p21^(WAF1)抑制K562细胞生长并降低其对足叶乙甙的敏感性  

P21^(WAF1)inhibits the growth of leukemia cell line K562 and decreases its sensitivity to Vp16

作　　者：杨慧[1] 王申五[1] 殷慧君[1] 

机构地区：[1]北京大学人民医院中心实验室,100044

出　　处：《中华血液学杂志》2002年第1期19-22,共4页Chinese Journal of Hematology

摘　　要：目的　探讨p2 1WAF1对白血病细胞系K5 6 2增殖及其对化疗药物足叶乙甙 (Vp16 )敏感性的影响。方法　构建p2 1WAF1逆转录病毒表达载体pLXSN p2 1WAF1,将pLXSN p2 1WAF1和空载体pLXSN neo通过FuGENETM6介导体外转染p2 1WAF1表达缺如的K5 6 2细胞 ,经筛选得到G418抗性K5 6 2细胞株 ,用RT PCR和Westernblot证明该基因在转染后的K5 6 2细胞中有表达 ,用锥虫蓝染色法和流式细胞术检测p2 1WAF1对K5 6 2细胞增殖和细胞周期的影响 ,用活细胞计数法和MTT法检测转染p2 1WAF1的K5 6 2细胞对Vp16敏感性变化。结果　表达外源性p2 1WAF1的K5 6 2细胞生长明显慢于对照组细胞 ,流式细胞术检测显示G0 /G1期细胞增多 ,活细胞计数法和MTT法显示K5 6 2 p2 1WAF1细胞对化疗药物Vp16的药物敏感性明显降低 ,Vp16对K5 6 2 neo细胞的IC50 值为 (5 6 .4± 6 .5 ) μg/ml,而对K5 6 2 p2 1WAF1细胞的IC50 值为(131.0± 8.7) μg/ml,两者相比差异有显著性 (P <0 .0 1)。 结论　p2 1WAF1能抑制K5 6 2细胞增殖 ,但同时降低了K5 6Objective To explore the effect of p21 WAF1 on the proliferation and the sensitivity to Vp16 of leukemia cell line K562. Methods A p21 WAF1 retroviral expression vector pLXSN p21 WAF1 was constructed by FuGENE TM 6, pLXSN p21 WAF1 and pLXSN neo, and transfected into p21 WAF1 defect leukemia cell line K562. After selected with G418,K562 p21 WAF1 cell clones that stably expressed p21 WAF1 were isolated. The ectopic expressions of p21 WAF1 mRNA and protein in K562 p21 WAF1 were identified by RT PCR and Western b1ot. The cell growth rate was tested by trypan blue dye, the cell cycle by FCM and the sensitivity to Vpl6 by cell count and MTT assay. Results The expression of p21 WAF1 protein and mRNA could be detected in K562 p21 WAF1 cells. A strong inhibition of cell proliferation was observed in K562 p21 WAF1 cell as compared with that of the control. The cell number in G 0/G 1 phase was remarkably increased. The sensitivity to Vpl6 decreased, the IC 50 of K562 neo cells was (56.4±6 5) μg/ml, and that of K562 p21 WAF1 cells was (131.0±8.7) μg/ml(P<0.01). Conclusion p21 WAF1 can inhibit the proliferation of leukemia cell and decrease its sensitivity to Vp16.

关 键 词：P21^WAF1基因 K562细胞系 增殖抑制 药物耐受性 白血病 

分 类 号：R733.7[医药卫生—肿瘤]