Production and purification of recombinant buffalo leukemia inhibitory factor (rBuLIF) from stably transfected COS-1 cells-猫眼课题宝

从稳定转染的 COS-1 细胞中生产和纯化重组水牛白血病抑制因子 (rBuLIF)

DOI：

10.1096/fj.99-0810com

发表时间：

2015

期刊：

The FASEB Journal

影响因子：

通讯作者：

中科院分区：

文献类型：

作者：

G. Kaur;N. Das;S. Kumar

小鼠胚胎干细胞在白血病抑制因子（LIF）存在下体外生长时仍具有多能性。LIF退出导致ES细胞进行性分化。在这里，我们表明，在这个分化过程中，部分细胞发生凋亡伴随着激活的p38 MAP激酶。为了深入了解ES细胞中由LIF介导的事件，通过使用半定量RT-PCR测定在不存在或存在该细胞因子的情况下分析潜在候选基因的表达。我们专注于早期反应基因和一种新型的细胞因子抑制因子（Socs蛋白），其中一些具有抗凋亡特性。我们发现，c‐Fos、c‐Jun和JunB的表达在LIF处理后被诱导，而JunD、酪氨酸磷酸酶ESP和LIF受体的组分的表达不受影响。Socs-3的表达，但不是Socs-1或Socs-2，在LIF的存在下被刺激。最后，Socs-1和Socs-3的不受控制的过表达导致LIF依赖性转录的抑制和细胞活力的严重降低，表明良好平衡的Socs蛋白含量的干扰对细胞存活具有不利影响。Reinhardt，B.，Kedinger，C.，Boeuf，H.细胞因子信号传导抑制因子（Socs）在白血病抑制因子（LIF）依赖性胚胎干细胞存活中的作用.FASEB J.14，1577-1584（2000）

Mouse embryonic stem (ES) cells remain pluripotentin vitrowhen grown in the presence of leukemia inhibitory factor (LIF). LIF withdrawal results in progressive ES cell differentiation. Here we show that during this differentiation process, part of the cells undergo apoptosis concomitant with an activation of the p38 MAP kinase. To gain insight into events mediated by LIF in ES cells, the expression of potential candidate genes was analyzed in the absence or presence of this cytokine by using a semiquantitative RT‐PCR assay. We focused on early response genes and on a new type of cytokine repressors (the Socs proteins), some of which exhibit anti‐apoptotic properties. We found that expression of c‐Fos, c‐Jun, and JunB was induced upon LIF treatment whereas that of JunD, the tyrosine phosphatase ESP, and the components of the LIF receptor remained unaffected. Expression of Socs‐3, but not Socs‐1 or Socs‐2, was stimulated in the presence of LIF. Finally, uncontrolled overexpression of Socs‐1 and Socs‐3 led to repression of LIF‐dependent transcription and severely reduced cell viability, suggesting that the disturbance of a well balanced Socs protein content has adverse effects on cell survival.–Duval, D., Reinhardt, B., Kedinger, C., Boeuf, H. Role of suppressors of cytokine signaling (Socs) in leukemia inhibitory factor (LIF) ‐dependent embryonic stem cell survival.FASEB J.14, 1577–1584 (2000)