Objective: To investigate the expression of miR-29b in cholangiocarcinoma and explore its effects on cell proliferation and apoptosis of cholangiocarcinoma cells.
Methods: Real-time PCR was used to detect the expression of miR-29b in cholangiocarcinoma cells line QBC939 and cholangiocarcinoma tissues. The lentiviral vector LV-hsa-miR-29b and blank vector were constructed to infect QBC939 cells. MTT assay and cell clone formation assay were performed to assess the changes in the cell proliferation and clone formation, respectively; flow cytometry was employed to evaluate the effect of miR-29b overexpression on cell cycle and apoptosis.
Results: The expression of miR-29b was significantly down-regulated in QBC939 cells and cholangiocarcinoma tissues as compared with H-69 cells and normal tissues (P < 0.01). Compared with the blank vector, the lentiviral vector LV-hsa-miR-29b caused significantly increased expression of miR-29b in QBC939 cells (P < 0.01), which exhibited suppressed cell proliferation and clone formation (P < 0.01 or 0.05), cell cycle arrest at the S phase (P < 0.05), and significantly increased cell apoptosis (P < 0.01).
Conclusions: As a tumor-suppressing miRNA, miR-29b is down-regulated in cholangiocarcinoma, and its overexpression can suppress the proliferation and induce apoptosis of cholangiocarcinoma cells.
目的: 研究miR-29b在胆管癌中的表达情况，及其对胆管癌细胞QBC939增殖和凋亡的影响。
方法: Real-time PCR检测miR-29b在胆管癌细胞QBC939与良性胆管细胞株H-69中的表达；及胆管癌组织与正常胆管组织中的表达。将对数生长期的QBC939细胞分为3组，空白对照组，阴性对照组，miR-29b过表达组。MTT和细胞克隆形成实验分别检测过表达miR-29b对细胞增殖和克隆形成率的影响；流式细胞术检测过表达miR-29b对细胞周期和凋亡的影响。
结果: miR-29b在胆管癌细胞及胆管 癌组织中表达较正常胆管细胞和组织中均显著降低（P < 0.01）。miR-29b过表达组QBC939细胞的miR-29b较阴性对照组表达明显升高（P < 0.01）。在QBC939细胞中，过表达miR-29b可以显著抑制细胞的增殖和克隆形成（P < 0.01和P < 0.05）；过表达miR- 29b可阻滞细胞在S期（P < 0.05），并且导致细胞凋亡明显增加（P < 0.01）。
结论: miR-29b作为一种抑癌microRNA，在胆管癌中低表达，过表达miR-29b可以抑制胆管癌细胞增殖，促进细胞凋亡。
Keywords: apoptosis; cholangiocarcinoma; miR-29b; proliferation.