Dong Hui, Wang Jing, Yang Yanjuan, Zhang Xu, Yu Mengchu, Xu Guangxian, Jian Peng
Research Equipment Management Center, General Hospital of Ningxia Medical University, Yinchuan 750000, China.
Department of Pathology, General Hospital of Ningxia Medical University, Yinchuan 750000, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2018 Dec;34(12):1091-1098.
Objective To investigate the regulatory effect of miR-451a on macrophage migration inhibitory factor (MIF) and its effect on the proliferation of hepatocellular carcinoma HepG2 cells. Methods Real-time quantitative PCR was utilized to detect the expression of miR-451a and MIF mRNA in clinical liver cancer tissues. The luciferase reporter system was used to validate the regulatory relationship between miR-451a and MIF. The lentivirus overexpression vector of miR-451a was packaged to infect HepG2 cells. The expression of miR-451a and MIF mRNA was detected by real-time quantitative PCR. MIF protein level was detected by Western blotting. Cell proliferation was examined by MTT assay. Cell colony formation rate was tested by flat-plate colony formation experiment. Results The level of miR-451a was low and MIF mRNA was higher in liver cancer tissues. The luciferase reporter system showed that the intensity of the fluorescent signal was clearly weaker in MIF 3'UTR wild-type co-transfected cells with miR-451a mimics as compared with the other transfection groups. When HepG2 cells were infected with the lentivirus over-expressing miR-451a, the expression of miR-451a significantly increased; the expression of MIF significantly decreased; the cell proliferation and colony formation ability were weakened. Conclusion Overexpression of miR-451a can inhibit cell proliferation and colony formation by inhibiting MIF expression in HepG2 cells.
目的 探讨miR-451a对巨噬细胞移动抑制因子（MIF）的调控作用及其对肝癌HepG2细胞增殖的影响。方法 采用实时定量PCR检测临床肝癌组织中miR-451a和MIF mRNA的表达。利用荧光素酶报告系统验证miR-451a与MIF之间的调控关系。包装miR-451a的慢病毒过表达载体感染HepG2细胞。通过实时定量PCR检测miR-451a和MIF mRNA的表达。采用蛋白质免疫印迹法检测MIF蛋白水平。通过MTT法检测细胞增殖情况。通过平板克隆形成实验检测细胞集落形成率。结果 肝癌组织中miR-451a水平较低，MIF mRNA水平较高。荧光素酶报告系统显示，与其他转染组相比，miR-451a模拟物与MIF 3'UTR野生型共转染细胞中荧光信号强度明显较弱。当HepG2细胞感染过表达miR-451a的慢病毒时，miR-451a的表达显著增加；MIF的表达显著降低；细胞增殖和集落形成能力减弱。结论 miR-451a过表达可通过抑制HepG2细胞中MIF的表达来抑制细胞增殖和集落形成。
