Liu J J, Chao J R, Jiang M C, Ng S Y, Yen J J, Yang-Yen H F
Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan, Republic of China.
Mol Cell Biol. 1995 Jul;15(7):3654-63. doi: 10.1128/MCB.15.7.3654.
Ectopic overexpression of v-H-Ras protein in NIH 3T3 cells resulted in cellular transformation and an acceleration of G1 progression of these cells. A shortened G1 phase was found to be associated with an increased level of cyclin D1 but not cyclin E protein. Using an antisense blocking method, reduced synthesis of cyclin D1 in v-H-Ras transformants resulted in a slower G1 progression rate of these cells. Although constitutive overexpression of cyclin D1 in NIH 3T3 cells accelerated G1 progression, cells remained untransformed. Furthermore, inhibition of cyclin D1 synthesis greatly impaired the soft-agar cloning efficiency of v-H-Ras transformants. These results suggest that increased expression of cyclin D1 is necessary but not sufficient for the transforming activity of v-H-Ras. Similar effect on cell cycle progression was also observed in Raf-transformed cells. In addition to cyclin D1, cyclin E protein was found to be elevated in Src transformants. This may account for the further shortening of the G1 phase of these cells. Activation of an additional Ras-independent pathway was suggested to be responsible for the further acceleration of the G1 phase in Src transformants.
v-H-Ras蛋白在NIH 3T3细胞中的异位过表达导致细胞转化,并加速了这些细胞的G1期进程。发现G1期缩短与细胞周期蛋白D1水平升高有关,而与细胞周期蛋白E蛋白无关。使用反义阻断方法,v-H-Ras转化体中细胞周期蛋白D1的合成减少导致这些细胞的G1期进程速率减慢。尽管NIH 3T3细胞中细胞周期蛋白D1的组成型过表达加速了G1期进程,但细胞仍未转化。此外,抑制细胞周期蛋白D1的合成极大地损害了v-H-Ras转化体的软琼脂克隆效率。这些结果表明,细胞周期蛋白D1表达增加对于v-H-Ras的转化活性是必要的,但不是充分的。在Raf转化的细胞中也观察到对细胞周期进程的类似影响。除了细胞周期蛋白D1外,还发现Src转化体中细胞周期蛋白E蛋白升高。这可能解释了这些细胞G1期的进一步缩短。有人提出,激活另一条不依赖Ras的途径是Src转化体中G1期进一步加速的原因。