内质网相关的凋亡途径在高三尖杉酯碱诱导mutz1细胞凋亡过程中的作用.docVIP

内质网相关的凋亡途径在高三尖杉酯碱诱导MUTZ1细胞凋亡过程中的作用 nbsp;nbsp;nbsp;nbsp;nbsp;nbsp;nbsp;nbsp;nbsp;nbsp;nbsp; 作者：胡洁　何冬花　高良　杨春虎 蔡真 【摘要】nbsp; 　　本研究探讨高三尖杉酯碱 ( Homoharringtonine, HHT )诱导人MDS细胞株MUTZ-1细胞凋亡的作用机制。 以HHT处理骨髓增生异常综合征细胞株MUTZ-1（MDS-RAEB）, 用流式细胞术检测细胞凋亡率，用激光共聚焦显微镜检测细胞内钙离子浓度和线粒体膜电位变化，应用RT-PCR检测HHT处理前后与内质网应激有关的基因（CHOP,BIP,XBP1）的表达情况。 结果表明： 0.05 μg/ml HHT作用MUTZ-1细胞的凋亡率随着时间的延长而增加，其差异具有统计学意义（Plt;0.01）；激光共聚焦结果显示， 0.05 μg/ml HHT作用后4小时胞浆内Ca2+浓度增高，12小时达高峰，其后开始下降； HHT作用后线粒体膜电位持续下降； CHOP以及 BIP,XBP1基因表达增强，12小时达到高峰，其后开始下降。 结论: HHT可以诱导MDS细胞株MUTZ-1凋亡，内质网相关的凋亡途径在其中发挥了重要的作用。 【关键词】nbsp; 高三尖杉酯碱； MDS细胞株MUTZ-1 ； 细胞凋亡； 内质网应激 　　Role of Endoplasmic Reticulum Pathway in Apoptosis of MDS Cells Induced by Homoharringtonine 　　AbstractThe study was aimed to investigate the apoptotic mechanism of homoharringtonine (HHT)nbsp; on MDS cell line ( MUTZ-1 cells ).nbsp; The apoptosis of MUTZ-1 cells induced by HHTnbsp; was measured by flow cytometry. The concentration of cystosolic calcium was measured with the mitochondrial membrane potential by confocal . The mRNA expression level of CHOP, BIP and XBP1 were detected by semi-quantitative RT-PCR.nbsp; The results showed that after exposure of MUTZ-1 to HHT for different hours , the percentage of apoptotic MUTZ-1 cells increased in a time dependent manner. Typical apoptotic cells and release of Ca2+ from the cytosolic Ca2+ storage and the loss ofnbsp; mitochondrial membrane potential were observed.nbsp; RT-PCR analysis revealed that mRNAs for endoplasmic reticulum (ER) stress-associated proapoptotic factor CHOP andnbsp; ER chaperone BIP,nbsp; XBP1nbsp; markedly increased at 4 hours afternbsp;nbsp; treatinent with 0.05 μg/ml HHTnbsp; and reached the top level at 12 hours, and then decreased.nbsp; It is concludednbsp; that HHT not only inhibits MUTZ-1 growth but also induces MUTZ-1 apoptosis. The HHT-induced MUTZ-1 cell death is likely mediated by the ER stress pathway. 　　Key wordshomoharringtonine;nbsp;nbsp; MUTZ-1 cell;nbsp; apoptosi