Pan H, Griep A E
Department of Anatomy, University of Wisconsin Medical School, Madison 53706.
Genes Dev. 1994 Jun 1;8(11):1285-99. doi: 10.1101/gad.8.11.1285.
Tumor suppressor proteins are believed to play a role in regulating cell cycle control during mammalian development. The E6 and E7 oncoproteins from human papillomavirus type 16 are known to affect cell growth control, at least in part, through their inactivation of cellular tumor suppressor gene products, p53 and Rb, respectively. Therefore, these viral proteins can serve as trans-dominant repressors of tumor suppressor gene function. To study the potential role of p53 and Rb in murine lens morphogenesis, we generated transgenic mice in which the expression of E6 or E7 was directed to the developing lens. Transgenic mice expressing E7 exhibited microphthalmia and cataracts, whereas transgenic mice expressing E6 exhibited cataracts without noticeable microphthalmia. Microscopic analysis of the lenses from neonatal and adult E7 transgenic mice revealed inhibition of lens fiber cell differentiation, induction of cell proliferation in spatially inappropriate regions of the lens, and apoptosis. Transgenic mice expressing a mutant E7 that is defective in Rb/p107 binding exhibited normal eyes, suggesting that the activity of Rb and/or Rb-like proteins is required for the perturbation of lens development and induction of apoptosis in E7 mice. Microscopic analysis of lenses from E6 neonatal and adult transgenic mice indicated the presence of nuclei in elongated fiber cells, suggesting that E6 inhibits lens fiber cell denucleation. Furthermore, expression of E6 inhibited the apoptotic-like DNA degradation observed in the lenses of nontransgenic 15.5-day embryos. In lenses from neonatal E6 x E7 double transgenic mice, the level of apoptosis was reduced compared with that seen in lenses from neonatal E7 mice. In adults E6 x E7 double transgenic mice, lens tumors developed, whereas in E6 or E7 only transgenic mice, tumors did not. Taken together, these results point to specific roles in lens morphogenesis for Rb and p53 and to the necessity of these tumor suppressor gene products in regulating exit from the normal cell division cycle in differentiating lens fiber cells.
肿瘤抑制蛋白被认为在哺乳动物发育过程中调节细胞周期控制方面发挥作用。已知来自人乳头瘤病毒16型的E6和E7癌蛋白至少部分地通过分别使细胞肿瘤抑制基因产物p53和Rb失活来影响细胞生长控制。因此,这些病毒蛋白可作为肿瘤抑制基因功能的反式显性抑制剂。为了研究p53和Rb在小鼠晶状体形态发生中的潜在作用,我们构建了将E6或E7的表达定向到发育中的晶状体的转基因小鼠。表达E7的转基因小鼠表现出小眼症和白内障,而表达E6的转基因小鼠表现出白内障但无明显的小眼症。对新生和成年E7转基因小鼠晶状体的显微镜分析显示晶状体纤维细胞分化受到抑制,在晶状体空间不适当区域细胞增殖增加,以及细胞凋亡。表达在Rb/p107结合方面有缺陷的突变型E7的转基因小鼠眼睛正常,这表明Rb和/或Rb样蛋白的活性是E7小鼠晶状体发育紊乱和细胞凋亡诱导所必需的。对E6新生和成年转基因小鼠晶状体的显微镜分析表明,细长纤维细胞中存在细胞核,这表明E6抑制晶状体纤维细胞去核。此外,E6的表达抑制了在非转基因15.5天胚胎晶状体中观察到的凋亡样DNA降解。在新生E6×E7双转基因小鼠的晶状体中,与新生E7小鼠晶状体相比,细胞凋亡水平降低。在成年E6×E7双转基因小鼠中,晶状体肿瘤发生,而在仅表达E6或E7的转基因小鼠中,未发生肿瘤。综上所述,这些结果表明Rb和p53在晶状体形态发生中具有特定作用,并且这些肿瘤抑制基因产物对于调节分化的晶状体纤维细胞从正常细胞分裂周期退出是必需的。