Chiang Y, Rizzino A, Sibenaller Z A, Wold M S, Vishwanatha J K
Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha 68198-4525, USA.
Mol Cell Biochem. 1999 Sep;199(1-2):139-47. doi: 10.1023/a:1006942128672.
The protein-tyrosine kinase substrate annexin II is a growth regulated gene whose expression is increased in several human cancers. While the precise function of this protein is not understood, annexin II is proposed to be involved in multiple physiological activities, including DNA synthesis and cell proliferation. Targeted disruption of the annexin II gene affects calcium signaling, tyrosine phosphorylation and apoptosis, indicating the important physiological role of this protein. We used a transient co-transfection assay to regulate annexin II expression in human HeLa, 293 and 293T cells, and measured the effects of annexin II down regulation on DNA synthesis and proliferation. Transfection of cells with an antisense annexin II vector results in inhibition of cell division and proliferation, with concomitant reduction in annexin II message and protein levels. Cellular DNA synthesis is significantly reduced in antisense transfected cells. Replication extracts made from antisense transfected cells have significantly reduced efficiency to support SV40 in vitro DNA replication, while the extracts made from sense transfected cells are fully capable of replication. Our results indicate an important role of annexin II in cellular DNA synthesis and cell proliferation.
蛋白质酪氨酸激酶底物膜联蛋白II是一种受生长调节的基因，其表达在多种人类癌症中增加。虽然该蛋白质的确切功能尚不清楚，但膜联蛋白II被认为参与多种生理活动，包括DNA合成和细胞增殖。膜联蛋白II基因的靶向破坏会影响钙信号传导、酪氨酸磷酸化和细胞凋亡，表明该蛋白质具有重要的生理作用。我们使用瞬时共转染试验来调节人HeLa、293和293T细胞中膜联蛋白II的表达，并测量膜联蛋白II下调对DNA合成和增殖的影响。用反义膜联蛋白II载体转染细胞会导致细胞分裂和增殖受到抑制，同时膜联蛋白II的信使核糖核酸和蛋白质水平降低。反义转染细胞中的细胞DNA合成显著减少。从反义转染细胞制备的复制提取物在体外支持SV40 DNA复制的效率显著降低，而从正义转染细胞制备的提取物完全能够进行复制。我们的结果表明膜联蛋白II在细胞DNA合成和细胞增殖中起重要作用。
