摘要
本研究旨在探讨三氧化二砷 (arsenictrioxide ,ATO)诱导P2 10 Bcr AblK5 6 2细胞凋亡的发生机制。采用细胞培养、细胞形态观察、流式细胞术 (FCM )测定凋亡细胞和细胞周期 ,应用半定量逆转录 聚合酶链反应 (RT PCR)分析ATO作用不同时间后K5 6 2细胞bcl XL 和bcr abl基因的改变 ,并检测胞浆细胞色素C(cytC)、半胱天冬酶 3(caspase 3)蛋白的表达。结果表明 :2 .5 μmol LATO作用 4 8小时可诱导K5 6 2细胞凋亡 ,凋亡进程中细胞胞浆内cytC蛋白浓度增高 ,caspase 3活性增强 ;RT PCR显示ATO对bcr abl基因表达无明显影响 ,但 12小时可下调bcl XL 基因。结论 :ATO通过激活胞浆线粒体下游的cytC和caspase 3激酶活性而诱导K5 6 2细胞凋亡 ,bcl XL
The aim was to investigate the mechanism of arsenic trioxide (ATO) inducing apoptosis of K56 2 cells that express P210 Bcr-Abl. Apoptosis was analyzed by cell prolif eration assay, morphological changes, DNA-PI staining and cell cycle analysis. ELISA kits were also used to analyze the concentration of cytosolic cytochr ome C and activation of caspase-3. Transcriptional levels of Bcl-X L and Bcr-Abl w ere a ssayed by semiquantitative reverse transcription ploymerase chain reaction (RT- PCR). The results showed that K562 cells were induced to apoptosis after exposur e to 2.5 μmol/L ATO for at least 48 hours, and the cell cycle of K562 cells was arrested at the G 2/M phase. Caspase-3 was activated and there was a cyto solic ac cumulation of cytochrome C. ATO could only reduce the transcriptional level of B c l-X L, but could not down-regulate the Bcr-Abl transcriptional level. In con clusion, ATO can induce K562 cells to apoptosis. The signal pathway mediated by the cyt osolic translocation of mitochondria cytochrome C is one of the mechanisms for ATO inducing apoptosis. And the decrease of Bcl-X L may induce more apoptosis .
出处
《中国实验血液学杂志》
CAS
CSCD
2004年第5期558-562,共5页
Journal of Experimental Hematology
基金
江苏省卫生厅重大项目资助 (编号H2 0 0 2 0 6)
