Stable expression of programmed cell death 4 gene in glioma cell line and its influence on proliferation of tumor cells / 中国肿瘤生物治疗杂志

ABSTRACT

Objective:To establish a glioma cell line U251 stably expressing programmed cell death 4(PDCD4)gene,and to observe the influence of exogenous PDCD4 gene on the proliferation and cell cycle of U251 cells.Methods:Recombinant eukaryotic expression vector pEGFP-PDCD4 was transfected into human glioma cell line U251 by Lipofectamine 2000,and the U251 cells stably expressing PDCD4 were established by G418 selection.Reverse transcription polymerase chain reaction(RT-PCR)and Western blotting were employed to detect the expression of PDCD4 mRNA and protein.Furthermore,cell proliferation and colony forming ability were determined by cell counting and colony formation assay;the cell cycle was detected by FACS.Results:High expression of PDCD4 mRNA and protein was observed in U251 cells transfected with pEGFP-PDCD4,whereas no PDCD4 mRNA and protein expression was detected in the non-transfected and vector-transfected cells.Further,cells transfected with pEGFP-PDCD4 grew more slowly and had lower colony formation rate than cells of the other two control groups(P