Li Xin-Mei, Chen Han-Chun, Liu Xin-Fa, Wu Yan-Hui, Cao Yan-Fei
Molecular Biology Research Center, Xiangya Medical College, Central South University, Changsha, Hunan, 410078, PR China.
Ai Zheng. 2003 Oct;22(10):1047-52.
BACKGROUND & OBJECTIVE: The clinical observation has shown that interferon-alpha(IFN-alpha) is one of the most effective therapeutic agents for the malignancies of hemopoietic system and lymphoma. However, IFN-alpha can only induce about 70-80% of the patients with chronic myeloid leukemia (CML) to get hematological remission. The mechanism by which various CML cases respond differently to IFN-alpha is still unclear.
(1)The effects of IFN-alpha in different concentrations (100, 500, 1,000, 5,000, and 10,000 U/ml) on growth of the two CML cell lines were detected by MTT assay,semisolid colony formation and trypan-blue staining for the cells in liquid culture. (2)The cell apoptosis was examined by flow cytometry(FCM),fluorescence microscopy and gel electrophoresis analysis for DNA fragmentation in 48 hours after IFN-alpha (1,000 U/ml) induction of both KT-1/A3 and K562 cells. (3)The expression levels of bcr/abl chimeric genes were analyzed by relative quantitative RT-PCR at 48 hours after cultivation of both KT-1/A3 and K562 cells with IFN-alpha in 1,000 U/ml.
(1)The growth inhibition of KT-1/A3 cells was dose-dependent in IFN-alpha from the concentration of 100 U/ml to 10,000 U/ml. (2)Having been induced with IFN-alpha in 1000 U/ml for 48 hours, the apoptosis rate of KT-1/A3 cells raised from 3.29% to 11.8% and the expression level of bcr/abl chimeric gene in this cell line declined to 66.7% as compared with those of the control. (3)The growth and apoptosis rate as well as bcr/abl gene expression level of K562 cells were not significantly affected by IFN-alpha.
Different populations of CML cells shows different sensitivity to IFN-alpha.
临床观察表明，α-干扰素（IFN-α）是治疗造血系统恶性肿瘤和淋巴瘤最有效的药物之一。然而，IFN-α仅能使约70-80%的慢性髓性白血病（CML）患者获得血液学缓解。不同CML病例对IFN-α反应不同的机制尚不清楚。
（1）采用MTT法、半固体集落形成试验及台盼蓝染色法，检测不同浓度（100、500、1000、5000和10000 U/ml）IFN-α对两种CML细胞系生长的影响。（2）采用流式细胞术（FCM）、荧光显微镜及DNA片段化凝胶电泳分析，检测IFN-α（1000 U/ml）诱导KT-1/A3和K562细胞48小时后的细胞凋亡情况。（3）采用相对定量RT-PCR法，分析IFN-α（1000 U/ml）培养KT-1/A3和K562细胞48小时后bcr/abl嵌合基因的表达水平。
（1）100 U/ml至10000 U/ml浓度的IFN-α对KT-1/A3细胞的生长抑制呈剂量依赖性。（2）1000 U/ml IFN-α诱导KT-1/A3细胞48小时后，其凋亡率从3.29%升至11.8%，该细胞系中bcr/abl嵌合基因的表达水平降至对照组的66.7%。（3）IFN-α对K562细胞的生长、凋亡率及bcr/abl基因表达水平无明显影响。
不同群体的CML细胞对IFN-α的敏感性不同。
