CAJ | 학술논문

前期研究发现,过表达miR-17-92基因簇能促进鸡前脂肪细胞增殖,但作用机制尚不清楚.生物信息学分析发现,TP53INP1是miR-17-92基因簇成员miR-17-5p和miR-20a的潜在靶基因,研究采用荧光素酶报告基因技术和基因过表达技术开展该靶基因的验证.结果表明,过表达miR-17-92基因簇能显著抑制TP53INP1的3' UTR报告基因活性;而转染miR-20a抑制剂能显著提高TP53INP1的3' UTR报告基因活性.将miRNA抑制剂分别转染到DF1细胞和鸡前脂肪细胞中,采用real-time RT-PCR方法检测细胞TP53INP1基因表达变化.结果显示, miR-20a抑制剂能促进TP53INP1表达,TP53INP1是miR-20a的一个靶基因.
전기연구발현,과표체miR-17-92기인족능촉진계전지방세포증식,단작용궤제상불청초.생물신식학분석발현,TP53INP1시miR-17-92기인족성원miR-17-5p화miR-20a적잠재파기인,연구채용형광소매보고기인기술화기인과표체기술개전해파기인적험증.결과표명,과표체miR-17-92기인족능현저억제TP53INP1적3' UTR보고기인활성;이전염miR-20a억제제능현저제고TP53INP1적3' UTR보고기인활성.장miRNA억제제분별전염도DF1세포화계전지방세포중,채용real-time RT-PCR방법검측세포TP53INP1기인표체변화.결과현시, miR-20a억제제능촉진TP53INP1표체,TP53INP1시miR-20a적일개파기인.
The previous studies showed that overexpression of miR-17-92 cluster promoted the chicken preadipocyte proliferation, however, the underlying molecular mechanism remains unclear. Bioinformatics analysis found that TP53INP1 was a potential target gene of miR-17-5p and miR-20a encoded by the miR-17-92 cluster. In the present study, we verified this bioinformatics analysis result using a luciferase reporter assay and miRNA overexpression and knockdown. The results showed that miR-17-92 cluster significantly decreased TP53INP1 3'UTR luciferase reporter activity;transfection of miR-20a inhibitor significantly decreased TP53INP1 3'UTR luciferase reporter activity. Real-time RT-PCR expression analysis showed that transfection of miR-20a inhibitor increased the endogenous TP53INP1 expression in DF1cel s and immortalized chicken preadipocytes. Taken together, these data suggest that TP53INP1 is a target gene of miR-20a.