MicroRNA-133表达对人胆管癌细胞QBC939迁移和侵袭的影响

Effect of microRNA-133 on migration and invasion of cholangiocarcinoma cells

摘要:

目的 探讨MicroRNA-133（miR-133）表达对人胆管癌细胞QBC939迁移、侵袭的影响，探寻其中可能存在的分子机制。方法 实时荧光定量聚合酶链反应（qRT-PCR）检测人胆管癌细胞QBC939 miR-133各亚型的表达，选择适合的亚型进行实验。将人胆管癌细胞QBC939分为3组：干扰组（转染miR-133a-5p抑制物），阴性对照组（转染miR-133a-5p模拟物）及空白对照组，qRT-PCR检测3组miR-133a-5p mRNA相对表达量；Transwell实验检测各组人胆管癌细胞迁移和侵袭的能力；Western blotting检测3组人胆管癌细胞中LASP-1蛋白家族成员（LIM、SH3-1蛋白）的相对表达量。结果 miR-133的3种亚型中，miR-133a-5p mRNA相对表达量最高（P <0.05）。干扰组人胆管癌细胞的miR-133a-5p mRNA相对表达量（0.70±0.08）低于阴性对照组及空白对照组（P <0.05）。与阴性对照组及空白对照组比较，干扰组细胞迁移数（72.0±11.0）和侵袭数（20.0±3.0）明显减少（P <0.05）。与阴性对照组及空白对照组比较，干扰组人胆管癌细胞的LIM蛋白相对表达量最高，SH3-1蛋白相对表达量最低（P <0.05）。结论 miR-133a-5p能调控LIM蛋白、SH3-1蛋白的表达。miR-133a-5p低表达能抑制人胆管癌细胞QBC939的迁移和侵袭，这可能是通过miR-133a-5p对LASP-1蛋白表达的调控实现的。miR-133a-5p有望成为胆管癌靶向治疗的新靶点。

Abstract:

Objective To study the expression of MicroRNA-133 (miR-133) in QBC939 cholangiocarcinoma cells, to analyze the role of miR-133 in the migration and invasion of cholangiocarcinoma cells, and to explore the possible molecular mechanisms.Methods RT-PCR (real-time PCR) was used to analyze the expression of each subtype of miR-133 in QBC939 cholangiocarcinoma cells in order to select the appropriate subtype for functional experiments. QBC939 cells were divided into three groups: interference group (transfection with miR-133a-5p inhibitor), negative control group (transfection with miR-133a-5p mimic), and blank control group. RT-PCR was used to measure the expression of miR-133a-5p in each group; Transwell assay was used to analyze the migration and invasion of cholangiocarcinoma cells in each group; Western blotting was used to detect the expression of LASP-1 protein family members (LIM, SH3-1 protein) in each group.Results In the three subtypes of miR-133，the relative expression of miR-133a-5p was highest (P < 0.05). The relative expression of mir-133a-5p in the interference group was (0.70 ± 0.08), which was lower than that in the negative control group and blank control group (P < 0.05). Compared with the negative and blank control groups, the number of cell migration (72.0 ± 11.0) and invasion (20.0 ± 3.0) in the interference group was significantly decreased (P < 0.05). Compared with the negative and blank control groups, the interference group exhibited higher expression level of LIM protein and lower expression level of SH3-1 protein (P < 0.05).Conclusion MiR-133a-5p can regulate the expression of LASP-1 protein (LIM protein, SH3-1 protein) in QBC939 cholangiocarcinoma cells. The low expression of miR-133a-5p can inhibit the migration and invasion of QBC939 cells, which may be achieved through the regulation of LASP-1 protein by miR-133a-5p. Mir-133a-5p is expected to become a new target for targeted therapy of cholangiocarcinoma.