Research For Expressing Of Survivin And Genes Associated With Telomeres In Human Gastric Cancer Cell Line AGS By P21^WAF

Objective To investigate the role of p21WAF gene in the regulation survivin mRNA and telomeres associated genes including hTERT,TRF1,TRF2å’ŒhPOT1 of human gastric cancer cell line AGS.Methods An eukaryotic expression recombinant plasmid of pEGFP-C2-p21 was reconstrcted by clone techology. Liposomal lipofectamine2000 mediates and G418 selected foreign gene p21WAF stable transfection of AGS cells.Report gene pEGFP emitted green fluorescence were observed by fluorescence microscope method, which was estimated the position of p21WAF and the expression efficiency in AGS cell. The cell cycles were analyzed by flow cytometry. The gene expression level of foreign gene p21WAF and the survivin,hTERT,TRF1,TRF2å’ŒhPOT1 gene was detected with real-time PCR.Results Recombinant plasmid pEGFP-C2-p21 was identified correct sequence and direction by PCR, enzyme digestion and DNA sequencing. And also was observed stable expression and localization in cell nuclear by green fluorescent protein. Cell cycle in AGS cells Was blocked in G1 phase.mRNA analysis show the mRNA level of genes including p21,survivin,hTERT,TRF1,TRF2 and hPOT1 in stable transfection p21-AGS cell groups was 324times,1.6%,95%,97%,102% and 99% than that of C2-AGS groups selectively. Conclusion Overexpression of P21WAF down-regulated survivin gene expression at the transcriptional level, may due to regulating cell cycle in cell nuclear. But probably have no effect on genes of hTERT,TRF1,TRF2 and hPOT1.