The Experimental Study Of The Influence And Mechanism Of Ellipticine On Human Bladder Cancer

BackgroundIn our country,the incidence of bladder cancer ranks first in the urinary tract tumors.In recent years,the incidence of bladder cancer still presents a rising trend year by year.The main treatments of bladder cancer are surgery and chemotherapy,etc.Ellipticine is a kind of natural alkaloids,and was proved to have tumor inhibitory role in a wide variety of tumors,but its effect and mechanism of bladder cancer remain unclear.PurposeThis study aims to explore the effects and the mechanisms of ellipticine on cell proliferation,cell cycle and movement ability in T24 bladder cancer by in vitro experiments.MethodsT24 cells were treated with different concentrations(1-16 ?M)of ellipticine and in different time(24-72 hours).MTT assay and colony forming assay were used to detect the change of cell proliferation and viability.Flow cytometry was used to detect the change of cell cycle.Transwell experiment was taken to test the change of cell movement ability.The expression of MMP and p53 mRNA were measured by RT-PCR.The expression of the corresponding proteins was tested by Western Blot experiments.ResultsEllipticine inhibited the proliferation of T24 bladder cancer cell in a time-dose dependent way.With the concentration higher and the treating time longer,the inhibition ability got stronger.Ellipticine induced a G2/M arrest,upregulated the expression of cyclin B1 and downregulated the expression of cdkl.Ellipticine increased the expression of both p53 mRNA and protein.Ellipticine inhibited the migration of T24 cells,decreased the expression of MMP-9 and increased the expression of E-cadherin.Ellipticine increased ATM,Cdc25C(Ser-216)and Chkl(Ser-345)phosphorylation level.After the knockdown of ATM by siRNA interference,the changes of Cdc25C(Ser216)and Chkl(Ser-345)were reversed.ConclusionEllipticine inhibits cell proliferation,induces G2/M phase cell cycle arrest and inhibits migration ability of T24 cells in a dose-time dependent way.The inhibition of ellipticine on T24 cell was induced by the activation of ATM-Chkl-Cdc25C-Cdkl and p53 pathway and the regulation of the EMT and MMPs.