摘要
目的：了解柔红霉素（ＤＮＲ）体外作用于ＨＬ－６０ 细胞时产生凋亡的规律及一些相关蛋白表达的变化。方法：应用光镜、ＤＮＡ电泳及ＦＣＭ检测ＨＬ－６０细胞发生的凋亡模式；用ＩＣ、ＦＣＭ观察相关蛋白的变化。结果：当ＤＮＲ浓度为０．２μＭ～２μＭ时，ＨＬ－６０细胞发生的凋亡率随浓度的增加与作用时间的延长而升高，可见典型的凋亡细胞及明显的ＤＮＡ梯度条带。１μＭ ＤＮＲ作用２ｈ时，ｂｃｌ－２与ｃ－ｍｙｃ的荧光强度指数（ＦＩ）降低，ｂａｘ、ｃａｓｐａｓｅ－３ 的 ＦＩ 升高；而作用５ｈ时，ｂａｘ、ｃａｓｐａｓｅ－３的ＦＩ值反而降低。结论：一定浓度的ＤＮＲ在体外可诱导ＨＬ－６０细胞凋亡，它可抑制ｂｃｌ－２与ｃ－ｍｙｃ蛋白的表达，而激活ｂａｘ、ｃａｓｐａｓｅ－３蛋白的表达。
Objective: To learn about the apoptosis role of HL-60 cells induced by DNR in vitro and the expression change of some protein. Methods: This study observed the apoptosis of HL-60 by optic microscope ,flow cytometry (FCM) and DNA electrophoresis and detected changes of various apoptosis-associated proteins expression with immunocytochemistry (IC) and FCM assays. Results: When treated with 0.2μM～2μM DNR , the percentage of apoptotic HL-60 cells increased with the increase of dose and the extension of time, and there showed the typical apoptotic cells and the appearance of apoptotic DNA ladder . It was shown that after HL-60 cells were treated with 1uM DNR for 2h, the fluorescence intensity index(FI) of both bcl-2 and c-myc decreased ,bax and caspase-3 increased; at 5h, the FI of bax and caspase-3 decreased on the contrary. Conclusion: It is suggested from the results that at the certain concentration, DNR can induce the apoptosis of HL-60 in vitro , and it can suppress the expression of bcl-2,c-myc and activate bax and caspase-3.
出处
《重庆医科大学学报》
CAS
CSCD
2002年第1期59-61,共3页
Journal of Chongqing Medical University
