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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 ,可能成为白血病基因治疗的靶基因
In order to investigate the effect of p2 1WAF1 gene on the proliferation of leukemia cells, retroviral vector p2 1WAF1 was constructed and transfected into K562 cells with p2 1WAF1 expression in vitro by FuGENETM6. The G4 18-resistant K5 6 2 cell line was screened and the expression of exogenous p2 1 WAF1 was detected by RT PCR and Western blotting. The cell proliferation was observed by viable cell counting, and the flow cytometry and soft agar colony formation assay were performed simultaneously. The results showed that the expression of P2 1 WAF1 protein and mRNA was detected in K562p21WAF1 cells, and the cell proliferation was significantly slowed down. The number of cells in G0 G1 phase increased and the colony forming ability of cells in soft agar decreased. The above results show that p2 1WAF1 gene can inhibit the proliferation of leukemic cells with p2 1WAF1 negative expression and may be the target gene for gene therapy of leukemia