p21对人胃癌细胞株 AGS的 survivin及端粒相关基因表达水平的调控

IM To investigate the role of P21WAF gene in the regulation survivin mRNA and telomeres associated genes including htert、TRF1、TRF2 and hPOT1 of human colorectal cancer AGS cell line．Methods An eukaryotic expression recombinant plasmid of pEGFP-C2-P21 was reconstrcted by sub-clone techology. Liposomal mediates and G418 selected foreign gene P21WAF stable transfection of lovo cells.Report gene pEGFP emitted green fluorescence were observed by fluorescence microscope method, which was estimated the position of P21WAF in AGS cell. The cell cycles were analyzed by flow cytometry. The gene expression level of foreign geng P21WAF and the survivin、htert、TRF1、TRF2 and hPOT1 gene was detected with real time PCR．Results P21 gene was stable expressed in AGS cell and the sub-cell location was nucleolus. Cell cycle in AGS cells Was blocked in G1/S phase．mRNA analysis show the mRNA level of genes including P21、survivin、htert、TRF1、TRF2 and hPOT1 in stable transfection lovo cell groups was 324times,1.6%,95％,97％,102% and 99% than that of C2-lovo groups selectively. Conclusion Overexpression of P21WAF down-regulated survivin gene expression at the transcriptional level, may due to regulating cell cycle. But probably have no effect on genes of htert、TRF1、TRF2and hPOT1.