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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 explore whether miR-23b-3p targets hepatic nuclear factor-4 and down-regulates apo (a) expression, providing a novel approach to reduce the level of lipoprotein (a). Methods Target gene analysis of miR-23b-3p and transcription factor hepatic nuclear factor-4 (HNF4), which regulates LPA gene, was performed using Targetscan online tool. The luciferase reporter system was used in miR109 cells to screen miR-23b-3p LPA gene transcription factor HNF4 target gene validation experiments; RT-PCR Hep G2 cells apo (a) mRNA expression levels. Results Targetscan bioinformatics analysis tools showed that HNF4G could serve as the target gene of miR-23b-3p. The fluorescence intensity of miR-23b-3p-treated cells transfected with luciferase reporter system was significantly lower than that of the control group. MiR-23b-3p HNF4G can be used as the target gene of miR-23b-3p. Transfection of miR-23b-3p inhibited apo (a) mRNA expression in Hep G2 cells. Conclusion miR-23b-3p takes HNF4 as its target gene and down-regulates apo (a) expression in Hep G2 cells.