摘要
目的观察微小RNA-574-3p(micro RNA-574-3p,mi R-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mi R-574-3p mimic和阴性对照分别作为mi R-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mi R-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Cyclin A1、CDK2、PCNA、Cdc25A及P53蛋白表达变化。结果与阴性对照组相比,mi R-574-3p mimic组mi R-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,mi R-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,mi R-574-3p mimic组Cyclin A1、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mi R-574-3p可抑制结肠癌HCT116细胞增殖,这可能与下调Cyclin A1、CDK2、PCNA、Cdc25A及上调P53蛋白表达,最终诱导细胞S期阻滞有关。
Objective Observe the effect of micro RNA-574-3 p(mi R-574-3 p)on the proliferation of colon cancer cells,and explore its potential mechanism.Methods HCT116 colon cancer cells were transfected with mi R-574-3 p mimics or a negative control and used mi R-574-3 p mimics and negative control groups,respectively.Normal HCT116 cells were used as the blank control group.RT-qPCR was used to detect expression of mi R-574-3 p.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,Cdc25 A,and p53 proteins.Results Compared with the negative control group,expression of mi R-574-3 p in the mi R-574-3 p 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 mi R-574-3 p 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 Cdc25 A was significantly downregulated in the mi R-574-3 p 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-574-3 p inhibits the proliferation of HCT116 colon cancer cells,which may be related to the downregulation of Cyclin A1,CDK2,PCNA,and Cdc25 A and the upregulation of p53 protein expression,which ultimately induces cell cycle arrest in S phase.
作者
刘振方
孟祥彩
武玉
刘晓飞
米永鹏
LIU Zhenfang;MENG Xiangcai;WU Yu;LIU Xiaofei;MI Yongpeng(Department of General Surgery,Shijiazhuang Hospital of Traditional Chinese Medicine,Shijiazhuang 050051,China;Department of General Surgery,First Hospital of Qinhuangdao,Qinhuangdao 066000)
出处
《中国比较医学杂志》
CAS
北大核心
2021年第10期30-35,共6页
Chinese Journal of Comparative Medicine
基金
河北省2020年度医学科学研究课题计划(20201428)。
