溴苯基姜黄素对胆管癌细胞凋亡、迁移和侵袭的影响及机制研究

目的:研究溴苯基姜黄素(GL63)对人胆管癌RBE细胞凋亡、迁移和侵袭的影响,并基于Janus激酶(JAK)/信号转导及转录激活因子(STAT)信号通路探讨其作用机制.方法:采用MTT法检测不同浓度GL630(空白对照,下同)、1.25、2.5、5、10、20、40μmol/L]作用48 h对RBE细胞增殖的影响,并...

Study on the Effect and Its Mechanism of Bromophenylcurcumin on Apoptosis,Migration and Invasion of Cholangiocarcinoma Cells

OBJECTIVE:To study the effects of bromophenylcurcumin(GL63)on the apoptosis,migration and invasion of human cholangiocarcinoma RBE cells,and to investigate its mechanism based on JAK/STAT signaling pathway.METHODS:MTT assay was used to detect the effects of different concentrations of GL630(blank control,similarly hereinafter),1.25,2.5,5,10,20,40μmol/L]on the proliferation of RBE cells after 48 h treatment;the IC₅₀ was calculated.The effects of different concentrations of GL63(0,5,10,20μmol/L)on colony formation were detected by crystal violet staining after 48 h treatment.Flow cytometry,Hoechst 33342 staining,cell scratch test and Transwell chamber invasion test were used to detect the effects of different concentrations of GL63(0,5,10,20μmol/L)on cell cycle distribution,apoptosis,migration and invasion ability after 24 h treatment.Western blotting assay was adopted to detect the effects of different concentration of GL63(0,5,10,20μmol/L)on the expression of JAK2/STAT3 signal pathway associated proteins.RESULTS:The proliferation inhibition rates of RBE cells in different concentrations of GL63 groups(1.25-40μmol/L)were significantly increased,compared with blank control group(P<0.01),and showed a dose-dependent trend,with IC₅₀ of(8.46±1.30)μmol/L.Compared with blank control group,inhibition rates of RBE cell colony formation were significantly decreased in different concentrations(5,10,20μmol/L)of GL63 groups(P<0.01).The percentage of RBE cells at G0/G1 phase increased significantly,while that at S phase decreased significantly(P<0.01).The apoptotic rate increased significantly(P<0.01),and the nucleus showed dense pyknosis and apoptotic bodies.The rate of cell migration and healing was significantly decreased(P<0.01),and the number of invasive cells through basement membrane was significantly decreased(P<0.01).The protein expression of p-JAK2,p-STAT3,Bcl-2,MMP-2,MMP-9,Pro-caspase-9 and Pro-caspase-3 were down-regulated significantly while the expression of Bax,Cyt-c,Cleaved-caspase-9 and Cleaved-caspase-3 were up-regulated significantly(P<0.01).CONCLUSIONS:GL63 may inhibit the proliferation,migration and invasion of RBE cells and promote its apoptosis by inhibiting JAK2/STAT3 signal pathway.