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应用mRNA差异显示法检测肝脂素诱导的KG-1细胞凋亡相关基因表达的改变

Identification of Heplipin inducing apoptosis-related genes in KG-1 human leukemia cells by differential display RT-PCR
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摘要 目的观察肝脂素(heplipin)对KG-1细胞的凋亡诱导作用。比较肝脂素诱导KG-1细胞凋亡前后的基因表达谱,寻找与白血病细胞凋亡相关的基因。方法应用DNA倍体分析和DNA凝胶电泳的方法,证实肝脂素可诱导KG-1细胞凋亡。采用差异显示逆转录聚合酶链反应(DDRT—PCR)的方法寻找肝脂素诱导前与诱导16、20h后的基因表达差异片段,并对其进行克隆分析。结果250μg/ml肝脂素作用KG-1细胞16、20h细胞凋亡率分别为13.5%和30.4%,显著高于未经肝脂素诱导组(1.5%)。肝脂素诱导KG-1细胞凋亡前后存在明显的基因表达差异,Wnt13和ATPase 3基因是肝脂素诱导KG-1细胞凋亡相关下调基因。结论肝脂素诱导KG-1细胞凋亡,其机制是一个多基因参与的过程。肝脂素诱导KG-1细胞凋亡与Wnt13和ATPase 3基因有关。Wnt13基因在多种肿瘤细胞中高表达,在白血病细胞系中被检测到尚属首次。 Objective To investigate whether Heplipin can induce KG-1 cell apoptosis and explore apoptosis related differentially expressed genes in KG-1 leukemia cell before and after Heplipin induction. Methods DNA distribution and DNA electrophoresis were used to prove that Heplipin can induce KG-1 cell apoptosis. The differential display reverse transcription-polymerase chain reaction (DDRT-PCR) was adopted to screen differentially expressed genes before and after Heplipin induction of KG-1 cells for 16 hours and 20 hours. The differentially expressed genes were cloned and analyzed. Results Heplipin could induce KG-1 cell apoptosis. There were differentially expressed genes in KG-1 cells before and after induction. Wnt13 and ATPase 3 were apoptosis related differentially downregulated genes after Heplipin induction. Conclusion Heplipin can induce KG-1 cell apoptosis. Heplipin induced KG-1 cell apoptosis is related with Wnt13 and ATPase3 (PSMC3). It is the first report that Wnt13 was detected in leukemia cell line.
出处 《中华血液学杂志》 CAS CSCD 北大核心 2007年第1期22-26,共5页 Chinese Journal of Hematology
关键词 肝脂素 细胞凋亡 基因 Wnt13 基因 ATPase3 KG-1细胞 Heplipin Apoptosis Gene,Wnt13 Gene, ATPase3 KG-1 cells
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