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目的研究miR-125b在乳腺癌中的表达,尤其是其对乳腺癌迁移侵袭的作用以及该过程所涉及的分子机制。方法 (1)用实时荧光定量PCR检测23对临床组织样本中miR-125b及靶基因ETV6的表达水平;(2)用实时荧光定量PCR、双荧光报告验证靶基因;(3)在细胞中瞬时转染miR-125b mimics,并用划痕、transwell迁移、侵袭及Western blot检测体外乳腺癌细胞的迁移侵袭能力及上皮间质转化的变化;(4)用靶基因敲减实验进一步验证靶基因的准确性。结果 (1)与正常组织相比,乳腺癌组织中miR-125b的表达显著降低(P<0.001),ETV6的表达显著增高(P<0.05);(2)ETV6为miR-125b的靶基因;(3)与对照相比,上调的miR-125b可显著抑制Hs578T的迁移和侵袭(P<0.001),并抑制EMT的发生;(4)敲减ETV6对Hs578T的作用与miR-125b过表达一致。结论 miR-125b在乳腺癌中低表达,miR-125b通过靶向结合ETV6 3’UTR抑制该基因的表达,从而抑制Hs578T的迁移、侵袭及上皮间质转化的发生。因此,miR-125b可作为抑癌基因发挥作用。
Objective To investigate the expression of miR-125b in breast cancer, especially its role in the invasion and metastasis of breast cancer and the molecular mechanisms involved in this process. Methods (1) Real-time fluorescence quantitative PCR was used to detect the expression levels of miR-125b and target gene ETV6 in 23 clinical tissue samples; (2) Real-time fluorescence quantitative PCR and double fluorescence reporter were used to verify the target genes; (3) MiR-125b mimics were transfected and the invasion, migration and invasion of breast cancer cells in vitro were detected by scratch, transwell migration, invasion and Western blot. (4) The target gene knockdown was used to further verify the accuracy of the target gene Sex. Results (1) The expression of miR-125b in breast cancer tissues was significantly lower than that in normal tissues (P <0.001), and the expression of ETV6 was significantly increased (P <0.05). (2) ETV6 was the target gene of miR- (3) Compared with the control, up-regulated miR-125b significantly inhibited the migration and invasion of Hs578T (P <0.001) and inhibited the occurrence of EMT. (4) The effect of knockdown of ETV6 on Hs578T was consistent with the overexpression of miR-125b . Conclusion miR-125b is down-regulated in breast cancer. MiR-125b inhibits the migration, invasion and epithelial-mesenchymal transition of Hs578T by targeting the ETV6 3’UTR. Therefore, miR-125b can function as a tumor suppressor gene.