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目的 观察γ-射线诱导HL-60细胞凋亡的时间和剂量效应，以探求辐射诱发肿瘤细胞凋亡分子机制。方法 分别以2.5、5、7.5、10、15Gy γ射线照射HL60细胞，于照后继续培养3、6、9、12、24、27、30h，分别收获细胞。电泳检测DNA梯状带；流式细胞术(FCM)细胞凋亡定量并观察细胞周期变化；Hoechst 33258-PI复染荧光显微镜计数凋亡细胞及已死亡细胞百分率。结果 HL-60细胞受照后死亡方式主要是凋亡，其首先阻滞在G２／M期并由此走向凋亡；DNA梯状带及FCM检测结果发现，细胞凋亡具有时间和剂量依赖性；Hoechst 33258PI染色法检测结果与FCM检测的凋亡细胞随剂量的加大及时间的推移而增多的结果并不呈平行关系。结论 γ-射线诱导HL-60细胞凋亡时间和剂量效应关系的确立为进一步探讨该凋亡途径分子机制奠定了基础。
Objective To observe the time and dose effect induced by γ-rays in HL-60 cells in order to explore the molecular mechanism of radiation-induced apoptosis in tumor cells. Methods HL60 cells were irradiated with 2.5, 5, 7.5, 10 and 15 Gy gamma rays, respectively. The cells were harvested after 3, 6, 9, 12, 24, 27 and 30 hours. The DNA ladder was detected by electrophoresis; the apoptosis of FCM was quantified by flow cytometry (FCM) and the cell cycle was observed; the percentage of apoptotic cells and dead cells were counted by Hoechst 33258-PI staining. Results The apoptotic mode of HL-60 cells was mainly apoptosis after irradiation, which firstly blocked in G2 / M phase and then led to apoptosis. DNA ladder and FCM showed that apoptosis was time-and dose-dependent ; Hoechst 33258PI staining results and FCM detection of apoptotic cells with increasing dose and the passage of time and the results of the increase is not parallel. Conclusion The establishment of the relationship between γ-ray-induced HL-60 cell apoptosis time and dose-response laid the foundation for further exploration of the molecular mechanism of apoptosis.