Effect And Mechanism Of Arsenic Trioxide On Invasion And Metastasis Of Human Cholangiocarcinoma Cell

AIM:To investigate the effect of arsenic trioxide (As₂O₃) on heterogeneous adhesion,invasion and expression of urokinase-type plasminogen activator,matrix metalloprotease-9 in human cholangiocarcinoma cells in vitro. In orde to provide the theoretical basis for application of As₂O₃ in therapy of the cholangiocarcinoma.METHODS:1 Cholangiocarcinoma cells (QBC-939)were cultured regularly,experimental group treated with 4μmol/L As₂O₃,control group treated with equal volume RPMI-1640,and set up a blank group.After 37℃cultured 72 h, heterogeneous adherence ability of QBC-939 were assessed by MTT in different time points(30min,60 min,90 min,120 min).2 The control group (only RPMI-1640 added) and different concentration group of As₂O₃ (1,2,4,8μmol / L) were seeded in Transwell chamber separately,and detecting cell invasion ability after cultured 72h.3 After cholangiocarcinoma cells were treated for 72h with different concentrations of As₂O₃ (1,2,4,8μmol / L),and treated with 4μmol/ L As₂O₃ for different times(24,48,72,96 h),Reverse transcription-polymerase chain reaction (RT-PCR) was employed to measure the expression of u-PA,MMP-9 mRNA.Results:1 Cholangiocarcinoma cells were treated with 4μmol/L As₂O₃ for72h,the adherence rate in 30,60,90,120 min of experimental group were 82.4%,79%,75.2%,71.4%, the adherence rate of 120min was the lowest(71%), each experimental group compared with the corresponding control group(100%)P < 0. 05,there is no signifieant difference between 30min and 60min(P> 0. 05).2 After cholangiocarcinoma cells were treated with 2,4,8μmol/L As₂O₃ for 72h,the number of cholangiocarcinoma cells with invasion were(70±9),(37±6),(21±4).Compared with the control group(97±7)P < 0. 05.With the increase of As₂O₃ concentration the number of invasion was reduced gradually. 3 After different coneentration of As₂O₃ treated for 72h,with the increase of As₂O₃ concentration the expression level of uPA and MMP-9mRNA was reduced gradually,the apropriate concentration of As₂O₃ was 4μmol/ L.After 4μmol/ L As₂O₃ treated for different times,with the increase of time the expression level of uPA and MMP-9mRNA was reduced gradually,with maximal reduce at 72h.Conclusion:The adherence ability of cholangiocarcinoma cells was efficiently inhibited by As₂O₃ at definite coneentration(4μmol/ L).As₂O₃ can inhibit the capability of invasion in a dose-dependent manner.As₂O₃ can inhibit the expression of u-PA,MMP-9 mRNA in a time and dose-depended manner,while the significant inhibition at 4μmol/ L As₂O₃ treated 72h.As₂O₃ has the effects of antagonizing cholangiocarcinoma cell invasion and migration in vitro,the mechanism may be involved in down-regulating the expressions of u-PA and MMP-9 mRNA in cholangiocarcinoma cells.