Nath P, Getzenberg R, Beebe D, Pallansch L, Zelenka P
Exp Cell Res. 1987 Mar;169(1):215-22. doi: 10.1016/0014-4827(87)90239-4.
Explants of the central region of lens epithelia from early chicken embryos differentiate in vitro to form lens fiber cells when cultured in the presence of chicken vitreous humor. Hybridization of a 32P-labeled v-myc viral oncogene DNA probe to RNA extracted from differentiating explants and immobilized on nitrocellulose filters indicates that levels of 2.5 kb c-myc mRNA are transiently elevated 5-10-fold in the differentiating cells. Increased levels of c-myc mRNA are observed within 30 min of the initiation of differentiation in vitro and persist for 8-9 h. Thymidine labeling of nuclei in differentiating explants indicates that entry of cells into S phase is inhibited during this period, as differentiating cells complete a final round of mitosis and withdraw from the cell cycle. Levels of c-myc mRNA are also elevated in the peripheral region of the lens epithelium, which contains cells undergoing differentiation in vivo, suggesting that the regulation of c-myc mRNA which occurs in vitro may also occur in vivo. c-myc mRNA, c-fos mRNA, and c-src mRNA showed distinct patterns of regulation associated with lens fiber formation in vivo, thus providing evidence that the regulation of c-myc mRNA is specific to this proto-oncogene. The finding that c-myc mRNA undergoes a specific, transient elevation in differentiating lens cells as they withdraw from the cell cycle contrasts with a large body of evidence linking enhanced c-myc expression with increased cell proliferation.
来自早期鸡胚晶状体上皮中央区域的外植体,在鸡玻璃体液存在的情况下进行体外培养时会分化形成晶状体纤维细胞。用32P标记的v-myc病毒癌基因DNA探针与从正在分化的外植体中提取并固定在硝酸纤维素滤膜上的RNA进行杂交,结果表明,在分化细胞中,2.5 kb的c-myc mRNA水平会短暂升高5至10倍。在体外分化开始后30分钟内即可观察到c-myc mRNA水平升高,并持续8至9小时。对正在分化的外植体细胞核进行胸苷标记表明,在此期间细胞进入S期受到抑制,因为分化细胞完成最后一轮有丝分裂并退出细胞周期。在晶状体上皮的周边区域,c-myc mRNA水平也会升高,该区域含有体内正在分化的细胞,这表明体外发生的c-myc mRNA调控在体内可能也会发生。c-myc mRNA、c-fos mRNA和c-src mRNA在体内与晶状体纤维形成相关的调控模式各不相同,因此提供了证据表明c-myc mRNA的调控对该原癌基因具有特异性。c-myc mRNA在分化的晶状体细胞退出细胞周期时会经历特定的、短暂的升高,这一发现与大量将增强的c-myc表达与细胞增殖增加联系起来的证据形成了对比。