从更换Gate选取来评估“API技术”中的凋亡细胞存在

目的:研究“API技术”中的凋亡细胞存在并评估“API技术”和常规亚G1峰细胞凋亡检测法的一致性。方法:以0.15μmol· L-1喜树碱诱导MOLT 4细胞凋亡,结果分析从流式细胞术的 FL2 W/FL2 A坐标和 DNA直方图中选取细胞凋亡域Gate来评估“API技术”中细胞凋亡域的存在。并分别比较由“API技术”分析获得的细胞凋亡率和由DNA直方图分析得到的细胞凋亡率之间的差别。结果:采用不同的 Gate选取方法在“API技术”中找到相同的细胞凋亡域,而“API技术”中的细胞凋亡域与前述两种方法找到的细胞凋亡域并不吻合,由“API技术”分析的细胞凋亡率为(24.75±3.45)%,由传统DNA直方图获得的细胞凋亡率为(5.56±1.17)%,两者之间差异存在极显著性(n=8, P<0.01)。结论:“API技术”和亚 G1 峰检测法的细胞凋亡域不一致,“API技术”对细胞周期特异性凋亡的检测结果与亚G1峰检测结果也不一致。

Evaluation Existence of Apoptotic Cells with "API Method" by Changing Gate Selection

Objective: To evaluate existence of apoptotic cells in “API method”, which is regarded as a detection method of cell cycle-specific apoptosis and to weigh coincidence of “API method” and routine sub-G1 method in detecting apoptosis. Methods: Apoptosis was induced in MOLT-4 cells by 0.15 μmol·L -1 camptothecin. Existence of apoptotic cells region in “API method” was evaluated by Gate selection from FL2-W/FL2-A coordinates and DNA histogram in FCM. And percentage of apoptosis was analyzed by “API method” and DNA histogram was compared. Results: With different Gate selection, the same region of apoptotic cells was found. However, the apoptotic cells region in “API method” was different from that analyzed by different Gate selection mentioned before. The percentage of apoptosis analyzed by “API method” and sub-G1 method was respectively (24.75± 3.45 )% and (5.56±1.17)%. There was a significant difference between percentage of apoptosis analyzed by “API method” and sub-G1 method (n=8, P<0.01). Conclusion: Regions of apoptotic cells in “API method” and sub-G1 method were not the same. There is no coincidence in results of apoptotic percentage detected by “API method” and sub-G1 method.