摘要目的 探讨微小RNA(miRNA,miR)-148b和miR-152在胆管癌中的表达及其生物学功能.方法 用实时定量聚合酶链反应(Real-time PCR)检测miR-148b和miR-152在胆管癌与正常胆管组织、胆管癌细胞株QBC939和良性胆管细胞株H-69中的表达;分别用Real-time PCR、噻唑蓝(MTT)检测法、克隆形成实验和流式细胞术检测miR-148b和miR-152的表达及其对细胞增殖、细胞周期和凋亡的影响.结果 与正常胆管组织比较,miR-148b和miR-152在胆管癌组织中分别下调(2.75±0.16)倍和(3.05 ±0.18)倍,差异均有统计学意义(P=0.013、0.019);与良性胆管细胞比较,miR-148b和miR-152在胆管癌细胞中分别下调(2.52±0.03)倍和(3.96±0.02)倍,差异均有统计学意义(P=0.018、0.012);miR-148b和miR-152过表达显著抑制胆管癌细胞的增殖速率,分别为(38.49±0.31)％和(52.69±1.01)％,同时也能显著抑制克隆形成率(P=0.031、0.023),miR-148b和miR-152均可使细胞周期阻滞于S期(P=0.027、0.017),并且miR-148b和miR-152过表达均可诱导细胞凋亡.结论 胆管癌组织中miR-148b和miR-152表达均下调,过表达miR-148b和miR-152后可抑制胆管癌细胞增殖及诱导胆管癌细胞凋亡.

abstractsObjective To investigate the microRNA (miRNA,miR)-148b and miR-152 expression and their biological functions in cholangiocarcinoma cells.Methods The expression levels of miR-148b and miR-152 in cholangiocarcinoma simples and QBC939 cells were detected using real-time quantitative polymerase chain reaction (Real-time PCR).After transfection with miR-148b/ miR-152 mimics,cell proliferation,cell cycle and apoptosis were assessed using methyl thiazol tetrazolium(MTT) method,clone formation test and flow cytometry,respectively.Results The expression levels of miR-148b and miR-152 were significantly down-regulated [(2.75 ± 0.16) folds and (3.05 ± 0.18) folds] in cholangiocarcinoma samples as compared with normal samples (P =0.013,0.019,respectively),and [(2.52 ±0.03) folds and (3.96 ±0.02) folds] in QBC939 cells as compared with H-69 cells (P =0.018,0.012,respectively).Overexpression of miR-148b or miR-152 could significantly inhibit cell proliferation [(38.49 ± 0.31) ％ and (52.69 ± 1.01) ％,respectively],and suppress cell clone formation (P =0.027,0.017,respectively),block cell cycle in the S phase and induce cell apoptosis.Conclusion MiR-148b and miR-152 were down-regulated in cholangiocarcinoma cells,and overexpression of miR-148b and miR-152 could inhibit cell proliferation and induce cell apoptosis.