白血病细胞系APAF-1cDNA类型与基因表达关系的研究

cDNA types and expression of APAF-1 in leukemic cell lines

目的:探讨白血病细胞系APAF-1cDNA的结构差异及其与基因表达的关系。方法:应用RT-PCR结合测序方法研究白血病细胞系K562和CEM/VLB100APAF-1cDNA类型;应用RT-PCR和Westernblot方法分析两个细胞系APAF-1基因的表达水平;检测caspase-3活性以比较APAF-1基因在两个细胞系的功能差异。结果:测序结果K562和CEM/VLB100APAF-1cDNA分别属于APAF-1XL和APAF-1L型,两个细胞系APAF-1mRNA表达水平无差异。K562细胞中APAF-1蛋白表达水平和caspase-3活性明显低于CEM/VLB100。结论:APAF-1的结构差异影响了APAF-1蛋白表达水平,可能是造成caspase-3活性降低的重要原因。

Objective： To explore whether the structural differences in APAF-1 cDNA types exist and whether such differences affect the function of APAF-1 gene in leukemic cell lines. Methods： APAF-1 cDNA types in leukemic cell lines K562 and CEM/VLB100 were studied by cDNA cloning strategy.The expression of APAF-1 gene was detected by using reverse transcription polymerase chain reaction and Westem blotting. The activities of caspase-3 in these 2 cell lines were detected. Results： APAF-1 cDNA sequences of K562 and CEM/VLB100 belonged to APAF-1XL and APAF-1L,respectively. There was no difference in APAF-1 mRNA level between 2 cell lines, The APAF-1 protein level and the activity of caspase-3 in cell line K562 were significantly lower than those in cell line CEM/VLB100. Conclusion： The structural differences in APAF-1 cDNA types of cell lines K562 and CEM/VLB100 affect the expression of APAF-1 protein,and might be an important factor resulting in the decrease of caspase-3 activity.