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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 6 2细胞对Vp16的敏感性
Objective To investigate the effect of p2 1WAF1 on the proliferation of leukemia cell line K5 6 2 and its sensitivity to chemotherapeutic drug Vp16. Methods The p2 1WAF1 retrovirus expression vector pLXSN p2 1WAF1 was constructed and pLXSN p2 1WAF1 and empty vector pLXSN neo were transfected into K562 cells without p2 1WAF1 expression by FuGENETM6 in vitro. The expression of G418-resistant K5 6 2 cells The results of RT-PCR and Western blot showed that the gene was expressed in K562 cells after transfection. The effect of p2 1WAF1 on the proliferation and cell cycle of K562 cells was detected by trypan blue staining and flow cytometry. The Vp16 sensitivity of K562 cells transfected with p2 1WAF1 was detected by viable cell counting and MTT assay. Results The growth of K562 cells expressing exogenous p2 1WAF1 was significantly slower than that of control cells. The number of cells in G0 / G1 phase was increased by flow cytometry. Counting by viable cells and MTT assay showed that K5 6 2 p2 1WAF1 cells treated with chemotherapeutic drugs The drug sensitivity of Vp16 was significantly decreased. The IC50 of Vp16 against K562 neo cells was (56.4 ± 6.5) μg / ml, while that of K562 p21WAF1 cells was (131.0 ± 8.7) μg / ml, the difference was significant (P <0.01). Conclusion p2 1WAF1 can inhibit the proliferation of K562 cells, but at the same time it decreases the sensitivity of K562 cells to Vp16