miR-23b-3p对肝核因子4的靶基因及效用分析

Target gene and effect analysis of miR-23b-3p on hepatocyte nuclear factor 4

目的探索miR-23b-3p是否以肝核因子4作为靶基因并下调apo(a)表达,为降高脂蛋白(a)血症提供新的思路。方法用Targetscan在线工具对miR-23b-3p与调控LPA基因的转录因子肝核因子4(HNF4)进行靶基因分析;在JM109细胞中使用荧光素酶报告系统对miR-23b-3p与调控筛选调控LPA基因的转录因子HNF4进行靶基因进行验证实验;用RT-PCR检测Hep G2细胞apo(a)mRNA表达水平。结果 Targetscan生物信息学分析工具表明HNF4G可作为miR-23b-3p的靶基因,荧光素酶报告系统转染miR-23b-3p处理组细胞裂解后荧光强度显著低于对照组,miR-23b-3p的抑制剂可抑制其作用,验证了HNF4G可作为miR-23b-3p的靶基因。转染miR-23b-3p可抑制Hep G2细胞apo(a)mRNA的表达。结论 miR-23b-3p以HNF4为其靶基因,并下调Hep G2细胞apo(a)的表达。

Objective To investigate whether or not hepatocyte nuclear factor 4（ HNF4） is the target gene of miR-23b-3p,and down-regulating effect of miR-23b-3p on apolipoprotein（ a） [apo（ a） ] expression,providing a new idea for reducing high concentration lipoprotein（ a）. Methods Target genes in miR-23b-3p and hepatocyte nuclear factor 4 were analyzed by Targetgene,an online bioinformatics tools,and tested by dual luciferase reporter assays in JM109 cell,RT-PCR was used to measure the expression level of apo（ a） in Hep G2 cell. Results Targetgene scanning result indicated that HNF4 mRNA could be target gene of a miR-23b-3p;,compared with the control group,the fluorescence density of the group with miR-23b-3p transfection in dual luciferase reporter assays system,decreased significantly; miR-23b-3p inhibitor could inhibit the role of miR-23b-3p,which verified that HNF4 is a target gene of miR-23b-3p; and apo（ a） mRNA expression decreased after miR-23b-3p transfection in Hep G2 cell. Conclusion HNF4 is a target gene of miR-23b-3pand it can decrease apo（ a） mRNA expression.