LncRNA-599547 contributes the inductive property of dermal papilla cells in cashmere goat through miR-15b-5p/Wnt10b axis

LncRNA-599547 contributes the inductive property of dermal papilla cells in cashmere goat through miR-15b-5p/Wnt10b axis
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LncRNA-599547 通过 miR-15b-5p/Wnt10b 轴贡献绒山羊真皮乳头细胞的诱导特性

DOI:
10.1080/10495398.2020.1806860
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发表时间:
2020-08
期刊:
Animal Biotechnology, 2020,DOI: 10.1080/10495398.2020.1806860
影响因子:
--
通讯作者:
Bai Wen L.
Bai Wen L.
中科院分区:
其他
文献类型:
--
作者:
Yin Rong H.;Zhao Su J.;Wang Ze Y.;Zhu Yu B.;Yin Rong L.;Bai Man;Fan Yi X.;Wang Wei;Bai Wen L.

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摘要在绒山羊次生毛囊(SHF)中鉴定到lncRNA-599547(长度为619 nt),但其在调节真皮乳头细胞(DPCs)诱导特性中的功能作用尚不清楚。我们发现lncRNA-599547在绒山羊SHF生长初期的真皮乳头中的表达明显高于其在休止期的表达。lncRNA-599547过表达导致DPCs中ALP和LEF1的表达显著升高(p < 0.05),而siLncRNA-1介导的lncRNA-599547沉默则显著下调DPCs中ALP和LEF1的表达(p < 0.05)。基于生物素标记RNA下拉实验,我们发现lncRNA-599547在DPCs中直接与chi-miR-15b-5p相互作用。基于lncRNA-599547的过表达和沉默分析,我们的研究结果表明,lncRNA-599547促进Wnt10b在DPCs中的表达,但不调节其启动子甲基化水平。利用双荧光素酶报告基因检测中含有chi-miR-15b-5p预测结合位点的山羊Wnt10b mRNA-3'UTR片段,我们发现lncRNA-599547在chi-miR-15b-5p介导的转录后水平上调节Wnt10b的表达。综上所述,我们的研究结果表明,lncRNA-599547诱导miR-15b-5p正向调节Wnt10基因的表达,从而促进了绒山羊DPCs的诱导特性。
Abstract The lncRNA-599547 (619-nt in length) is identified in secondary hair follicle (SHF) of cashmere goat, but its functional roles in regulating the inductive property of dermal papilla cells (DPCs) remains unknown. We found that lncRNA-599547 had significantly higher expression in dermal papilla of cashmere goat SHF at anagen than its counterpart at telogen. The overexpression of lncRNA-599547 led to a significant increase of ALP and LEF1 expression in DPCs (p < 0.05), whereas, the siLncRNA-1 mediated silencing of lncRNA-599547 significantly down-regulated the expression of ALP and LEF1 in DPCs (p < 0.05). Based on biotin-labeled RNA pull-down assay, we found that lncRNA-599547 directly interacted with chi-miR-15b-5p in DPCs. Based on both overexpression and silencing analysis of lncRNA-599547, our results indicate that lncRNA-599547 promotes the expression of Wnt10b in DPCs but without modulating its promoter methylation level. Using the mRNA-3′UTR fragments of goat Wnt10b containing the predicted binding sites of chi-miR-15b-5p in Dual-luciferase Reporter Assays, we show that lncRNA-599547 modulates the expression of Wnt10b at the chi-miR-15b-5p mediated post-transcriptional level. Taken together, our results indicate that lncRNA-599547 sponges miR-15b-5p to positively regulate the expression of Wnt10 gene, and thereby contributes the inductive property of DPCs in cashmere goat.
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