南方医科大学学报

南方医科大学学报 ›› 2019, Vol. 39 ›› Issue (02): 144-.doi: 10.12122/j.issn.1673-4254.2019.02.03

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miR-593 通过调控PLK1基因的表达抑制结肠癌细胞的增殖

马金珠，朱益平，王箴，昝嘉伟，曹陇，冯遵永，王森林，范倩，颜亮   

  • 出版日期:2019-02-20 发布日期:2019-02-20

miR-593 inhibits proliferation of colon cancer cells in vitro by down-regulating PLK1

  • Online:2019-02-20 Published:2019-02-20

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摘要： 目的探讨miR-593在结肠癌细胞增殖中的作用及分子机制。方法利用生物信息学分析筛选miR-593可能结合的靶基 因为PLK1；利用双荧光素酶报告基因实验验证miR-593 和PLK1 基因的结合；通过qRT-PCR、Western blotting 验证PLK1 为 miR-593直接作用的靶基因；通过CCK-8实验证明miR-593通过调控PLK1基因的表达抑制结肠癌细胞的增殖。结果运用三 种生物信息学软件对PLK1可能结合的miRNA进行了预测与分析，发现miR-593可能结合PLK1基因；双荧光素酶报告基因实 验证明miR-593 与PLK1 基因发生了结合；Western blotting 结果表明，PLK1 在miR-593 mimic 转染组表达量显著下降，而在 miR-593 inhibitor 组明显升高，相对于对照组均具有统计学差异（P<0.05）；qRT-PCR结果表明，PLK1 RNA水平在各组中表 达水平差异无统计学意义（P>0.05）；细胞增殖实验表明，miR-593与PLK1对结肠癌细胞增殖的影响为相反关系，且共转染miR-593 mimic与PLK1过表达质粒组对细胞增殖的影响介于单独转染miR-593 mimic组与PLK1过表达质粒组之间。结论miR-593通 过调控PLK1基因的表达抑制了结肠癌细胞的增殖，并发挥抑癌miRNA的作用。

Abstract: Objective To explore the role of miR-593 in regulating the proliferation of colon cancer cells and the molecular mechanism. Methods Bioinformatics analysis identified PLK1 as the possible target gene of miR-593. Luciferase assay was employed to verify the binding between miR-593 and PLK1, and qRT-PCR and Western blotting were used to verify that PLK1 was the direct target gene of miR-593. CCK-8 assay was performed to test the hypothesis that miR-593 inhibited the proliferation of colon cancer cells by targeting PLK1. Results Luciferase assay identified the specific site of miR-593 binding with PLK1. Western blotting showed a significantly decreased expression of PLK1 in the colon cancer cells transfected with miR-593 mimics and an increased PLK1 expression in the cells transfected with the miR-593 inhibitor as compared with the control cells (P<0.05). The results of qRT-PCR showed no significant differences in the expression levels of PLK1 among the cells with different treatments (P>0.05). The cell proliferation assay showed opposite effects of miR-593 and PLK1 on the proliferation of colon cancer cells, and the effect of co-transfection with miR-593 mimic and a PLK1-overexpressing plasmid on the cell proliferation was between those in PLK1 over-expressing group and miR-593 mimic group. Conclusion miR-593 inhibits the proliferation of colon cancer cells by down-regulating PLK1 and plays the role as a tumor suppressor in colon cancer.