miR-351-3p promotes rat amniotic fluid-derived mesenchymal stromal cell proliferation via targeting the coding sequence of Abca4,STEM CELLS

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miR-351-3p promotes rat amniotic fluid-derived mesenchymal stromal cell proliferation via targeting the coding sequence of Abca4
STEM CELLS ( IF 4.0 ) Pub Date : 2021-05-10 , DOI: 10.1002/stem.3392
Jieting Huang _{1,

2,

3} , Qi Zhao ₄ , Xiaowei Wei ₃ , Wei Ma ₃ , Wenting Luo ₃ , Hui Gu ₃ , Dan Liu ₃ , Yiwen He ₃ , Tianchu Huang ₃ , Yusi Liu ₅ , Chenfei Wang ₃ , Zhengwei Yuan ₃

Affiliation

Amniotic fluid-derived mesenchymal stromal cells (AFMSCs) present different features, depending on the isolation timing and culture conditions. The lack of uniform experimental standards hinders the comparison of results from different studies on AFMSCs. Moreover, understanding the molecular mechanisms that underlie the features of AFMSCs isolated at different embryonic developmental stages might allow the obtention of more viable and highly proliferative AFMSCs through genetic modification. We isolated AFMSCs from pregnant rats at embryonic day (E)12, E15, E18, and E21 and compared their cell proliferation capacity and transcriptome. The cell counting kit-8 assay and RNA sequencing revealed that E12 and E15 AFMSCs showed different characteristics from E18 and E21 AFMSCs. Therefore, AFMSCs were divided into two groups: early (E12 and E15) and late (E18 and E21) pregnancy-stage groups. Next, we screened the gene/microRNA pair Abca4/miR-351-3p that was related to cell proliferation. Abca4 knockdown/overexpression suggested that this gene represses the proliferation of AFMSCs, which is a newly discovered function of this gene. Finally, dual luciferase reporter gene assays confirmed that miR-351-3p targeted the coding sequence of Abca4 and regulated AFMSC proliferation. miR-351-3p promotes AFMSC proliferation via targeting the coding sequence of Abca4. Our findings provide a molecular foundation for further research for obtaining AFMSCs with a higher proliferation capacity.

中文翻译：

miR-351-3p通过靶向Abca4编码序列促进大鼠羊水间充质基质细胞增殖

羊水来源的间充质基质细胞 (AFMSCs) 呈现出不同的特征，这取决于分离时间和培养条件。缺乏统一的实验标准阻碍了对 AFMSCs 不同研究结果的比较。此外，了解在不同胚胎发育阶段分离的 AFMSCs 特征的分子机制可能允许通过基因改造获得更可行和高度增殖的 AFMSCs。我们在胚胎期 (E)12、E15、E18 和 E21 从怀孕大鼠中分离了 AFMSCs，并比较了它们的细胞增殖能力和转录组。细胞计数 kit-8 测定和 RNA 测序显示 E12 和 E15 AFMSCs 显示出与 E18 和 E21 AFMSCs 不同的特征。因此，AFMSCs 分为两组：早期（E12 和 E15）和晚期（E18 和 E21）妊娠期组。接下来，我们筛选了基因/microRNA 对Abca4 /miR-351-3p 与细胞增殖有关。Abca4敲低/过表达表明该基因抑制 AFMSCs 的增殖，这是该基因新发现的功能。最后，双荧光素酶报告基因检测证实 miR-351-3p 靶向Abca4的编码序列并调节 AFMSC 增殖。miR-351-3p 通过靶向Abca4的编码序列促进 AFMSC 增殖。我们的研究结果为进一步研究获得具有更高增殖能力的AFMSCs提供了分子基础。

更新日期：2021-05-10

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