School of Life Sciences, Technical University of Munich, 85354 Freising, Germany.
Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Taian 271018, China.
Genes (Basel). 2022 Jun 3;13(6):1013. doi: 10.3390/genes13061013.
MicroRNA-143-3p (miR-143-3p) is one of the miRNAs involved in the growth of goat mammary epithelial cells (GMECs). In this study, Illumina/Solexa sequencing was performed to establish the lncRNA database in Laoshan dairy goats. Using the lncRNA database, long noncoding RNAs (lncRNAs) regulated by miR-143-3p were screened. In total, 4899 lncRNAs were identified, with 173 lncRNAs being differentially expressed in all three replicates. The target genes of the differentially expressed lncRNAs were annotated in GO terms and KEGG pathways. Among the differentially expressed lncRNAs, lncRNA LOC102188416 was predicted to sponge miR-143-3p and share as a common target gene with miR-143-3p, which was validated by dual luciferase reporter assay system and qRT-PCR. The miR-143-3p mimic significantly lowered the relative luciferase activity of psiCHECK2-LOC102188416 wildtype vector but not mutated vector, suggesting that lncRNA LOC102188416 might be a sponge of miR-143-3p, which was verified by the promotion role of lncRNA LOC102188416 siRNA (siR-LOC102188416) in the expression of miR-143-3p. It was shown that the expression of was downregulated by either miR-143-3p mimic or siR-LOC102188416, indicating that miR-143-3p and lncRNA LOC102188416 had a coregulatory effect on expression. The co-transfection of miR-143-3p inhibitor with siR-LOC102188416 reversed the decrease of expression regulated by siR-LOC102188416 alone, strengthening the existence of lncRNA LOC102188416/miR-143-3p/ axis in GMECs of Laoshan dairy goats.
miR-143-3p（miR-143-3p）是参与山羊乳腺上皮细胞（GMECs）生长的miRNAs 之一。在这项研究中，使用 Illumina/Solexa 测序在崂山奶山羊中建立了 lncRNA 数据库。利用 lncRNA 数据库，筛选受 miR-143-3p 调控的长非编码 RNA（lncRNA）。总共鉴定出 4899 个 lncRNA，其中 173 个 lncRNA 在所有三个重复中均有差异表达。差异表达 lncRNA 的靶基因被注释在 GO 术语和 KEGG 途径中。在差异表达的 lncRNA 中，lncRNA LOC102188416 被预测为 miR-143-3p 的海绵，并与 miR-143-3p 共享 作为共同靶基因，这通过双荧光素酶报告基因检测系统和 qRT-PCR 得到了验证。miR-143-3p 模拟物显著降低 psiCHECK2-LOC102188416 野生型载体的相对荧光素酶活性，但不降低突变型载体的相对荧光素酶活性，表明 lncRNA LOC102188416 可能是 miR-143-3p 的海绵，这通过 lncRNA LOC102188416 siRNA（siR-LOC102188416）在 miR-143-3p 表达中的促进作用得到了验证。结果表明，miR-143-3p 模拟物或 siR-LOC102188416 均可下调 的表达，表明 miR-143-3p 和 lncRNA LOC102188416 对 的表达具有协同调控作用。miR-143-3p 抑制剂与 siR-LOC102188416 共转染可逆转 siR-LOC102188416 单独调节的 的表达降低，这增强了 lncRNA LOC102188416/miR-143-3p 轴在崂山奶山羊 GMECs 中的存在。
