Abstract
Chinese LIN28A, an RNA-binding protein, plays an important role in porcine induced pluripotent stem cells (piPSCs). However, the molecular mechanism underlying the function of LIN28A in the maintenance of pluripotency in piPSCs remains unclear. Here, we explored the function of LIN28A in piPSCs based on its overexpression and knockdown. We performed total RNA sequencing (RNA-seq) of piPSCs and detected the expression levels of relevant genes by quantitative real-time polymerase chain reaction (qRT-PCR), western blot analysis, and immunofluorescence staining. Results indicated that piPSC proliferation ability decreased following LIN28A knockdown. Furthermore, when LIN28A expression in the shLIN28A2 group was lower (by 20%) than that in the negative control knockdown group ( shNC), the pluripotency of piPSCs disappeared and they differentiated into neuroectoderm cells. Results also showed that LIN28A overexpression inhibited the expression of DUSP (dual-specificity phosphatases) family phosphatases and activated the mitogen-activated protein kinase (MAPK) signaling pathway. Thus, LIN28A appears to activate the MAPK signaling pathway to maintain the pluripotency and proliferation ability of piPSCs. Our study provides a new resource for exploring the functions of LIN28A in piPSCs. 作为RNA结合蛋白，LIN28A在猪诱导多能干细胞（piPSCs）中发挥着重要的作用。然而， LIN28A维持piPSCs多能性的分子机制尚不清楚。该文通过过表达和干扰 LIN28A来探究其在piPSCs中的作用。我们对piPSCs进行了全转录组测序（RNA-seq），并通过实时荧光定量聚合酶链式反应（qRT-PCR）、蛋白质印迹法（Western Blot）和免疫荧光染色检测相关基因的表达水平。我们的结果表明，干扰 LIN28A后，piPSCs的增殖能力明显下降。此外，当干扰组中的 LIN28A的表达量低于阴性对照组（ shNC）表达量的20%时，piPSCs会丧失多能性，并向神经外胚层细胞分化。结果还表明 LIN28A能抑制 DUSP家族磷酸酶的活性进而激活丝裂原活化蛋白激酶（MAPK）信号通路。因此， LIN28A可激活MAPK信号通路进而维持piPSCs的多能性和增殖能力。该文为探索 LIN28A在piPSCs中的作用提供了新思路。.