miR-140 inhibits porcine fetal fibroblasts proliferation by directly targeting type 1 insulin-like growth factor receptor and indirectly inhibiting type 1 insulin-like growth factor receptor expression via SRY-box 4
miR-140 inhibits porcine fetal fibroblasts proliferation by directly targeting type 1 insulin-like growth factor receptor and indirectly inhibiting type 1 insulin-like growth factor receptor expression via SRY-box 4
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miR-140通过直接靶向1型胰岛素样生长因子受体并通过SRY-box 4间接抑制1型胰岛素样生长因子受体表达来抑制猪胎儿成纤维细胞增殖
DOI：
10.5713/ajas.19.0438
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发表时间：
2019-11
期刊：
影响因子：
--
通讯作者：
Liu Songcai
中科院分区：
文献类型：
--
作者：
Geng Hongwei;Hao Linlin;Cheng Yunyun;Wang Chunli;Wei Wenzhen;Yang Rui;Li Haoyang;Zhang Ying;Liu Songcai
目的研究miR-140对猪胎儿成纤维细胞（pff）增殖的影响，并鉴定miR-140在pff中的靶基因。方法采用生物信息学软件对miR-140的靶基因进行预测和验证。采用定量聚合酶链反应和western blot检测pff中miR-140与其靶基因的关系。采用双荧光素酶报告基因检测来评估miR-140、1型胰岛素样生长因子受体（IGF1R）和SRY-box 4 （SOX4）之间的相互作用。当转染miR-140模拟物或抑制剂时，通过CCK-8测量miR-140对pff增殖的影响。采用染色质免疫沉淀法（ChIP）检测IGF1R启动子结合的转录因子SOX4。结果miR-140直接靶向IGF1R，抑制pff的增殖。同时，miR-140靶向与猪IGF1R启动子结合的转录因子SOX4，间接抑制IGF1R的表达。此外，miR-140抑制剂促进PFFs增殖，这一作用被SOX4或IGF1R敲低所消除。结论miR-140通过直接靶向IGF1R，并通过SOX4间接抑制IGF1R的表达，抑制PFFs增殖，在猪胎儿发育中发挥重要作用。
Objective This study aimed to elucidate the effect of miR-140 on the proliferation of porcine fetal fibroblasts (PFFs) and identify the target genes of miR-140 in PFFs. Methods In this study, bioinformatics software was used to predict and verify target genes of miR-140. Quantitative polymerase chain reaction and western blot were used to detect the relationship between miR-140 and its target genes in PFFs. Dual luciferase reporter gene assays were performed to assess the interactions among miR-140, type 1 insulin-like growth factor receptor (IGF1R), and SRY-box 4 (SOX4). The effect of miR-140 on the proliferation of PFFs was measured by CCK-8 when PFFs were transfected with a miR-140 mimic or inhibitor. The transcription factor SOX4 binding to promoter of IGF1R was detected by chromatin immunoprecipitation assay (ChIP). Results miR-140 directly targeted IGF1R and inhibited proliferation of PFFs. Meanwhile, miR-140 targeted transcription factor SOX4 that binds to promoter of porcine IGF1R to indirectly inhibit the expression of IGF1R. In addition, miR-140 inhibitor promoted PFFs proliferation, which is abrogated by SOX4 or IGF1R knockdown. Conclusion miR-140 inhibited PFFs proliferation by directly targeting IGF1R and indirectly inhibiting IGF1R expression via SOX4, which play an important role in the development of porcine fetal.
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