摘要
本研究探讨阿霉素体外作用于K562细胞的细胞效应及癌基因Gfi-1和相关凋亡基因表达变化的机制。用不同浓度的阿霉素作用于K562细胞24小时,然后应用DNA电泳和流式细胞术检测K562细胞凋亡;用RT-PCR、流式细胞术检测Gfi-1、Bcl-2、bax基因和蛋白表达的变化。结果表明:当阿霉素浓度在0、0.5、2.0mg/L作用K562细胞24小时,细胞凋亡增加,可见典型的DNA断裂电泳条带;同时,当ADM浓度自0.5增为2.0mg/L时,Gfi-1表达减少,bax表达增加;Bcl-2表达在ADM0.5-2.0mg/L之间变化不明显,mRNA及蛋白水平均无统计学差异;当阿霉素浓度大于2.0mg/L时,细胞凋亡率并不增加,而是下降,且出现细胞坏死。结论:一定浓度阿霉素能诱导K562细胞凋亡,细胞凋亡率表达的变化与阿霉素浓度呈一定的量效依赖关系,阿霉素诱导K562细胞凋亡可能与抑制Gfi-1基因的表达和激活bax基因的表达有一定的关系。
The study was purposed to explore the effect of adriamycin （ADM） on K562 cells in vitro and the mechanism of expression changes of relevant apoptotic genes and oncogene Gfi-1. The apoptosis was assayed by flow cytometry （FCM） and the DNA electrophoresis; the expression changes of Gfi-1 、Bcl-2、bax mRNA and protein were detected by RT-PCR and FCM after K562 cells were treated with different concentrations of ADM for 24 hours. The results showed that when K562 cells were treated with 0 - 2.0 mg/L ADM for 24 hours, the typical apoptotic DNA electrophoresis band of K562 cells were observed with the dose increasing, When concentration of ADM was 0, 5 and 2, 0 mg/L, the expression of Gfi-1 decreased and the expression of bax increased; when concentration of ADM was 0, 5 -2, 0 mg/ L, the expression of Bcl-2 was not found to be significantly changed, the levels of Bcl-2 mRNA and protein were of no statistical difference, When dose of ADM was higher than 2, 0 mg/L, the percentage of apoptotic K562 cells decreased with cell necrosis. It is concluded that at certain range of concentration, apoptosis or necrosis of K562 cells can be induced by ADM, the percentage of apoptosis, the changes of expression of Bcl-2, bax and Gfi-1 depend on the dose of ADM, The mechanism of apoptosis in K562 ceils induced by ADM may be related to suppression of Gfi-1 oncogene and activation of expression of bax gene.
出处
《中国实验血液学杂志》
CAS
CSCD
2007年第2期278-282,共5页
Journal of Experimental Hematology
