miR125a对胆管癌细胞株QBC939增殖、凋亡影响及可能机制

目的探讨miR125a对胆管癌细胞株QBC939细胞增殖及凋亡的影响及可能机制。方法实时荧光定量PCR检测52例经手术和病理证实的肝外胆管癌,31例癌旁组织及10例正常胆管组织标本miR125a的表达,免疫组化检测MMP-9蛋白表达。miR125a转染胆管癌细胞株QBC939,MTT法检测细胞增殖,流式细胞分析法检测细胞周期,Caspase-3试剂盒检测凋亡,Western blot检测MMP-9蛋白表达。结果胆管癌组织miR125a表达明显下调,低于正常胆管组织(P<0.05),MMP-9明显高于正常胆管组织(P<0.05)。与对照质粒组比较,转染miR125a胆管癌细胞QBC939增殖受到抑制,流式细胞分析法表明转染miR125a的胆管癌细胞QBC939导致G1阻滞,Caspase-3试剂盒检测显示凋亡增加,Western blot分析表明转染miR125a的胆管癌细胞QBC939中MMP-9的表达出现明显的下调。结论miR125a在肝外胆管癌中低表达,它们可能与胆管癌的发生有关。胆管癌细胞株QBC939 miR125a表达上调,能够抑制细胞增殖,诱导细胞凋亡和G1阻滞,miR125a的作用可能与MMP-9的表达相关。

Effects of miR125a on cell cycle and apoptosis of cholangiocarcinoma cell line QBC939 and its mechanism

Aim To explore the effects of miR125a on cell cycle and apoptosis inducing of cholangiocarcinoma cell line QBC939 and its mechanism.Methods The expression of miR125a was detected by real-time PCR on 52 specimens of extrahepatic cholangiocarcinoma and 10 normal bile duct.MMP-9 was assayed by immunohistochemistry.Expression plasmid of miR125a was transiently transfected into human cholangiocarcinoma cell line QBC939 and then proliferation was measured by MTT method.Cell cycle was analyzed with flow cytometry.MMP-9 expression was measured by Western blot.Results Compared with normal bile duct,the relative expression ratio of miR125a in extrahepatic cholangiocarcinoma was decreased and MMP-9 expression was up-regulated.Compared to control group,the proliferation of miR125a transfected cholangiocarcinoma cell was effectively inhibited and miR125a induced G1 phase arrest.MMP-9 expression was increased in miR125a transfected cholangiocarcinoma cells.Conclusions The expression of miR125a is down-regulated in extrahepatic cholangiocarcinoma,and it may be correlated with tumorigenesis.Up-regulation of miR125a in cholangiocarcinoma cell line QBC939 inhibits cell proliferation,arrests cell cycle which may be associated with MMP-9 expression.