of 0.9997±0.0025,0.9407±0.0605 and 0.9403±0.0621,the differences were statistically significant(P < 0.001).Conclusions: 1.At the cellular level,LY294002 has obvious inhibitory effects on the proliferation,adhesion,migration and invasion of ovarian cancer cells.2.The mechanism of LY294002 on the malignant biological behavior of ovarian cancer cells may be related to the PI3K/AKT/m TOR pathway.