[Exogenous antizyme 1 gene transfection inhibits proliferation and promotes apoptosis of human neuroblastoma SH-SY5Y cells in vitro].
OBJECTIVE
To study the effect of antizyme 1 (ZA1) gene transfection on the cell proliferation, cell cycle and apoptosis of human neuroblastoma SH-SY5Y cells in vitro.


METHODS
The recombinant eukaryotic expression vector pAZ1m was constructed by cloning mutant AZ1 gene into the vector pEGFP-N1, and subsequently transfected in SH-SY5Y cells. The transfected cell proliferation was examined using MTT assay, and the changes in cell cycle and apoptosis were assayed using flow cytometry analysis. RT-PCR was performed to measure cyclin D1 and caspase-3 mRNA expressions, Western blotting carried out to examine cyclin D1 protein expression, and the changes in caspase-3 activity were detected using a caspase-3 detection kit.


RESULTS
AZ1 gene transfection significantly inhibited the proliferation of SH-SY5Y cells, causing cell cycle arrest at G0/G1 stage and down-regulated cyclin D1 and up-regulated caspase-3 expressions. Obviously increased caspase-3 activity was also observed in the transfected cells.


CONCLUSIONS
Exogenous AZ1 gene transfection can inhibit the proliferation and cause cell cycle arrest of SH-SY5Y cells by down-regulating cyclin D1 expression. Up-regulated caspase-3 expression resulting from AZ1 gene transfection may induce apoptosis of the neuroblastoma cells.
Jiang
Li
L
Institute of Genetic Engineering, Southern Medical University, Guangzhou 510515, China. jiangli27556@126.com
Ma
Wen-li
WL
Li
Jin
J
Peng
Yi-fei
YF
Xu
Bing
B
Zheng
Wen-ling
WL
D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D019938:Cyclin D1 / Q000378:metabolism; D015536:Down-Regulation; D005822:Genetic Vectors; D006801:Humans; D009447:Neuroblastoma / Q000378:metabolism*; D011506:Proteins / Q000235:genetics*; D014162:Transfection; D015854:Up-Regulation
D011506:Proteins; C031017:ornithine decarboxylase antizyme; D019938:Cyclin D1; D053148:Caspase 3