miR-451a通过下调巨噬细胞移动抑制因子抑制胶质瘤增殖的研究

miR-451a suppresses proliferation of gliomay by down-regulating macrophage migration inhibitory factors

目的研究癌基因巨噬细胞移动抑制因子(MIF)受miR-451a负调控构成的调控网络在胶质瘤发生发展中的作用。方法qRT-PCR检测miR-451a在27例胶质瘤患者组织样本和细胞系中的表达;采用Western blot及qRT-PCR检测胶质瘤组织中MIF的表达变化;双荧光素酶报告基因检验靶向调控关系;Western blot检测miR-451a-MIF轴的调控关系;Transwell、细胞划痕和TUNEL实验分别检测miR-451a-MIF轴对细胞侵袭、迁移和凋亡的影响;构建胶质瘤裸鼠皮下移植瘤模型,采用多组增殖细胞核抗原(PCNA)实验检测miR-451a-MIF轴对体内细胞增殖的影响。结果miR-451a在胶质瘤组织和细胞系中均低表达,MIF在胶质瘤组织和细胞系中均上调。MIF受miR-451a负调控。体外细胞实验中miR-451a抑制U87细胞侵袭、迁移及促进凋亡,而MIF逆转miR-451a的抑制作用。体内实验中miR-451a抑制体内细胞增殖,同样MIF抑制miR-451a减少体内细胞增殖的能力。结论miR-451a可能通过负调控MIF的蛋白表达,参与胶质瘤的发生发展。

Objective To study the effect of the regulatory network composed of oncogene macrophage migration inhibitory factor(MIF)by the negative regulation of miR-451a in the occurrence and development of glioma.Methods qRT-PCR was used to detect the miR-451a expression in tissue specimens from 27 glioma patients and cell lines.Western blot was used to detect MIF expression in glioma tumor tissues.The dual-luciferase reporter gene was used to detect the target regulatory relationship.Western blot was used to detect the regulatory relationship of the miR-451a-MIF axis.Transwell,scratch-wound healing and TUNEL experiments were used to detect the effects of the miR-451a-MIF axis on invasion,migration and apoptosis,respectively.The nude mouse subcutaneous xenograft model of glioma was established,and a proliferating cell nuclear antigen(PCNA)assay was used to detect the effects of the miR-451a-MIF axis on cell proliferation in vivo.Results miR-451a was lowly expressed in glioma tissues and cell lines.MIF was up-regulated in glioma tissue and cell lines.MIF was negatively regulated by miR-451a.In the in vitro cell experiment,miR-451a inhibited the invasion and migration of U87 cells and promoted their apoptosis,while MIF reversed the inhibitory effects of miR-451a.In the in vivo experiment,miR-451a inhibited the in vivo cell proliferation,and MIF similarly inhibited miR-451a to decrease the in vivo cell proliferation.Conclusion miR-451a participates in the occurrence and development of glioma possibly by negatively regulating MIF protein expression.