Missero C, Serra C, Stenn K, Dotto G P
Department of Pathology, Yale University, New Haven, Connecticut 06510.
J Cell Biol. 1993 Jun;121(5):1109-20. doi: 10.1083/jcb.121.5.1109.
In cultured cells, mutants of the Adenovirus E1a oncoprotein which bind to a reduced set of cellular proteins, including p105-Rb, p107, and p60-cyclin A, are transformation defective but can still interfere with exogenous growth inhibitory and differentiating signals, such as those triggered by TGF-beta. We have tested the ability of one such mutant, NTdl646, to interfere with keratinocyte growth and differentiation in vivo, in the skin of transgenic mice. Keratinocyte-specific expression of the transgene was achieved by using a keratin 5 promoter. Two independent lines of transgenic mice were obtained which expressed E1a specifically in their skin and exhibited an aberrant hair coat phenotype with striking regional variations. Affected hair shafts were short and crooked and hair follicles exhibited a dystrophic or absent inner root sheath. Interfollicular epidermis was normal, but its hyperplastic response to acute treatment with TPA (12-O-tetradecanoylphorbol-13-acetate) was significantly reduced. Primary keratinocytes derived from these animals were partially resistant to the effects of TPA and TGF-beta. The rate of spontaneous or chemically induced skin tumors in the transgenic mice was not increased. Thus, expression of a transgene which interferes with known negative growth regulatory proteins causes profound disturbances of keratinocyte maturation into a highly organized structure such as the hair follicle but does not lead to increased and/or neoplastic proliferation.
在培养细胞中,腺病毒E1a癌蛋白的突变体与一组减少的细胞蛋白结合,包括p105-Rb、p107和p60-细胞周期蛋白A,这些突变体具有转化缺陷,但仍能干扰外源性生长抑制和分化信号,如由TGF-β触发的信号。我们已经测试了其中一种突变体NTdl646在转基因小鼠皮肤中干扰角质形成细胞生长和分化的能力。通过使用角蛋白5启动子实现转基因在角质形成细胞中的特异性表达。获得了两个独立的转基因小鼠品系,它们在皮肤中特异性表达E1a,并表现出异常的毛发表型,具有明显的区域差异。受影响的毛干短而弯曲,毛囊表现出营养不良或缺乏内根鞘。毛囊间表皮正常,但其对TPA(12-O-十四烷酰佛波醇-13-乙酸酯)急性处理的增生反应明显降低。从这些动物中分离出的原代角质形成细胞对TPA和TGF-β的作用具有部分抗性。转基因小鼠中自发或化学诱导的皮肤肿瘤发生率没有增加。因此,干扰已知负性生长调节蛋白的转基因表达会导致角质形成细胞成熟为高度有组织的结构(如毛囊)受到严重干扰,但不会导致增殖增加和/或肿瘤性增殖。