Sun Lei, Wang Hua, Jiang Jue, Bi Xueyuan
Department of Ultrasound Laboratory, Second Affiliated Hospital, Xi'an Jiaotong University, Xi'an 710004, China.
Department of Pharmacy, Honghui Hospital Affiliated to Xi'an Jiaotong University, Xi'an 710054, China. *Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2020 Feb;36(2):138-144.
Objective To investigate the molecular mechanism of miR-153-3p regulating the proliferation, migration and invasion of human breast cancer cells. Methods Bioinformatics software was used to predict the candidate target gene of miR-153-3p. The double-luciferase reporter assay was used to validate the targeting relationship between miR-153-3p and candidate genes. Real-time fluorescence quantitative PCR and Western blotting were performed to detect the effect of miR-153-3p on the mRNA and protein expressions, respectively. The cell proliferation ability was examined by MTT assay. The migration ability of cells was determined in a scratch assay. The invasion ability was detected by Transwell assay. Results Software-predicted results showed a continuous binding region between miR-153-3p and Rho-associated coiled-coil containing protein kinase 1 (ROCK1). The miR-153-3p mimics co-transfected with ROCK1 wild-type reporter vector significantly decreased the relative activity of luciferase in MDA-MB-231 cells. The transfection of hsa-miR-153-3p mimics increased miR-153-3p expression and decreased ROCK1 protein expression. In comparison with the control cells, the proliferation, migration and invasion of MDA-MB-231 cells over-expressing miR-153-3p were dramatically inhibited. Conclusion The miR-153-3p can inhibit the proliferation, migration and invasion of breast cancer cells via down-regulation of ROCK1.
目的 探讨miR-153-3p调控人乳腺癌细胞增殖、迁移及侵袭的分子机制。方法 运用生物信息学软件预测miR-153-3p的候选靶基因。采用双荧光素酶报告基因检测法验证miR-153-3p与候选基因的靶向关系。分别通过实时荧光定量PCR和蛋白质免疫印迹法检测miR-153-3p对mRNA和蛋白表达的影响。采用MTT法检测细胞增殖能力。通过划痕实验测定细胞迁移能力。运用Transwell实验检测侵袭能力。结果 软件预测结果显示miR-153-3p与含Rho相关卷曲螺旋蛋白激酶1（ROCK1）之间存在连续结合区域。miR-153-3p模拟物与ROCK1野生型报告载体共转染后显著降低了MDA-MB-231细胞中荧光素酶的相对活性。转染hsa-miR-153-3p模拟物可增加miR-153-3p表达并降低ROCK1蛋白表达。与对照细胞相比，过表达miR-153-3p的MDA-MB-231细胞的增殖、迁移及侵袭能力均受到显著抑制。结论 miR-153-3p可通过下调ROCK1抑制乳腺癌细胞的增殖、迁移及侵袭。
