Effects Of PP2A On The Biological Function Of Endometrial Cancer Ishikawa Cells

Objective: To investigate the effects of protein phosphatase 2A(PP2A)specific inhibitors Okadaic acid(OA)and agonists D-erythrosphingosine(DES)on endometrial cancer cell lines(Ishikawa cells),and their effects on the expression of PP2 A at the cellular level and cell function.Method: 1.Ishikawa cells were routinely cultured,and cells in logarithmic growth phase were taken using PP2 A agonists(DES concentrations of 0nmol/l,5nmol/l,10nmol/l,20nmol/l)and PP2 A inhibitors(OA concentrations of 0nmol/l,10nmol/l,20nmol/l,40nmol/l)after treatment for 12 h,24h,48 h,respectively,add CCK8 solution to detect the proliferation and inhibition of cells by OA and DES;2.Further select the concentration and time point where the proliferation and inhibition rates of Ishikawa cells are most obvious as the experimental group,and set a control group,where the concentration of DES is 20nmol/l,and the concentration of OA is 40nmol/l.Scratch test was used to detect cell migration ability,Treanswell test was used to detect cell invasion ability,flow cytometry was used to measure cell cycle,and Western Blot method was used to detect cell-level PP2 A expression changes.Result: 1.DES showed an inhibitory effect on Ishikswa cells as a whole;comparing them in pairs,it was found that as the concentration and time increased,the inhibition rate showed a time-dose dependence;of which DES concentration was 20 n M,At 48 h,its inhibitory effect was the most obvious.OA generally showed a pro-proliferative effect on Ishikswa cells.Comparisons were performed in pairs,and the proliferation rate showed a time-dose dependence with the increase of time and concentration.Among them,the concentration of OA was 40 n M and the time was 48 h.The most obvious proliferation effect.2.When the concentration of OA of 40nmol/l and DES of 20nmol/l were applied to Ishikawa cells for 48 h,the scratch test showed that compared with the control group,OA can significantly promote cell migration,and DES can significantly inhibit cell migration.Difference All were statistically significant.The Treanswell test showed that compared with the control group,OA significantly promoted cell invasion,and DES significantly inhibited cell invasion,and the differences were statistically significant.3.When OA at a concentration of 40nmol/l and DES at 20nmol/l were applied to Ishikawa cells for 48 hours,the flow cytometry cycle test showed that the proliferation activity of the cells in the OA group was stronger than that in the control group,and the difference was statistically significant.The proliferation activity of the cells in the DES group was lower than that of the control group,and the difference was statistically significant.4.When OA at a concentration of 40nmol/l and DES at 20nmol/l were applied to cells for 48 hours,Western Blot analysis showed that compared with the control group,OA could inhibit the expression of PP2 A protein at the cellular level,and DES could promote the expression of PP2 A protein at the cellular level.Conclusion: 1.OA and DES can change the expression of PP2 A in human endometrial cancer Ishikawa cells.OA inhibits the expression of PP2 A,and DES promotes the expression of PP2 A.2.OA and DES,as commonly used inhibitors and agonists of exogenous PP2 A,affect the biological function of Ishikawa cells by changing the expression of PP2 A at the cellular level,including cell proliferation,migration,invasion and cell cycle.