目的 从人肺腺癌A549细胞系分离出SP亚群细胞,并观察吉非替尼对其生长增殖、细胞周期的影响.方法 经Hoechst33342和碘化丙啶(PI)染色,流式细胞仪分离出具有干细胞特性的SP亚群.用0～40 μmol·L-1浓度的吉非替尼和SP细胞共培养24、48、72 h,采用MTT法检测吉非替尼对细胞增殖的影响;用流式细胞仪检测其对细胞周期分布的影响.结果 SP亚群占A549细胞的1.89%,维拉帕米阻断后减少为0.39%.吉非替尼在0～40 μmol·L-1浓度范围内能明显抑制A549细胞的增殖,并存在浓度和时间依赖性.流式细胞仪检测显示吉非替尼抑制A549 SP细胞增殖是通过阻滞细胞周期使其停留在G0/G1期实现的.结论 A549细胞中存在SP细胞亚群,吉非替尼抑制A549干细胞的增殖是通过阻滞A549 SP细胞于G0/G1期实现的.%Objective To separate the side population (SP) of cancer stem-like cells from lung cancer A549 cells, and to investigate the effect of gefitinib on the proliferation and cell cycle of A549 SP cells. Methods A549 SP cells were isolated by flow cytometry with Hoechst 33342 and PI staining and then were exposed to different concentrations of gefitinib (0-40 μmol · L-1) for 24,48 and 72 hours. Cell proliferation was detected by MTT. Cell cycle distribution was detected by flow cytometry. Results The treatment with verapamil decreased the percentage of SP cells from 1. 89% to 0. 39%. The treatment with gefitinib (0-40 μmol · L-1) significantly inhibited the p roliferation of A549 SP cells in a dose-and time-dependent manner. Flow cytometry showed that gefitinib treatment inhibited the proliferation of A549 SP cells by inducing G0/G1 arrest. Conclusion Lung cancer A549 cells contain SP cells. The gefitinib can inhibit the proliferation of A549 SP cells by causing G0/G1 phase cell cycle arrest.
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