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目的探讨一氧化氮(NO)是否提高γ射线对L1210细胞的损伤效应及其机制。方法将L1210细胞加入隔离培养器内与3T3细胞共培养,收集经γ射线照射后12、24、48和72h各组L1210细胞,台盼蓝染色观察直接损伤作用,TUNNEL法观察诱导细胞凋亡的情况,流式细胞术检测细胞周期。结果γ射线照射后质粒转染组L1210细胞台盼蓝拒染率下降最快,12h后与空载体组比较差异有统计学意义(P<0·05),加DEVD-CHO后能够提高台盼蓝拒染率,24h后有显著的差异(P<0·05);γ射线照射后质粒转染组细胞凋亡率24h开始明显升高,与空载体组比较差异有统计学意义(P<0·05),加DEVD-CHO后细胞凋亡率下降,24h开始与单独质粒转染组比较差异有统计学意义(P<0·05);射线照射后质粒转染组细胞G1期快速上升,4h与空载体组差异有统计学意义(P<0·05),加DEVD-CHO后G1期上升较单纯质粒转染组慢,4h两组差异有统计学意义(P<0·01)。结论NO可增强γ射线对L1210细胞的直接损伤作用,出现细胞的G1阻滞,促进细胞凋亡。Caspase-3的活化参与上述作用。
Objective To investigate whether nitric oxide (NO) can increase the damage effect of γ-rays on L1210 cells and its mechanism. Methods L1210 cells were cultured in vitro and co-cultured with 3T3 cells. L1210 cells in each group were collected at 12, 24, 48 and 72 hours after γ-ray irradiation. The direct injury was observed by trypan blue staining. The apoptosis induced by TUNNEL method was observed Circumstances, flow cytometry cell cycle. Results The expression of trypan blue was the fastest in L1210 cells transfected with γ-ray and the difference was statistically significant after 12 hours (P <0.05). After adding DEVD-CHO, (P <0.05). The apoptosis rate of plasmid transfection group after 24h irradiation started to increase obviously, which was significantly different from that of blank vector group (P < 0.05). After the addition of DEVD-CHO, the apoptosis rate decreased, and the difference was statistically significant (P <0. 05) between 24h and the plasmid transfection alone group. The G1 phase of the plasmid transfection group increased rapidly (P <0 · 05). The increase of G1 phase after adding DEVD-CHO was slower than that of plasmid transfection group, and the difference was significant at 4h (P <0.01) . Conclusion NO can enhance the direct damage effect of γ-rays on L1210 cells, resulting in G1 arrest and cell apoptosis. Activation of Caspase-3 is involved in these effects.