目的探讨重离子辐照对人肺癌细胞株H1299细胞周期及caspase-3蛋白表达的影响.方法 以不同剂量的12C6+辐照H 1299细胞,培养12、24、48、72 h收获细胞.用流式细胞术和噻唑蓝法检测H1299细胞周期及增殖的变化.caspase-3荧光分析试剂盒检测caspase-3活性的变化.蛋白印迹法和SP法检测辐照后caspase-3蛋白的表达.结果 H1299细胞经12C6+辐照后出现形态学改变；辐照12h细胞出现中期阻滞；4Gy辐照24 h细胞中期阻滞最为明显,与对照组比较差异有统计学意义(P＜0.05),具有剂量依赖性.caspase-3活性随着辐照剂量的增加而增加.12C6+显著上调caspase-3的表达.结论 12C6+辐照能够调控细胞周期,抑制细胞生长.重离子辐照H1299细胞经非p53依赖途径发生凋亡,caspase-3可能在辐照诱导细胞凋亡中发挥重要作用.%Objective To investigate the effect of 12C6+ irradiation on cell cycle and caspase-3 expression in human lung cell line HI299. Methods The changes of cell cycle in HI299 were detected by flow cytometry, the inhibition of cell proliferation was observed by microculture tetrazolium test and the expression of caspase-3 was detected by immunohistochemistry and western blot. Results Showed that after H1299 cells irradiated by 12C6+, the block of G2/M phase began to appear after treatment for 12 h, the most obvious block of G2/M appeared after treatment with 4 Gy for 24 h, which showed dose-dependence. The apoptosis rate tended to increase with the radiation dose increasing. All of the study results involving in Western Blot and immunohistochemistry shown that irradiation of 12C6+ could increase expression of caspase-3 with an obviously statistical significance comparing with control group (P <0.05). ConclusionThe results indicated that irradiation of 12C6+ could induce apoptosis in human lung cell lineH1299 and could significantly inhibit the growth of H1299 cells via caspase-3 pathway activatedby heavy ion irradiation. That caspase-3 may play an important role in the radiation-inducedapoptosis in non-p53-dependent pathway.
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