甲睾酮诱导人白血病K562细胞凋亡

Induction of apoptosis in human leukemia K562 cells by alpha-anordrin.

作者信息

Lou L G, Xu B

机构信息

Shanghai Institute of Materia Medica, Chinese Academy of Sciences, China.

出版信息

Zhongguo Yao Li Xue Bao. 1996 May;17(3):255-8.

PMID:9812751

Abstract

AIM

To study antitumor action of alpha-anordrin (Ano).

METHODS

Morphological assessment of apoptosis was performed with light microscope and electron microscope. Membrane integrity was determined by trypan blue exclusion method. Endonucleolysis was assessed by agarose gel electrophoresis and flow cytometric methods.

RESULTS

Exposure of exponentially growing K562 cells to Ano 2.5-50 mumol.L-1 for 48 h resulted in growth arrest, Ano 50 mumol.L-1 inhibited the growth of K562 cells by 67%. Cells were mainly blocked to progress through S-phase and arrested at G1 phase. After treatment of K562 cells with Ano, marked morphological changes including condensed chromatin, nuclear fragmentation, and reduction in volume were observed. Agarose gel electrophoresis of DNA from cells treated with Ano for 24-48 h revealed "ladder" pattern, typical features of apoptosis, and near 70% of cells underwent apoptosis as determined by flow cytometry. The S-phase cells were more susceptible to apoptosis. Despite extensive cleavage of DNA and nuclear fragmentation, the cell membrane of Ano-treated cells remained intact, excluding trypan blue. Apoptotic cells were detected as early as 8 h after Ano (50 mumol.L-1) treatment.

CONCLUSION

Ano induces apoptosis in K562 cells.

摘要

目的

研究α-去甲雄三烯醇酮（Ano）的抗肿瘤作用。

方法

采用光学显微镜和电子显微镜对细胞凋亡进行形态学评估。用台盼蓝排斥法测定细胞膜完整性。通过琼脂糖凝胶电泳和流式细胞术评估核酸内切酶活性。

结果

对数生长期的K562细胞暴露于2.5 - 50μmol·L-1的Ano中48小时导致生长停滞，50μmol·L-1的Ano抑制K562细胞生长达67%。细胞主要被阻滞于S期进程并停滞在G1期。用Ano处理K562细胞后，观察到明显的形态学变化，包括染色质浓缩、核碎裂和体积减小。对用Ano处理24 - 48小时的细胞的DNA进行琼脂糖凝胶电泳显示出“梯形”图谱，这是凋亡的典型特征，通过流式细胞术测定近70%的细胞发生凋亡。S期细胞对凋亡更敏感。尽管DNA发生广泛裂解和核碎裂，但用Ano处理的细胞的细胞膜保持完整，台盼蓝不能进入。在用Ano（50μmol·L-1）处理后8小时即可检测到凋亡细胞。