Pipkin J L, Hinson W G, Anson J F, Lyn-Cook L E, Burns E R, Casciano D A
Food and Drug Administration, National Center for Toxicological Research, Jefferson, Arkansas 72079.
Appl Theor Electrophor. 1991;2(1):17-29.
Exposure of HL-60 cells to 2 microM retinoic acid, twice the dose necessary for differentiation, initiated protein synthesis within 2 h exposure in the nuclear matrix proteins and phosphorylation of a subfraction of these proteins, designated the phenol-soluble nuclear proteins. These processes were examined by fluorography of two-dimensional polyacrylamide gels. Three cell-cycle related stress proteins (22, 70c, 70x Mr x 10(-3)), were seen in the nuclear matrix fraction that were synthesized early and disappeared rapidly following dosing with retinoic acid. In control cells, protein 120 was also cell-cycle related and showed modest synthesis in nuclear matrix and strong phosphorylation in phenol-soluble fraction. Within 5 h after dosing, p120 exhibited alteration in phosphorylation as evidenced by mapping of [32P]-labeled peptides. Two nuclear matrix proteins, p52 and p55, incorporated [3H] retinoic acid rapidly, were cell-cycle-related, and disappeared within 12 h of dosing. Progressive increases in the labeling of the nuclear matrix and phenol-soluble nuclear proteins with increasing retinoic acid exposure was apparent in G2-phase at 96 h time-after-dosing.
将HL-60细胞暴露于2微摩尔视黄酸（该剂量是诱导分化所需剂量的两倍）下，在暴露2小时内，核基质蛋白开始合成，并且这些蛋白的一个亚组分（称为酚溶性核蛋白）发生磷酸化。通过二维聚丙烯酰胺凝胶的荧光自显影来检测这些过程。在核基质组分中观察到三种与细胞周期相关的应激蛋白（分子量分别为22、70c、70x×10⁻³），它们在给药视黄酸后早期合成并迅速消失。在对照细胞中，蛋白120也与细胞周期相关，在核基质中合成适度，在酚溶性组分中磷酸化强烈。给药后5小时内，通过对[³²P]标记肽的图谱分析表明，p120的磷酸化发生了改变。两种核基质蛋白p52和p55迅速掺入[³H]视黄酸，与细胞周期相关，并在给药后12小时内消失。在给药后96小时的G2期，随着视黄酸暴露增加，核基质和酚溶性核蛋白的标记逐渐增加。
