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Abstract In order to make sure whether expression of cyclin G2 gene promotes or inhibits cell proliferation in vitro, and to investigate its mechanism. we have cloned the whole length of cyclin G2 cDNA by reverse transcription - polymerase chain reaction (RT-PCR ) , and inserted it into pIRESneo eukaryotic expression vector, testified pIRES-G2 recombinant plasmid constructed successfully. The construct was then introduced into HeLa and CV-1 cells in order to observe the effect of cyclin G2 transgene expression on the colony forming capacity, and protein expression of p21WAF1 in transfectant cells was examined by cellular immunochemical staining. The results showed that colony-forming efficiency of the cells transfected with pIRES-G2 construct was much lower compared with that with the control parental vector pIRESneo. Furthermore, pIRES-G2 transfected HeLa cells showed a senescent morphology. The experimental group of CV-1 cells could hardly form any detectable colony, while the corresponding empty vector group showed large patches of colony. More HeLa cells transfected with pIRES-G2 plasmid DNA showed positive p21WAF1 staining and labeling intensity increased significantly compared with its corresponding transfectant cells with the empty control vector (p<0.01). So we conclude that ectopic over expression of cyclin G2 inhibit in vitro cell proliferation of cancer and normal cells and it may negatively regulate the cell cycle by upregulating p21WAF1 expression .
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Received: 23 February 2006
Published: 25 June 2006
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