目的 研究miR-21在胆管癌组织及细胞系中的表达特征及其在胆管癌发生过程中的功能.方法 用real-time PCR与Northern blot法分析miR-21在胆管癌组织及胆管癌细胞系QBC939中的表达；选择特异抑制剂Anti-miR-21敲低QBC939细胞内源性miR-21的表达,检测细胞增殖及凋亡表型；用萤光素酶双报告基因以及流式细胞仪分析法筛选并鉴定miR-21的靶基因；用体外侵袭实验分析miR-21对胆管癌细胞系QBC939体外侵袭能力的影响.结果 miR-21在胆管癌组织及胆管癌细胞系QBC939中表达均明显上调；QBC939转染Anti-miR-21后,抑制细胞增殖并促进细胞凋亡；miR-21可以抑制RECK的表达,并可以通过两者之间的相互作用,参与QBC939细胞系的体外侵袭调控.结论 抑制miR-21的功能可以抑制胆管癌的进展.%Objective To investigate the expression profile of miR-21 in human cholangiocarcinoma tissues and QBC939 cell line and probe the function of miR-21 in cholangiocarcinogenesis. Methods miR-21 expression in human cholangiocarcinoma tissues and QBC939 cell line was measured by using real-time PCR and Northern Blot, respectively. Cell growth and apoptosis was analyzed in QBC939 after transfection with Anti-miR-21. Specific target analysis was performed by using dual-reporter gene assay and FACS. In Vitro invasion assay was performed to probe the effect of miR-21 on QBC939 invasiveness. Results Expression analysis reveals that miR-21 levels depicted a significant up-regulation as compared to the matched normal bile duct and miR-21 levels were augmented approximately 3. 4-fold in QBC939 cells. Silencing of miR-21 in QBC939 by using Anti-miR-21 decreased cell growth and induced cell apoptosis. RECK was identified as a direct effector of miR-21 and miR-21 promoted QBC939 cell invasion in vitro through negatively regulating miR-21-RECK program. Conclusions Targeting of miR-21 is sufficient to limit cholangiocarcinoma progress.
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