circ_0000592/miR-515-5p轴调控乳腺癌细胞增殖、迁移和侵袭

    Regulation of circ_0000592/miR-515-5p axis in the proliferation, migration and invasion of breast cancer cells

    • 摘要:
      目的 探讨circ_0000592/miR-515-5p轴对乳腺癌细胞增殖、迁移和侵袭的影响。
      方法 收集57例乳腺癌病人的癌组织和癌旁组织, qRT-PCR法检测组织中circ_0000592和miR-515-5p表达;体外培养人乳腺上皮细胞HMEpiC和乳腺癌MCF-7细胞, qRT-PCR法检测细胞中circ_0000592和miR-515-5p表达。双荧光素酶报告基因实验验证circ_0000592和miR-515-5p的调控关系。转染circ_0000592小干扰RNA、或共转染circ_0000592小干扰RNA与miR-515-5p抑制剂至MCF-7细胞, CCK-8实验、克隆形成实验、Transwell分别检测细胞存活率、克隆数、迁移数和侵袭数, 蛋白质印迹法检测细胞中E-cadherin和N-cadherin蛋白表达。
      结果 乳腺癌组织中circ_0000592表达升高(P < 0.05), miR-515-5p表达降低(P < 0.05)。乳腺癌MCF-7细胞中circ_0000592表达较HMEpiC细胞升高(P < 0.05), miR-515-5p表达较HMEpiC细胞降低(P < 0.05)。circ_0000592在MCF-7细胞中靶向结合并负调控miR-515-5p。下调circ_0000592后, MCF-7细胞存活率、克隆数、迁移数、侵袭数及细胞中N-cadherin蛋白表达均降低(P < 0.01), E-cadherin蛋白表达升高(P < 0.01)。下调miR-515-5p部分减弱下调circ_0000592对MCF-7细胞增殖、迁移和侵袭的作用。
      结论 circ_0000592在乳腺癌组织及细胞中表达升高, 其通过靶向负调控miR-515-5p促进乳腺癌细胞增殖、迁移和侵袭。

       

      Abstract:
      Objective To investigate the effect of circ_0000592/miR-515-5p axis on the proliferation, migration and invasion of breast cancer cells.
      Methods The cancer tissues and adjacent tissues of 57 breast cancer patients were collected, and the expressions of circ_0000592 and miR-515-5p in the tissues were detected by qRT-PCR. Human breast epithelial cells HMEpiC and breast cancer MCF-7 cells were cultured in vitro. qRT- PCR method was used to detect the expression of circ_0000592 and miR-515-5p in the cells. The dual luciferase reporter gene experiment verified the regulatory relationship between circ_0000592 and miR-515-5p. MCF-7 cells were transfected with circ_0000592 small interfering RNA or miR-515-5p inhibitor, and then CCK-8 assay, clone formation test and Transwell assay were used to detect the cell viability, clone number, migration number and invasion number. The protein expression of E-cadherin and N-cadherin in cells were detected by Western blotting.
      Results In breast cancer tissues, the expression of circ_0000592 was increased (P < 0.05), and the expression of miR-515-5p was decreased (P < 0.05). The expression of circ_0000592 in breast cancer MCF-7 cells was higher than that in HMEpiC cells (P < 0.05), but the expression of miR-515-5p was lower than that in HMEpiC cells (P < 0.05). Circ_0000592 could negatively regulate miR-515-5p in MCF-7 cells. After down-regulating circ_0000592, the survival rate, clone number, migration number, invasion number of MCF-7 cells and the expression of N-cadherin protein in the cells were all decreased (P < 0.01), but the expression of E-cadherin protein was increased (P < 0.01). Down-regulation of miR-515-5p partially restored the effectd of down-regulation of circ_0000592 on proliferation, migration and invasion of MCF-7 cells.
      Conclusions The expression of circ_0000592 is elevated in breast cancer tissues and cell lines, which promotes breast cancer cell proliferation, migration and invasion through the negative regulation of miR-515-5p.

       

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