circ-0005273靶向miR-1200对食管癌Eca109细胞增殖和凋亡的影响

Effects of circ-0005273 targeting miR-1200 on the proliferation and apoptosis of esophageal cancer Eca109 cells

  • 摘要:
      目的  探讨circ-0005273对食管癌细胞增殖和凋亡的影响及可能机制。
      方法  收集41例食管癌患者的癌组织及癌旁组织,采用实时荧光定量聚合酶链式反应(RT-qPCR)检测组织中circ-0005273和miR-1200表达。体外培养食管癌Eca109细胞,分别转染circ-0005273小干扰RNA、miR-1200模拟物,或共转染circ-0005273小干扰RNA与miR-1200抑制剂。采用CCK-8法和克隆形成实验检测细胞增殖;流式细胞术检测细胞凋亡;蛋白印迹法检测细胞中cleaved-caspase9、cleaved-caspase3蛋白表达;双荧光素酶报告基因实验验证miR-1200与circ-0005273调控关系。
      结果  食管癌组织中circ-0005273的表达量高于癌旁组织,而miR-1200的表达量低于癌旁组织,差异均有统计学意义(P<0.05)。干扰circ-0005273表达或过表达miR-1200后,Eca109细胞光密度(OD)值和克隆形成数降低,凋亡率和cleavde-caspase9、cleavde-caspase3蛋白的表达量增加,差异均有统计学意义(P<0.05)。circ-0005273靶向负调控miR-1200,干扰miR-1200可逆转干扰circ-0005273对Eca109细胞增殖和凋亡的影响。
      结论  circ-0005273在食管癌组织中呈高表达,干扰circ-0005273可能通过靶向负调控miR-1200阻碍了食管癌细胞增殖,并加剧了细胞凋亡。

     

    Abstract:
      Objective  To investigate the effects of circ-0005273 on the proliferation and apoptosis of esophageal cancer cells and its possible mechanism.
      Methods  The cancer tissues and adjacent tissues of 41 patients with esophageal cancer were collected, and the expression of circ-0005273 and miR-1200 in the tissues were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). Esophageal cancer Eca109 cells were cultured in vitro, and were respectively transfected with circ-0005273 small interfering RNA (si-circ-0005273) or miR-1200 mimic, or co-transfected with circ-0005273 small interfering RNA and miR-1200 inhibitor. CCK-8 method and clone formation experiment were used to detect cell proliferation, flow cytometry was used to detect cell apoptosis, the western blotting was used to detect the protein expression of cleaved-caspase9 and cleaved-caspase3 in cells, and the dual luciferase reporter gene experiment was used to verify the regulatory relationship between miR-1200 and circ-0005273.
      Results  The expression of circ-0005273 in esophageal cancer tissues was significantly higher than that in adjacent tissues, but the expression of miR-1200 was significantly lower than that in adjacent tissues (P < 0.05). After disturbing circ-0005273 or over-expressing miR-1200, the optical density (OD) value and clone formation number of Eca109 cells significantly decreased (P < 0.05), but the apoptosis rate and the protein expression of cleavde-caspase9 and cleavde-caspase3 increased significantly (P < 0.05). Circ-0005273 was able to target and negatively regulate miR-1200, and disturbing miR-1200 reversed the effect of disturbing circ-0005273 on the proliferation and apoptosis of Eca109 cells.
      Conclusion  Circ-0005273 is highly expressed in esophageal cancer tissues. Disturbing circ-0005273 may hinder the proliferation of esophageal cancer cells and aggravate cell apoptosis by targeting and negatively regulating miR-1200.

     

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