转染p21^(WAF1)基因对K562白血病细胞系增殖的抑制作用  

p21^(WAF1)Gene Transfection Inhibits Proliferation of Leukemia Cell Line K562

作　　者：杨慧[1] 王申五[2] 殷慧君[1] 

机构地区：[1]北京大学人民医院儿科,北京100044 [2]北京大学人民医院中心实验室,北京100044

出　　处：《中国实验血液学杂志》2001年第2期115-118,共4页Journal of Experimental Hematology

摘　　要：为了探讨p2 1WAF1基因对白血病细胞增殖的影响 ,构建了p2 1WAF1的逆转录病毒表达载体 ,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G4 18抗性K5 6 2细胞系 ,用RT PCR和Western印迹检测到外源 p2 1WAF1的表达 ,用活细胞计数观察细胞增殖情况 ,同时进行流式细胞术和软琼脂集落形成实验。结果表明 ,K5 6 2 p2 1WAF1中可检测到P2 1WAF1蛋白及mRNA表达 ,细胞增殖明显减慢 ,G0 G1期细胞数增多 ,细胞在软琼脂中的集落形成能力下降。以上结果表明 ,p2 1WAF1基因可抑制 p2 1WAF1表达阴性的白血病细胞增殖 。To explore the functional role of p21 WAF1 gene on the proliferation of leukemia cell line K562, a p21 WAF1 retroviral expression vector was constructed. Mediated by FuGENE TM 6, p21 WAF1 was transfected into leukemia cell line K562, which was without p21 WAF1 expression. After selected in G418, K562 cell clones that expressed p21 WAF1 stabaly were isolated and named K562 p21 WAF1 . The ectopic expression of p21 WAF1 mRNA and protein in K562 cells was identified by RT PCR and Western Blot. The cell proliferaton was tested in liquid and soft agar culture after transfection. The cell cycle was tested by FCM. 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 clones cultured in liquid media as well as soft agar (P<0.01). The cell number in G 0/G 1 phase was remarkably increased. The findings showed that p21 WAF1 can inhibit the proliferation of leukemia cells, and it could be a potential target gene for leukemia gene therapy.

关 键 词：P21WAF1 K562白血病细胞系 细胞增殖抑制 白血病 基因治疗 

分 类 号：R733.7[医药卫生—肿瘤] R730.54[医药卫生—临床医学]