IRS1、TCONS_00056481和miR-660在猪前体脂肪细胞分化过程中的时序表达及靶向关系鉴定

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中图分类号：S828.2 文献标志码：A 文章编号：0366-6964（2025）07-3152-12

Abstract： The study aimed to explore the competitive binding and targeting relationship between TCONS_O0056481，IRS1 and miR-660. The porcine subcutaneous preadipocytes were isolated and cultured，the expression levels of TCONS_00056481，miR-660 and IRS1 in the backfat tissues of Anqing Six-end-white pigs （fat type） and Large White pigs（lean type） and their temporal expression changes during porcine preadipocytes differentiation were detected by RTqPCR，the expression levels of IRSl protein in the backfat of the two pig breeds was detected by Western blot，and the targeted binding relationship between TCONS_O0056481， IRSland miR660 were identified by dual luciferase reporter gene detection system in this study. The results showed that the expression levels of IRS1，miR-660 and TCONS_0005648l in the backfat of the two pig breeds were significantly different （ （P<0.01 ），and the expression levels of IRS1，miR660 and TCONS_Ooo56481 were significantly different during the differentiation of porcine preadipocytes， indicating that IRS1，miR-660 and TCONS_00056481 had potential regulatory roles in pig fat deposition and porcine preadipocyte differentiation. The dual luciferase activity of co-transfected TCONS_00056481 wild-type vector and miR-660 mimics was significantly lower than that of its control group （ P<0.01 ），and the dual luciferase activity of co-transfected IRS1 wild-type vector and miR-66O mimics was significantly lower than that of its control group （ （P< 0.01）；the dual luciferase activity of co-transfected TCONS_ooo56481/IRS1 mutant vector and miR-66O mimics was not significantly different from that of the control group （ P>0.05） ， indicating that miR-660 could regulate IRS1 and TCONS_O005648l through binding sites. The study preliminarily verified that lncRNA TCONS_O005648l can competitively bind to miR-660 to regulate the expression of IRS1; IRS1， TCONS_O0056481 and miR-660 can be used as candidate gene pairs of competing endogenous RNA molecules in regulating porcine lipid formation for further study.

Keywords： IRS1；miR-660；TCONS_Oo056481；dual-luciferase reporter gene assa： ∗ Corresponding author：DING Yueyun，E-mail： dingyueyun@ ahau. edu. cn

目前研究表明，人类肥胖引起的病理生理改变与糖尿病、高血压、心血管疾病、肾病以及癌症的发生密切相关[1-4]。（剩余19391字）