microRNA-574-3p 抑制结肠癌细胞增殖的机制研究
作者:
作者单位:

1.石家庄市中医院普外科,石家庄 050051;2.秦皇岛市第一医院普外科,河北 秦皇岛 066000

中图分类号:

R-33


Mechanism of microRNA-574-3p in inhibiting the proliferation of colon cancer cells
Author:
Affiliation:

1.Department of General Surgery, Shijiazhuang Hospital of Traditional Chinese Medicine, Shijiazhuang 050051, China. 2. Department of General Surgery, First Hospital of Qinhuangdao, Qinhuangdao 066000

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    摘要:

    目的 观察微小 RNA-574-3p(microRNA-574-3p,miR-574-3p)对结肠癌细胞增殖的影响,并探讨其潜在的作用机制。 方法 采用结肠癌细胞 HCT116 作为实验对象,细胞转染 miR-574-3p mimic 和阴性对照分别作为 miR-574-3p mimic 组和阴性对照组,同时以正常 HCT116 细胞作为空白对照组。 RT-qPCR 检测细胞 miR-574-3p 表达;CCK-8 法检测细胞增殖活力;平板克隆实验检测细胞克隆形成能力;流式细胞术检测细胞周期变化;Western blot 法检测 CyclinA1、CDK2、PCNA、Cdc25A 及 P53 蛋白表达变化。 结果 与阴性对照组相比,miR-574-3p mimic 组 miR-574-3p 表达明显上调(P<0. 01),且在 36~ 72 h 细胞活力明显降低(P<0. 01)。 与阴性对照组相比,miR-574-3p mimic 组细胞克隆形成率显著降低(P<0. 01),同时 S 期细胞比例显著增加(P<0. 01)。 相较于阴性对照组,miR- 574-3p mimic 组 CyclinA1、CDK2、PCNA、Cdc25A 蛋白表达显著下调(P<0. 01),P53 蛋白表达明显上调(P<0. 01)。 阴性对照组与空白对照组上述各指标均无明显差异。 结论 miR-574-3p 可抑制结肠癌 HCT116 细胞增殖,这可能与下调 CyclinA1、CDK2、PCNA、Cdc25A 及上调 P53 蛋白表达,最终诱导细胞 S 期阻滞有关。

    Abstract:

    Objective Observe the effect of microRNA-574-3p (miR-574-3p) on the proliferation of colon cancer cells, and explore its potential mechanism. Methods HCT116 colon cancer cells were transfected with miR-574-3p mimics or a negative control and used miR-574-3p mimics and negative control groups, respectively. Normal HCT116 cells were used as the blank control group. RT-qPCR was used to detect expression of miR-574-3p. CCK-8 assays were used to measure cell proliferation. Colony formation assay was used to assess cell clone formation. Flow cytometry was used to detect cell cycle changes. Western blot was used to detect expression of Cyclin A1, CDK2, PCNA, Cdc25A, and p53 proteins. Results Compared with the negative control group, expression of miR-574-3p in the miR-574-3p mimics group was upregulated significantly (P<0. 01), and cell viability was significantly reduced at 36 ~ 72 h (P<0. 01). Compared with the negative control group, the colony formation rate was significantly reduced in the miR-574-3p mimics group (P<0. 01), while the proportion of S-phase cells was increased significantly (P< 0. 01). Compared with the negative control group, expression of Cyclin A1, CDK2, PCNA, and Cdc25A was significantly downregulated in the miR-574-3p mimic group (P< 0. 01) and expression of p53 protein was upregulated significantly (P<0. 01). There was no significant difference in the above indicators between negative control and blank control groups. Conclusions MiR-574-3p inhibits the proliferation of HCT116 colon cancer cells, which may be related to the downregulation of Cyclin A1, CDK2, PCNA, and Cdc25A and the upregulation of p53 protein expression, which ultimately induces cell cycle arrest in S phase.

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刘振方,孟祥彩,武 玉,刘晓飞,米永鹏. microRNA-574-3p 抑制结肠癌细胞增殖的机制研究[J].中国比较医学杂志,2021,31(10):30~35.

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  • 收稿日期:2020-12-08
  • 在线发布日期: 2021-11-29