转染P21 WAF1基因对BT-325细胞端粒酶表达的影响

Effect on telomerase activity of BT-325 cells by exogenous p21 WAF1 expression

目的 :观察外源性p2 1WAF1基因对BT - 32 5细胞的作用 ,探讨p2 1WAF1表达水平的改变对细胞端粒酶活性的影响。方法 :经脂质体介导的方法将PCEP -WAF1质粒导入BT - 32 5细胞中 ,细胞克隆转移扩大培养后 ,采用TRAP -PCR -ELISA、TUNEL和电镜等技术检测外源性p2 1WAF1基因对BT - 32 5细胞端粒酶活性和细胞凋亡的作用。结果 :较对照组相比 ,转染外源性p2 1WAF1基因的BT - 32 5细胞端粒酶表达水平显著下降 ,伴有显著细胞凋亡现象。结论 :外源性p2 1WAF1基因的表达可促进BT - 32 5细胞端粒酶活性降低 ,诱发细胞显著凋亡。提示端粒酶的活性表达可能与细胞周期调控存在密切联系。

Objective:To observe the effect of exogenous p21 WAF1 expression on BT-325 cells and to investigate p21 WAF1 expression on cellular apoptosis and telomerase activity.Methods:[WT5BZ]PCEP-WAF1(p21 WAF1 cDNA gene eucaryotic expression vector) was introduced into BT-325 cells by using lipofectin mediation;TUNEL and TRAP in combination of ELISA were used to determine BT-325/PCEP-WAF1 apoptosis and telomerase activity after cloning. Results:Compared with control,BT-325/PCEP-WAF1 presented a significant drop in telomerase activity expression,accompanied by remarkable apoptosis.Exogenous p21 WAF1 over-expression could inhibit cell proliferation.Conclusion:Exogenous p21 WAF1 expression promotes apoptosis of BT-325 cells and reduces telomerase activity and can inhibit cell proliferation,resulting in a delay cell cycle.