中国生物工程杂志

Objecive: To study the effect of antizyme1 on the cell proliferation and cell apoptosis induced by As2O3 in human leukemia K562 cells. Methods: The base of frameshift was deleted by site-directed mutagenesis assay and the mutation antizyme1 gene was then cloned into vector of pEGFP-N1. Then the recombination DNA was transfected into K562 cells and stable transferctants were isolated after selection with G418. Cell proliferation was checked by MTT assay, and the cell cycle was detected by FCM. The level of cyclin D1 mRNA was measured by RT-PCR assay. After treated with various concentration of As2O3, apoptosis rate of K562pAZ1m was detected by FCM.The level of survivin mRNA was measured by RT-PCR assay also. Results: The mutation antizyme1 gene was successful cloned into pEGFP-N1.　When transfected into K562 cells, the cell proliferation was inhibited and cell cycle was arrested at G0/G1 stage according with down regulation of cyclin D1 expression. After treated with As2O3, apoptosis rate of K562pAZ1m rising is coincident with the decrease of survivin expression. Conclussion: antizyme1 can inhibite proliferation in K562 cells and arrest cell cycle by down regulating cyclin D1 expression. And antizymes also enhance the apoptosis rate induced by As2O3 partly via decreasing survivin expression