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

作　　者：李莉[1] 宋振岚[1] 徐东刚[2] 王嘉玺[2] 

机构地区：[1]大连医科大学附属第二医院血液内科,116023 [2]军事医学科学院基础医学研究所

出　　处：《中华血液学杂志》2007年第1期22-26,共5页Chinese Journal of Hematology

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.

关 键 词：肝脂素 细胞凋亡 基因 Wnt13 基因 ATPase3 KG-1细胞 

分 类 号：R686[医药卫生—骨科学]