环孢素A体外诱导HL-60细胞凋亡的研究

A study on the apoptosis in HL-60 cells induced by cyclosporine A

目的：观察环孢素Ａ是否有诱导白血病细胞凋亡的作用。方法：应用细胞形态学检查、二苯胺法ＤＮＡ片段化定量，ＤＮＡ凝胶电泳及流式细胞术等方法观察细胞凋亡。结果：ＣｓＡ５０ｍｇ／Ｌ作用ＨＬ－６０细胞４ｈ，ＤＮＡ片段率显著增高，达（２８．２±５．８）％，而对照组仅为（１２．５±１．７）％（Ｐ＜０．０１，ｎ＝１０）。光镜及电镜检查见细胞核固缩，碎裂，有凋亡小体形成。ＤＮＡ电泳显示典型的ＤＮＡｌａｄｄｅｒ。流式细胞术检测发现ＣｓＡ５０ｍｇ／Ｌ作用ＨＬ－６０细胞６ｈ凋亡细胞率为４９．７％，而空白组仅为９．１％。ＣｓＡ诱导ＨＬ－６０细胞ＤＮＡ片段化呈剂量和时间依赖性。结论：ＣｓＡ在体外能诱导ＨＬ－６０细胞凋亡。

AIM:To investigate if cyclosporine A(CsA) induces the apoptosis in HL-60 cells.METHODS:Apoptosis was measured by the morphology of apoptotic cells,the quantitation of DNA fragmentation with the diphenylamine reaction,the agarose-gel electrophoresis of DNA fragment,and flow cytometric analysis.RESULTS:When the cells were incubated with CsA 50 mg/L for 4h, the percentage of DNA fragmentation increased to (28.2±5 8)% compared to the control group (12.5±1 7)% without CsA ( P <0 01, n =10).Chromatin condensation,nuclear fragmentation and production of apoptotic bodies were seen in the cells treated with CsA utilizing light and electron microscopy.DNA electrophoresis of CsA-treated HL-60 cells showed nuclear fragmentation in oligonucleosomal fragments (DNA ladder).Flow cytometric analysis revealed that the percentage of apoptotic cells was 49.7% and 9.1% after the cells were incubated with or without CsA 50 mg/L respectively.Induction of DNA fragmentation by CsA was both dose-and-time dependent.CONCLUSION:CsA can induce apoptosis in HL-60 cells.