miR⁃148a⁃3p靶向DLL4基因影响婴幼儿血管瘤内皮细胞增殖和凋亡

miR⁃148a⁃3p inhibits proliferation and promotes apoptosis of infantile hemangioma endothelial cells by targeting DLL4

目的探讨miR⁃148a⁃3p靶向Delta样配体4(DLL4)对婴幼儿血管瘤内皮细胞增殖和凋亡的影响。方法实时荧光定量PCR(RT⁃qPCR)、蛋白质印记(Western blot)检测miR⁃148a⁃3p、DLL4在血管瘤组织、正常皮肤组织中的表达。将血管瘤内皮细胞分为miR⁃NC、miR⁃148a⁃3p、si⁃NC、si⁃DLL4、miR⁃148a⁃3p+pcDNA、miR⁃148a⁃3p+pcDNA⁃DLL4组。采用四甲基偶氮唑盐(MTT)法、流式细胞术检测细胞活力和凋亡。双荧光素酶报告实验和Western blot确定miR⁃148a⁃3p和DLL4的调控关系。结果与正常皮肤组织比较,血管瘤组织中miR⁃148a⁃3p表达降低,DLL4表达增加,差异有统计学意义(P<0.05)。过表达miR⁃148a⁃3p或抑制DLL4后,血管瘤内皮细胞活力降低,凋亡率增加,差异有统计学意义(P<0.05)。与过表达miR⁃148a⁃3p比较,过表达DLL4可逆转miR⁃148a⁃3p对血管瘤内皮细胞增殖、凋亡的影响,差异有统计学意义(P<0.05)。miR⁃148a⁃3p靶向负调控DLL4表达。结论本研究表明miR⁃148a⁃3p通过靶向DLL4可抑制血管瘤内皮细胞增殖,促进细胞凋亡,是婴幼儿血管瘤的潜在治疗靶点。

Objective To investigate the effects of miR⁃148a⁃3p on the proliferation and apoptosis of infantile hemangioma endothelial cells by targeting Delta⁃like ligand 4(DLL4).Methods The expressions of miR⁃148a⁃3p and DLL4 in hemangiomas and normal skin were detected by real⁃time quantitative PCR(RT⁃qPCR)and Western blot.Hemangioma endothelial cells were divided into miR⁃NC,miR⁃148a⁃3p,si⁃NC,si⁃DLL4,miR⁃148a⁃3p+pcDNA,miR⁃148a⁃3p+pcDNA⁃DLL4 groups.Methyl thiazolyl tetrazolium(MTT)method and flow cytometry were utilized to detect cell viability and apoptosis.Results Compared with the normal skin tissue,the expression of miR⁃148a⁃3p was decreased in hemangiomas,while the expression of DLL4 was increased,the difference was statistically significant(P<0.05).Over⁃expression of miR⁃148a⁃and down⁃regulation of DLL4,hemangioma endothelial cell viability was decreased,and the apoptosis rate was in⁃creased,the difference was statistically significant(P<0.05).Compared with the over⁃expression of miR⁃148a⁃3p,over⁃expression of DLL4 reversed the effects of miR⁃148a⁃3p on hemangioma endothelial cell proliferation and apoptosis,the difference was statistically significant(P<0.05).miR⁃148a⁃3p negatively regulates DLL4 expression.Conclusion miR⁃148a⁃3p inhibits the proliferation and promotes apoptosis of hemangioma endo⁃thelial cells by targeting DLL4,suggesting a potential therapeutic target for infantile hemangiomas.